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TW202525846A - Anti-il-13 multispecific antibody constructs and uses thereof - Google Patents

Anti-il-13 multispecific antibody constructs and uses thereofDownload PDF

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TW202525846A
TW202525846ATW113131850ATW113131850ATW202525846ATW 202525846 ATW202525846 ATW 202525846ATW 113131850 ATW113131850 ATW 113131850ATW 113131850 ATW113131850 ATW 113131850ATW 202525846 ATW202525846 ATW 202525846A
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amino acid
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acid sequence
substitutions
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衍 鄭
捷 湯
雁 王
銳 張
徐茗慧
文彥 沈
唐丹明
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美商普羅特歐拉吉克適美國公司
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Abstract

The present invention relates to multispecific constructs that specifically bind to IL-13 and a second target antigen (e.g., TSLP, IL-17A, IL-17F, TNF-[alpha], and IFN-[gamma]). Also provided are antibody constructs that specifically bind to IL-13. Compositions, kits, methods of use (e.g., treating inflammatory disease), and methods of making thereof are also provided.

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Translated fromChinese
抗IL-13多特異性抗體構築體及其用途Anti-IL-13 multispecific antibody constructs and their uses

相關申請案之交互參考Cross-reference to related applications

本申請案主張於2023年8月25日提申之美國臨時申請號63/578,946的權益及優先權，其內容通過引用整體併入本文。This application claims the benefit of and priority to U.S. Provisional Application No. 63/578,946, filed on August 25, 2023, the contents of which are incorporated herein by reference in their entirety.

電子序列表之參考References to Electronic Sequence Listings

電子序列表之內容(253272000140seqlist.xml；大小：239,418位元；及創建日期：2024年8月19日)以引用方式整體併入本文。The contents of the electronic sequence listing (253272000140seqlist.xml; size: 239,418 bytes; and creation date: August 19, 2024) are incorporated herein by reference in their entirety.

本發明係有關特異性地結合至IL-13及第二標靶(例如，TSLP)的多特異性構築體。本文進一步提供分離的抗IL-13抗體構築體、含有該多特異性構築體的醫藥組成物、使用該多特異性構築體治療發炎性疾病的方法及含有該多特異性構築體的套組。The present invention relates to multispecific constructs that specifically bind to IL-13 and a second target (e.g., TSLP). Further provided herein are isolated anti-IL-13 antibody constructs, pharmaceutical compositions containing the multispecific constructs, methods of using the multispecific constructs to treat inflammatory diseases, and kits containing the multispecific constructs.

許多異常細胞與組織以及許多疾病皆表現出獨特的抗原，該些抗原可用於進行免疫細胞媒介的清除，包括發炎性疾病，諸如自體免疫疾病或氣喘。此等發炎性疾病可各自表現出一或多種不同的標靶抗原或相同標靶抗原之一或多種不同的表位。舉例而言，一些抗原在發炎的組織中被過度表現、誘變或選擇性地誘變。因此，全身性或局部性發炎疾病中存在的抗體標靶特異性抗可用作治療劑。Many abnormal cells and tissues, as well as many diseases, express unique antigens that can be exploited for immune cell-mediated clearance, including inflammatory diseases such as autoimmune diseases and asthma. Each of these inflammatory diseases may express one or more distinct target antigens or one or more distinct epitopes of the same target antigen. For example, some antigens are overexpressed, mutated, or selectively mutated in inflamed tissues. Therefore, antibodies specific for the targets of systemic or localized inflammatory diseases may be useful therapeutic agents.

介白素(IL)-13為一種多效性細胞激素，常與氣喘及異位性皮膚炎相關聯。IL-13結合至介白素13受體-α1 (IL-13Rα1)與介白素4受體-α (IL-4Rα)組成的高親和力異質複合體。IL-13亦結合至介白素13受體-α2 (IL-13Rα2)，然而IL-13Rα2已被確定為與IL-13Rα1參與的傳訊級聯不同的誘餌受體。IL-13主要由T輔助2型細胞(Th2細胞)及2型先天類淋巴細胞(ILC2)分泌。ILC2在過敏發炎期間產生IL-5及IL-13，並橋接先天與後天免疫反應。ILC2活性促進T輔助2型細胞(Th2)反應，且ILC2細胞與Th2細胞的共同作用亦與發炎性疾病(諸如過敏及自體免疫病症)相關聯。Interleukin (IL)-13 is a pleiotropic cytokine often associated with asthma and atopic dermatitis. IL-13 binds to a high-affinity heterocomplex composed of interleukin 13 receptor-α1 (IL-13Rα1) and interleukin 4 receptor-α (IL-4Rα). IL-13 also binds to interleukin 13 receptor-α2 (IL-13Rα2), although IL-13Rα2 has been identified as a decoy receptor that participates in a different signaling cascade than IL-13Rα1. IL-13 is primarily secreted by T helper type 2 cells (Th2 cells) and type 2 innate lymphoid cells (ILC2s). ILC2s produce IL-5 and IL-13 during allergic inflammation and bridge innate and adaptive immune responses. ILC2 activity promotes T helper type 2 (Th2) responses, and the combined action of ILC2 and Th2 cells is also associated with inflammatory diseases such as allergies and autoimmune disorders.

抗IL-13抗體臨床試驗，包括失敗的曲羅蘆單抗(tralokinumab)的最新結果(AdbryTM；參見例如Brightling等人之Lancet Respir Med. 2015, 3(9):692-701)及用於治療嚴重氣喘的來瑞珠單抗(lebrikizumab)的混合結果(參見例如Hanania等人之Lancet Respir Med. 2016, 4(10):781-796)證實，需要其他抗IL-13抗體來治療IL-13相關疾病，特別是發炎性疾病。特別地，來瑞珠單抗的ACOUSTICS臨床試驗由於73%的患者出現不良事件而提前終止(參見Szefler等人之Clin Transl Allergy 2022, 12(7):212176)，此進一步表明需要更安全及更有效的治療選項。Clinical trials of anti-IL-13 antibodies, including the recent results of the failed tralokinumab (Adbry™ ; see, e.g., Brightling et al., Lancet Respir Med. 2015, 3(9):692-701) and the mixed results of lebrikizumab for the treatment of severe asthma (see, e.g., Hanania et al., Lancet Respir Med. 2016, 4(10):781-796), demonstrate the need for additional anti-IL-13 antibodies to treat IL-13-related diseases, particularly inflammatory diseases. In particular, the ACOUSTICS clinical trial of relizumab was terminated prematurely due to adverse events in 73% of patients (see Szefler et al. Clin Transl Allergy 2022, 12(7):212176), further demonstrating the need for safer and more effective treatment options.

本發明提供特異性地結合至IL-13及第二標靶(例如，TSLP)的多特異性構築體、包含該多特異性構築體的醫藥組成物及使用其多特異性構築體治療發炎性疾病的方法。進一步提供分離的抗IL-13抗體構築體。The present invention provides multispecific constructs that specifically bind to IL-13 and a second target (e.g., TSLP), pharmaceutical compositions comprising the multispecific constructs, and methods of using the multispecific constructs to treat inflammatory diseases. Furthermore, isolated anti-IL-13 antibody constructs are provided.

在本發明之一態樣中，提供一種多特異性構築體，其包含：(1)特異性地結合至介白素-13 (IL-13)的第一抗體部分，以及(2)特異性地結合至第二抗原的第二抗體部分。在一些實施例中，該第二抗原為由免疫細胞產生的蛋白質。在一些實施例中，該第二抗原為胸腺基質淋巴球生成素(TSLP)。In one aspect of the present invention, a multispecific construct is provided, comprising: (1) a first antibody portion that specifically binds to interleukin-13 (IL-13), and (2) a second antibody portion that specifically binds to a second antigen. In some embodiments, the second antigen is a protein produced by an immune cell. In some embodiments, the second antigen is thymic stromal lymphopoietin (TSLP).

在根據上述多特異性構築體之任一者的一些實施例中，該第一抗體部分包含重鏈可變區(VH1)及輕鏈可變區(VL1)，其中：該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多3個胺基酸變異的變體，(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多3個胺基酸變異的變體，以及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多3個胺基酸變異的變體，且該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1或其含有至多3個胺基酸變異的變體，(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多3個胺基酸變異的變體，以及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多3個胺基酸變異的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80%序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80%序列同一性的變體。在一些實施例中，該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1，(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2，以及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3，且該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1，(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。In some embodiments according to any of the above multispecific constructs, the first antibody portion comprises a heavy chain variable region (VH1) and a light chain variable region (VL1), wherein: the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to 3 amino acid variations, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to 3 amino acid variations, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to 3 amino acid variations, and the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to 3 amino acid variations, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to 3 amino acid variations, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to 3 amino acid variations. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65 or a variant thereof having at least about 80% sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69 or a variant thereof having at least about 80% sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, and the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70, (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69.

在根據上述多特異性構築體之任一者的一些實施例中，該第一抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、sdAb及scFv。In some embodiments according to any of the above multispecific constructs, the first antibody portion is selected from the group consisting of: a full-length antibody, Fab, Fab', F(ab')2 , sdAb, and scFv.

在根據上述多特異性構築體之任一者的一些實施例中，該第一抗體部分為scFv (「抗IL-13 scFv」)。在一些實施例中，該抗IL-13 scFv含有SEQ ID NO:108之胺基酸序列。In some embodiments according to any of the above multispecific constructs, the first antibody portion is a scFv ("anti-IL-13 scFv"). In some embodiments, the anti-IL-13 scFv comprises the amino acid sequence of SEQ ID NO: 108.

在根據上述多特異性構築體之任一者的一些實施例中，該第一抗體部分為Fab (「抗IL-13 Fab」)。在一些實施例中，該抗IL-13 Fab包含含有SEQ ID NO:124之胺基酸序列的第一多肽及含有SEQ ID NO:102或197之胺基酸序列的第二多肽。In some embodiments of any of the above multispecific constructs, the first antibody portion is a Fab ("anti-IL-13 Fab"). In some embodiments, the anti-IL-13 Fab comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197.

在根據上述多特異性構築體之任一者的一些實施例中，該第一抗體部分為全長抗體(「抗IL-13全長抗體」)。在一些實施例中，該抗IL-13全長抗體包含衍生自人類IgG (例如，人類IgG1、人類IgG2或人類IgG4)的Fc域。在一些實施例中，該Fc域係衍生自人類IgG1，其中：i)該Fc域之第一次單元及第二次單元各自含有SEQ ID NO:76-79之任一者的胺基酸序列；ii)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列；iii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列；iv)該Fc域之第一次單元含有SEQ ID NO:95之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:96之胺基酸序列；或v)該Fc域之第一次單元含有SEQ ID NO:96之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:95之胺基酸序列。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；ix)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或x)含有SEQ ID NO:130之胺基酸序列的第一重鏈、含有SEQ ID NO:131之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。In some embodiments according to any of the above multispecific constructs, the first antibody portion is a full-length antibody ("anti-IL-13 full-length antibody"). In some embodiments, the anti-IL-13 full-length antibody comprises an Fc domain derived from human IgG (e.g., human IgG1, human IgG2, or human IgG4). In some embodiments, the Fc domain is derived from human IgG1, wherein: i) the first and second subunits of the Fc domain each contain the amino acid sequence of any one of SEQ ID NOs: 76-79; ii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81; iii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80; iv) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 95, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 96; or v) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 96, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 95. In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 101 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 123 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 101 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; NO: 210 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207; vi) two heavy chains each containing the amino acid sequence of SEQ ID NO: 211 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206; vii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 212 and two light chains each containing the amino acid sequence of SEQ ID NO: 120 or 208; viii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 213 and two light chains each containing the amino acid sequence of SEQ ID NO: 120 or 208; ix) two heavy chains each containing the amino acid sequence of SEQ ID NO: 225 and two light chains each containing the amino acid sequence of SEQ ID NO: or x) a first heavy chain comprising the amino acid sequence of SEQ ID NO: 130, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 131, and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197.

在根據上述多特異性構築體之任一者的一些實施例中，該第二抗原為TSLP，其中該第二抗體部分包含重鏈可變區(VH2)及輕鏈可變區(VL2)，其中：(a)該VH2包含(i)含有SEQ ID NO:2之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:3之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3；(b)該VH2包含(i)含有SEQ ID NO:12之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:13之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3；(c)該VH2包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；(d)該VH2包含(i)含有SEQ ID NO:28之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:29之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:30之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:32之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:33之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:34之胺基酸序列的CDR-L3；(e)該VH2包含(i)含有SEQ ID NO:36之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:37之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:38之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:40之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:41之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:42之胺基酸序列的CDR-L3；或(f)該VH2包含(i)含有SEQ ID NO:44之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:45之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:46之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:48之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:49之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:50之胺基酸序列的CDR-L3。在一些實施例中，(a)該VH2含有SEQ ID NO:1之胺基酸序列，或與SEQ ID NO:1具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:5之胺基酸序列，或與SEQ ID NO:5具有至少約80%序列同一性的胺基酸序列；(b)該VH2含有SEQ ID NO:9之胺基酸序列，或與SEQ ID NO:9具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:10之胺基酸序列，或與SEQ ID NO:10具有至少約80%序列同一性的胺基酸序列；(c)該VH2含有SEQ ID NO:11之胺基酸序列，或與SEQ ID NO:11具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:15之胺基酸序列，或與SEQ ID NO:15具有至少約80%序列同一性的胺基酸序列；(d)該VH2含有SEQ ID NO:19之胺基酸序列，或與SEQ ID NO:19具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:23之胺基酸序列，或與SEQ ID NO:23具有至少約80%序列同一性的胺基酸序列；(e)該VH2含有SEQ ID NO:27之胺基酸序列，或與SEQ ID NO:27具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:31之胺基酸序列，或與SEQ ID NO:31具有至少約80%序列同一性的胺基酸序列；(f)該VH2含有SEQ ID NO:35之胺基酸序列，或與SEQ ID NO:35具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:39之胺基酸序列，或與SEQ ID NO:39具有至少約80%序列同一性的胺基酸序列；(g)該VH2含有SEQ ID NO:43之胺基酸序列，或與SEQ ID NO:43具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:47之胺基酸序列，或與SEQ ID NO:47具有至少約80%序列同一性的胺基酸序列；(h)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列；(i)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列；(j)該VH2含有SEQ ID NO:56之胺基酸序列，或與SEQ ID NO:56具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列；(k)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列；(l)該VH2含有SEQ ID NO:56之胺基酸序列，或與SEQ ID NO:56具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列；(m)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列；(n)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列；(o)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列；(p)該VH2含有SEQ ID NO:58之胺基酸序列，或與SEQ ID NO:58具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列；(q)該VH2含有SEQ ID NO:60之胺基酸序列，或與SEQ ID NO:60具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列；(r)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列；(s)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列；(t)該VH2含有SEQ ID NO:57之胺基酸序列，或與SEQ ID NO:57具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列；(u)該VH2含有SEQ ID NO:58之胺基酸序列，或與SEQ ID NO:58具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列；(v)該VH2含有SEQ ID NO:62之胺基酸序列，或與SEQ ID NO:62具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列；(w)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列；(x)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列；(y)該VH2含有SEQ ID NO:61之胺基酸序列，或與SEQ ID NO:61具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列；(z)該VH2含有SEQ ID NO:62之胺基酸序列，或與SEQ ID NO:62具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列；(aa)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列；(bb)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列；(cc)該VH2含有SEQ ID NO:188之胺基酸序列，或與SEQ ID NO:188具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:189之胺基酸序列，或與SEQ ID NO:189具有至少約80%序列同一性的胺基酸序列；(dd)該VH2含有SEQ ID NO:188之胺基酸序列，或與SEQ ID NO:188具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:190之胺基酸序列，或與SEQ ID NO:190具有至少約80%序列同一性的胺基酸序列；或(ee)該VH2含有SEQ ID NO:228之胺基酸序列，或與SEQ ID NO:228具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:190之胺基酸序列，或與SEQ ID NO:190具有至少約80%序列同一性的胺基酸序列。In some embodiments according to any of the above multispecific constructs, the second antigen is TSLP, wherein the second antibody portion comprises a heavy chain variable region (VH2) and a light chain variable region (VL2), wherein: (a) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 2; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 3; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 4; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 6; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 7; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 8; (b) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 12; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 3; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 4 NO:13; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:4; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:6; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:7; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:8; (c) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:22; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:25; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: NO:26; (d) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:28; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:29; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:30; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:32; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:33; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:34; (e) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:36; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:37; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:38. NO:38; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:40; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:41; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:42; or (f) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:44; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:45; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:46; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:48; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:49; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:50. In some embodiments, (a) the VH2 comprises the amino acid sequence of SEQ ID NO: 1, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 1, and the VL2 comprises the amino acid sequence of SEQ ID NO: 5, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 5; (b) the VH2 comprises the amino acid sequence of SEQ ID NO: 9, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 9, and the VL2 comprises the amino acid sequence of SEQ ID NO: 10, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 10; (c) the VH2 comprises the amino acid sequence of SEQ ID NO: 11, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 11, and the VL2 comprises the amino acid sequence of SEQ ID NO: 15, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: NO:15; (d) the VH2 comprises the amino acid sequence of SEQ ID NO:19, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:19, and the VL2 comprises the amino acid sequence of SEQ ID NO:23, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:23; (e) the VH2 comprises the amino acid sequence of SEQ ID NO:27, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:27, and the VL2 comprises the amino acid sequence of SEQ ID NO:31, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:31; (f) the VH2 comprises the amino acid sequence of SEQ ID NO:35, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:35, and the VL2 comprises the amino acid sequence of SEQ ID NO:39, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: NO:39 has an amino acid sequence with at least about 80% sequence identity; (g) the VH2 comprises the amino acid sequence of SEQ ID NO:43, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:43, and the VL2 comprises the amino acid sequence of SEQ ID NO:47, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:47; (h) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:53; (i) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: NO:54 has an amino acid sequence with at least about 80% sequence identity; (j) the VH2 contains the amino acid sequence of SEQ ID NO:56, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:56, and the VL2 contains the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:52; (k) the VH2 contains the amino acid sequence of SEQ ID NO:55, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:55, and the VL2 contains the amino acid sequence of SEQ ID NO:51, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:51; (l) the VH2 contains the amino acid sequence of SEQ ID NO:56, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:56, and the VL2 contains the amino acid sequence of SEQ ID NO:51, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: NO:51; (m) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:51, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:51; (n) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:51, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:51; (o) the VH2 comprises the amino acid sequence of SEQ ID NO:55, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:55, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: NO:52; (p) the VH2 comprises the amino acid sequence of SEQ ID NO:58, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:58, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:52; (q) the VH2 comprises the amino acid sequence of SEQ ID NO:60, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:60, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:52; (r) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: NO:52 has an amino acid sequence that is at least about 80% identical to SEQ ID NO:52; (s) the VH2 has an amino acid sequence of SEQ ID NO:55, or an amino acid sequence that is at least about 80% identical to SEQ ID NO:55, and the VL2 has an amino acid sequence of SEQ ID NO:53, or an amino acid sequence that is at least about 80% identical to SEQ ID NO:53; (t) the VH2 has an amino acid sequence of SEQ ID NO:57, or an amino acid sequence that is at least about 80% identical to SEQ ID NO:57, and the VL2 has an amino acid sequence of SEQ ID NO:53, or an amino acid sequence that is at least about 80% identical to SEQ ID NO:53; (u) the VH2 has an amino acid sequence of SEQ ID NO:58, or an amino acid sequence that is at least about 80% identical to SEQ ID NO:58, and the VL2 has an amino acid sequence of SEQ ID NO:53, or an amino acid sequence that is at least about 80% identical to SEQ ID NO: NO:53; (v) the VH2 comprises the amino acid sequence of SEQ ID NO:62, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:62, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:53; (w) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:53; (x) the VH2 comprises the amino acid sequence of SEQ ID NO:55, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:55, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: NO:54; (y) the VH2 comprises the amino acid sequence of SEQ ID NO:61, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:61, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:54; (z) the VH2 comprises the amino acid sequence of SEQ ID NO:62, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:62, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:54; (aa) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: (bb) the VH2 comprises the amino acid sequence of SEQ ID NO: 63, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: 63, and the VL2 comprises the amino acid sequence of SEQ ID NO: 52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: 52; (cc) the VH2 comprises the amino acid sequence of SEQ ID NO: 188, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: 188, and the VL2 comprises the amino acid sequence of SEQ ID NO: 189, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: 189; (dd) the VH2 comprises the amino acid sequence of SEQ ID NO: 188, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: 188, and the VL2 comprises the amino acid sequence of SEQ ID NO: NO: 190, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: 190; or (ee) the VH2 comprises the amino acid sequence of SEQ ID NO: 228, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: 228, and the VL2 comprises the amino acid sequence of SEQ ID NO: 190, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO: 190.

在根據上述多特異性構築體之任一者的一些實施例中，該第二抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、sdAb及scFv。In some embodiments according to any of the above multispecific constructs, the second antibody portion is selected from the group consisting of: a full-length antibody, Fab, Fab', F(ab')2 , sdAb, and scFv.

在根據上述多特異性構築體之任一者的一些實施例中，該第二抗體部分為全長抗體。在一些實施例中，該全長抗體包含衍生自人類IgG (例如，人類IgG1、人類IgG2或人類IgG4)的Fc域。在一些實施例中，該Fc域係衍生自人類IgG1，其中：i)該Fc域之第一次單元及第二次單元各自含有SEQ ID NO:76-79之任一者的胺基酸序列；ii)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列；或iii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列。在一些實施例中，該全長抗體為抗TSLP全長抗體，其包含：i)兩條各自含有SEQ ID NO:104之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈；或iii)含有SEQ ID NO:136之胺基酸序列的第一重鏈、含有SEQ ID NO:137之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈。In some embodiments according to any of the above multispecific constructs, the second antibody portion is a full-length antibody. In some embodiments, the full-length antibody comprises an Fc domain derived from human IgG (e.g., human IgG1, human IgG2, or human IgG4). In some embodiments, the Fc domain is derived from human IgG1, wherein: i) the first and second subunits of the Fc domain each contain the amino acid sequence of any one of SEQ ID NOs: 76-79; ii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81; or iii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80. In some embodiments, the full-length antibody is an anti-TSLP full-length antibody comprising: i) two heavy chains each containing the amino acid sequence of SEQ ID NO: 104 and two light chains each containing the amino acid sequence of SEQ ID NO: 103; ii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 105 and two light chains each containing the amino acid sequence of SEQ ID NO: 103; or iii) a first heavy chain containing the amino acid sequence of SEQ ID NO: 136, a second heavy chain containing the amino acid sequence of SEQ ID NO: 137, and two light chains each containing the amino acid sequence of SEQ ID NO: 103.

在根據上述多特異性構築體之任一者的一些實施例中，該第二抗體部分為scFv。在一些實施例中，該第二抗體部分為含有SEQ ID NO:106、107、218、219、226、227及229之任一者之胺基酸序列的抗TSLP scFv。In some embodiments according to any of the above multispecific constructs, the second antibody portion is a scFv. In some embodiments, the second antibody portion is an anti-TSLP scFv comprising the amino acid sequence of any one of SEQ ID NOs: 106, 107, 218, 219, 226, 227, and 229.

在根據上述多特異性構築體之任一者的一些實施例中，該第二抗體部分為Fab。在一些實施例中，該第二抗體部分為抗TSLP Fab，其中該抗TSLP Fab包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。In some embodiments according to any of the above multispecific constructs, the second antibody portion is a Fab. In some embodiments, the second antibody portion is an anti-TSLP Fab, wherein the anti-TSLP Fab comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103.

在根據上述多特異性構築體之任一者的一些實施例中，該第一抗體部分包含重鏈(H1)及輕鏈(L1)，其中該H1包含VH1及H1-CH1，其中該L1包含VL1及L1-CL，且其中：(i)該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)；及/或(ii)該H1包含位於位置126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該H1包含位於F170、S183及V185處的取代(EU編號)，且該L1包含位於L135處的取代(EU編號)。在一些實施例中，該F170取代為F170I或F170V、該S183取代為S183L或S183I且該V185取代為V185L。在一些實施例中，該L135取代為L135F。在一些實施例中，(a)該H1包含F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)；或(b)該H1包含F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該λ輕鏈含有SEQ ID NO:74或75之胺基酸序列。在一些實施例中，該H1包含位於F126及C220處的取代(EU編號)，且該L1包含位於E124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該κ輕鏈含有SEQ ID NO:73之胺基酸序列。在一些實施例中，該H1包含位於F126及C220處的取代(EU編號)，且該L1包含位於Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，(i)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代；(ii)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代；(iii)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含Q124C、L135F及C214S取代；或(iv)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含Q124C、L135F及C214S取代；其中該位置係根據EU編號。In some embodiments according to any of the above multispecific constructs, the first antibody portion comprises a heavy chain (H1) and a light chain (L1), wherein the H1 comprises VH1 and H1-CH1, wherein the L1 comprises VL1 and L1-CL, and wherein: (i) the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering); and/or (ii) the H1 comprises substitutions at positions 126 and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the H1 comprises substitutions at F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at L135 (EU numbering). In some embodiments, F170 is substituted with F170I or F170V, S183 is substituted with S183L or S183I, and V185 is substituted with V185L. In some embodiments, L135 is substituted with L135F. In some embodiments, (a) H1 comprises F170I, S183L, and V185L substitutions (EU numbering), and L1 comprises L135F substitution (EU numbering); or (b) H1 comprises F170V, S183I, and V185L substitutions (EU numbering), and L1 comprises L135F substitution (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, the lambda light chain comprises the amino acid sequence of SEQ ID NO: 74 or 75. In some embodiments, the H1 comprises substitutions at F126 and C220 (EU numbering), and the L1 comprises substitutions at E124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at F126C and C220S (EU numbering), and the L1 comprises substitutions at E124C and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the kappa light chain comprises the amino acid sequence of SEQ ID NO: 73. In some embodiments, the H1 comprises substitutions at F126 and C220 (EU numbering), and the L1 comprises substitutions at Q124 and C214 (EU numbering). In some embodiments, the H1 comprises F126C and C220S substitutions (EU numbering), and the L1 comprises Q124C and C214S substitutions (EU numbering). In some embodiments, (i) the H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and the L1 comprises E124C, L135F, and C214S substitutions; (ii) the H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and the L1 comprises E124C, L135F, and C214S substitutions; (iii) ) the H1 comprises F126C, F170I, S183L, V185L and C220S substitutions, and the L1 comprises Q124C, L135F and C214S substitutions; or (iv) the H1 comprises F126C, F170V, S183I, V185L and C220S substitutions, and the L1 comprises Q124C, L135F and C214S substitutions; wherein the positions are according to EU numbering.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體進一步包含Fc域，其中該Fc域包含第一次單元及第二次單元。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4。在一些實施例中，該Fc域係衍生自IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元含有SEQ ID NO:77之胺基酸序列。在一些實施例中，該Fc域包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元含有SEQ ID NO:78之胺基酸序列。在一些實施例中，該Fc域包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，該Fc域之每一次單元含有SEQ ID NO:79之胺基酸序列。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列；(ii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列；(iii)該Fc域之第一次單元含有SEQ ID NO:95之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:96之胺基酸序列；或(iv)該Fc域之第一次單元含有SEQ ID NO:96之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:95之胺基酸序列。In some embodiments according to any of the above multispecific constructs, the multispecific construct further comprises an Fc domain, wherein the Fc domain comprises a first subunit and a second subunit. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4. In some embodiments, the Fc domain is derived from IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO: 77. In some embodiments, the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO: 78. In some embodiments, the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO: 79. In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (i) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81; (ii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80; (iii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 95, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 96; or (iv) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 96, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 95.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1、H1-CH1及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2、H2-CH1及該Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；且其中H1及L1形成抗IL-13抗體部分，且H2及L2形成該第二抗體部分。在一些實施例中，該Fc域係衍生自人類IgG1。在一些實施例中，該L1-CL係衍生自人類λ輕鏈。在一些實施例中，(a)該H1包含F170I、S183L及V185L取代，且該L1包含L135F取代；(b)該H1包含F170V、S183I及V185L取代，且該L1包含L135F取代；(c)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代；(d)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代；或(e)該H1包含F126C及C220S取代，且該L1包含E124C及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，(a)該H1包含F170I、S183L、V185L、T366S、L368A及Y407V取代，該L1包含L135F取代，且該H2包含T366W取代；(b)該H1包含F170I、S183L、V185L及T366W取代，該L1包含L135F取代，且該H2包含T366S、L368A及Y407V取代；(c)該H1包含F170V、S183I、V185L、T366S、L368A及Y407V取代，該L1包含L135F取代，且該H2包含T366W取代；(d)該H1包含F170V、S183I、V185L及T366W取代，該L1包含L135F取代，且該H2包含T366S、L368A及Y407V取代；(e)該H1包含F126C、F170I、S183L、V185L、C220S、T366S、L368A及Y407V取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366W取代；(f)該H1包含F126C、F170I、S183L、V185L、C220S及T366W取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366S、L368A及Y407V取代；(g)該H1包含F126C、F170V、S183I、V185L、C220S、T366S、L368A及Y407V取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366W取代；(h)該H1包含F126C、F170V、S183I、V185L、C220S及T366W取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366S、L368A及Y407V取代；(i)該H1包含F126C、C220S、T366S、L368A及Y407V取代，該L1包含E124C及C214S取代，且該H2包含T366W取代；或(j)該H1包含F126C、C220S及T366W取代，該L1包含E124C及C214S取代，且該H2包含T366S、L368A及Y407V取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，(a)該H1包含F170I、S183L、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代；(b)該H1包含F170I、S183L、V185L、L234A、L235A、T366W、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；(c)該H1包含F170V、S183I、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代；(d)該H1包含F170V、S183I、V185L、L234A、L235A、T366W、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；(e)該H1包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代；(f)該H1包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；(g)該H1包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代；(h)該H1包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；(i)該H1包含F126C、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代；或(j)該H1包含F126C、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，(a)該H1含有SEQ ID NO:87之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列；(b)該H1含有SEQ ID NO:88之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列；(c)該H1含有SEQ ID NO:89之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列；(d)該H1含有SEQ ID NO:90之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列；(e)該H1含有SEQ ID NO:91之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列；(f)該H1含有SEQ ID NO:92之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列；(g)該H1含有SEQ ID NO:93之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列；(h)該H1含有SEQ ID NO:94之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列；(i)該H1含有SEQ ID NO:198之胺基酸序列，該L1含有SEQ ID NO:204或205之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列；或(j)該H1含有SEQ ID NO:199之胺基酸序列，該L1含有SEQ ID NO:204或205之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列。在一些實施例中，該L2-CL係衍生自人類κ輕鏈。在一些實施例中，該第二抗原為TSLP。在一些實施例中，(a)該H1含有SEQ ID NO:139之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；(b)該H1含有SEQ ID NO:138之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；(c)該H1含有SEQ ID NO:141之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；(d)該H1含有SEQ ID NO:140之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；(e)該H1含有SEQ ID NO:126之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；(f)該H1含有SEQ ID NO:127之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；(g)該H1含有SEQ ID NO:128之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；(h)該H1含有SEQ ID NO:129之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；(i)該H1含有SEQ ID NO:200之胺基酸序列，該L1含有SEQ ID NO:121或206之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；或(j)該H1含有SEQ ID NO:201之胺基酸序列，該L1含有SEQ ID NO:121或206之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。在一些實施例中，該Fc域係衍生自人類IgG1。在一些實施例中，該L2-CL係衍生自人類λ輕鏈。在一些實施例中，(a)該H2包含F170I、S183L及V185L取代，且該L2包含L135F取代；(b)該H2包含F170V、S183I及V185L取代，且該L2包含L135F取代；(c)該H2包含F126C、F170I、S183L、V185L及C220S取代，且該L2包含E124C、L135F及C214S取代；(d)該H2包含F126C、F170V、S183I、V185L及C220S取代，且該L2包含E124C、L135F及C214S取代；或(e)該H2包含F126C及C220S取代，且該L2包含E124C及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，(a)該H2包含F170I、S183L、V185L、T366S、L368A及Y407V取代，該L2包含L135F取代，且該H1包含T366W取代；(b)該H2包含F170I、S183L、V185L及T366W取代，該L2包含L135F取代，且該H1包含T366S、L368A及Y407V取代；(c)該H2包含F170V、S183I、V185L、T366S、L368A及Y407V取代，該L2包含L135F取代，且該H1包含T366W取代；(d)該H2包含F170V、S183I、V185L及T366W取代，該L2包含L135F取代，且該H1包含T366S、L368A及Y407V取代；(e)該H2包含F126C、F170I、S183L、V185L、C220S、T366S、L368A及Y407V取代，該L2包含E124C、L135F及C214S取代，且該H1包含T366W取代；(f)該H2包含F126C、F170I、S183L、V185L、C220S及T366W取代，該L2包含E124C、L135F及C214S取代，且該H1包含T366S、L368A及Y407V取代；(g)該H2包含F126C、F170V、S183I、V185L、C220S、T366S、L368A及Y407V取代，該L2包含E124C、L135F及C214S取代，且該H1包含T366W取代；(h)該H2包含F126C、F170V、S183I、V185L、C220S及T366W取代，該L2包含E124C、L135F及C214S取代，且該H1包含T366S、L368A及Y407V取代；(i)該H2包含F126C、C220S、T366S、L368A及Y407V取代，該L2包含E124C及C214S取代，且該H1包含T366W取代；或(j)該H2包含F126C、C220S及T366W取代，該L2包含E124C及C214S取代，且該H1包含T366S、L368A及Y407V取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，(a)該H2包含F170I、S183L、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含L135F取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代；(b)該H2包含F170I、S183L、V185L、L234A、L235A、T366W、M428L及N434S取代，該L2包含L135F取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；(c)該H2包含F170V、S183I、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含L135F取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代；(d)該H2包含F170V、S183I、V185L、L234A、L235A、T366W、M428L及N434S取代，該L2包含L135F取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；(e)該H2包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含E124C、L135F及C214S取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代；(f)該H2包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L2包含E124C、L135F及C214S取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；(g)該H2包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含E124C、L135F及C214S取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代；(h)該H2包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L2包含E124C、L135F及C214S取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；(i)該H2包含F126C、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含E124C及C214S取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代；或(j)該H2包含F126C、C220S、L234A、L235A、T366W、M428L及N434S取代，該L2包含E124C及C214S取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，(a)該H2含有SEQ ID NO:87之胺基酸序列，該L2含有SEQ ID NO:173或202之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列；(b)該H2含有SEQ ID NO:88之胺基酸序列，該L2含有SEQ ID NO:173或202之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列；(c)該H2含有SEQ ID NO:89之胺基酸序列，該L2含有SEQ ID NO:173或202之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列；(d)該H2含有SEQ ID NO:90之胺基酸序列，該L2含有SEQ ID NO:173或202之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列；(e)該H2含有SEQ ID NO:91之胺基酸序列，該L2含有SEQ ID NO:174或203之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列；(f)該H2含有SEQ ID NO:92之胺基酸序列，該L2含有SEQ ID NO:174或203之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列；(g)該H2含有SEQ ID NO:93之胺基酸序列，該L2含有SEQ ID NO:174或203之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列；(h)該H2含有SEQ ID NO:94之胺基酸序列，該L2含有SEQ ID NO:174或203之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列；(i)該H2含有SEQ ID NO:198之胺基酸序列，該L2含有SEQ ID NO:204或205之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列；或(j)該H2含有SEQ ID NO:199之胺基酸序列，該L2含有SEQ ID NO:204或205之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列。在一些實施例中，該L1-CL係衍生自人類κ輕鏈。在一些實施例中，該第二抗原為TSLP。In some embodiments of any of the above multispecific constructs, the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and the Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form the anti-IL-13 antibody portion, and H2 and L2 form the second antibody portion. In some embodiments, the Fc domain is derived from human IgG1. In some embodiments, the L1-CL is derived from a human lambda light chain. In some embodiments, (a) the H1 comprises F170I, S183L, and V185L substitutions, and the L1 comprises L135F substitutions; (b) the H1 comprises F170V, S183I, and V185L substitutions, and the L1 comprises L135F substitutions; (c) the H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and the L1 comprises E124 C, L135F and C214S substitutions; (d) the H1 comprises F126C, F170V, S183I, V185L and C220S substitutions, and the L1 comprises E124C, L135F and C214S substitutions; or (e) the H1 comprises F126C and C220S substitutions, and the L1 comprises E124C and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, (a) the H1 comprises F170I, S183L, V185L, T366S, L368A, and Y407V substitutions, the L1 comprises an L135F substitution, and the H2 comprises a T366W substitution; (b) the H1 comprises F170I, S183L, V185L, and T366W substitutions, the L1 comprises an L135F substitution, and the H2 comprises T366S, L368A, and Y407V substitutions; (c) the H1 comprises F170V, S183I, V185L, T366S, L368A, and Y407V substitutions, the L1 comprises an L135F substitution, and the H2 comprises a T366W substitution; (d) the H1 comprises F170V, S183I, V185L and T366W substitutions, the L1 comprises L135F substitutions, and the H2 comprises T366S, L368A and Y407V substitutions; (e) the H1 comprises F126C, F170I, S183L, V185L, C220S, T366S, L368A and Y407V substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises a T366W substitution; (f) the H1 comprises F126C, F170I, S183L, V185L and T366W substitutions, the L1 comprises L135F substitutions, and the H2 comprises T366S, L368A and Y407V substitutions (g) the H1 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A and Y407V substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises T366W substitutions; (h) the H1 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A and Y407V substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises T366W substitutions 1 comprises E124C, L135F and C214S substitutions, and the H2 comprises T366S, L368A and Y407V substitutions; (i) the H1 comprises F126C, C220S, T366S, L368A and Y407V substitutions, the L1 comprises E124C and C214S substitutions, and the H2 comprises T366W substitution; or (j) the H1 comprises F126C, C220S and T366W substitutions, the L1 comprises E124C and C214S substitutions, and the H2 comprises T366S, L368A and Y407V substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, (a) the H1 comprises F170I, S183L, V185L, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, the L1 comprises L135F substitutions, and the H2 comprises L234A, L235A, T366W, M428L, and N434S substitutions; (b) the H1 comprises F170I, S183L, V185L, L234A, L235A, , T366W, M428L and N434S substitutions, said L1 comprises an L135F substitution, and said H2 comprises an L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (c) said H1 comprises an F170V, S183I, V185L, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, said L1 comprises an L135F substitution, and the H2 comprises L234A, L235A, T366W, M428L and N434S substitutions; (d) the H1 comprises F170V, S183I, V185L, L234A, L235A, T366W, M428L and N434S substitutions, the L1 comprises L135F substitution, and the H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (e) the H1 comprises containing F126C, F170I, S183L, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, said L1 containing E124C, L135F and C214S substitutions, and said H2 containing L234A, L235A, T366W, M428L and N434S substitutions; (f) said H1 containing F126C, F170I, S183L, V185L 5L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, said L1 comprises E124C, L135F and C214S substitutions, and said H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (g) said H1 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises L234A, L235A, T366W, M428L and N434S substitutions; (h) the H1 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, the L 1 comprises E124C, L135F and C214S substitutions, and said H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (i) said H1 comprises F126C, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, said L1 comprises E124C and C214S substitutions, and said H2 comprises L234 A, L235A, T366W, M428L and N434S substitutions; or (j) the H1 comprises F126C, C220S, L234A, L235A, T366W, M428L and N434S substitutions, the L1 comprises E124C and C214S substitutions, and the H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, (a) the H1 contains the amino acid sequence of SEQ ID NO: 87, the L1 contains the amino acid sequence of SEQ ID NO: 173 or 202, and the H2 contains the amino acid sequence of SEQ ID NO: 85; (b) the H1 contains the amino acid sequence of SEQ ID NO: 88, the L1 contains the amino acid sequence of SEQ ID NO: 173 or 202, and the H2 contains the amino acid sequence of SEQ ID NO: 86; (c) the H1 contains the amino acid sequence of SEQ ID NO: 89, the L1 contains the amino acid sequence of SEQ ID NO: 173 or 202, and the H2 contains the amino acid sequence of SEQ ID NO: 85; (d) the H1 contains the amino acid sequence of SEQ ID NO: 90, the L1 contains the amino acid sequence of SEQ ID NO: 173 or 202, and the H2 contains the amino acid sequence of SEQ ID NO: 86; (e) the H1 contains the amino acid sequence of SEQ ID NO: NO:91, the L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H2 contains the amino acid sequence of SEQ ID NO:85; (f) the H1 contains the amino acid sequence of SEQ ID NO:92, the L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H2 contains the amino acid sequence of SEQ ID NO:86; (g) the H1 contains the amino acid sequence of SEQ ID NO:93, the L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H2 contains the amino acid sequence of SEQ ID NO:85; (h) the H1 contains the amino acid sequence of SEQ ID NO:94, the L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H2 contains the amino acid sequence of SEQ ID NO:86; (i) the H1 contains the amino acid sequence of SEQ ID NO:198, the L1 contains the amino acid sequence of SEQ ID NO: (j) the H1 contains the amino acid sequence of SEQ ID NO: 199, the L1 contains the amino acid sequence of SEQ ID NO: 204 or 205, and the H2 contains the amino acid sequence of SEQ ID NO: 86. In some embodiments, the L2-CL is derived from a human kappa light chain. In some embodiments, the second antigen is TSLP. In some embodiments, (a) the H1 contains the amino acid sequence of SEQ ID NO: 139, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (b) the H1 contains the amino acid sequence of SEQ ID NO: 138, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (c) the H1 contains the amino acid sequence of SEQ ID NO: 141, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (d) the H1 contains the amino acid sequence of SEQ ID NO: (e) the H1 contains the amino acid sequence of SEQ ID NO: 126, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (f) the H1 contains the amino acid sequence of SEQ ID NO: 127, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (g) the H1 contains the amino acid sequence of SEQ ID NO: 128, the L1 contains the amino acid sequence of SEQ ID NO: 129, NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (h) the H1 contains the amino acid sequence of SEQ ID NO: 129, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (i) the H1 contains the amino acid sequence of SEQ ID NO: 200, the L1 contains the amino acid sequence of SEQ ID NO: 121 or 206, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; or (j) the H1 contains the amino acid sequence of SEQ ID NO: 201, the L1 contains the amino acid sequence of SEQ ID NO: NO:121 or 206, the H2 contains the amino acid sequence of SEQ ID NO:137, and the L2 contains the amino acid sequence of SEQ ID NO:103. In some embodiments, the Fc domain is derived from human IgG1. In some embodiments, the L2-CL is derived from a human lambda light chain. In some embodiments, (a) the H2 contains F170I, S183L, and V185L substitutions, and the L2 contains an L135F substitution; (b) the H2 contains F170V, S183I, and V185L substitutions, and the L2 contains an L135F substitution; (c) the H2 contains F126C, F170I, S183L, V185L, and C220S substitutions, and the L2 contains E124 C, L135F and C214S substitutions; (d) the H2 comprises F126C, F170V, S183I, V185L and C220S substitutions, and the L2 comprises E124C, L135F and C214S substitutions; or (e) the H2 comprises F126C and C220S substitutions, and the L2 comprises E124C and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, (a) the H2 comprises F170I, S183L, V185L, T366S, L368A, and Y407V substitutions, the L2 comprises an L135F substitution, and the H1 comprises a T366W substitution; (b) the H2 comprises F170I, S183L, V185L, and T366W substitutions, the L2 comprises an L135F substitution, and the H1 comprises T366S, L368A, and Y407V substitutions; (c) the H2 comprises F170V, S183I, V185L, T366S, L368A, and Y407V substitutions, the L2 comprises an L135F substitution, and the H1 comprises a T366W substitution; (d) the H2 comprises a F170V, S183I, V185L and T366W substitution, the L2 comprises a L135F substitution, and the H1 comprises a T366S, L368A and Y407V substitution; (e) the H2 comprises a F126C, F170I, S183L, V185L, C220S, T366S, L368A and Y407V substitution, the L2 comprises an E124C, L135F and C214S substitution, and the H1 comprises a T366W substitution; (f) the H2 comprises a F126C, F170I, S183L, V185L, (g) the H2 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A and Y407V substitutions, the L2 comprises E124C, L135F and C214S substitutions, and the H1 comprises T366W substitutions; (h) the H2 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A and Y407V substitutions, the L2 comprises E124C, L135F and C214S substitutions, and the H1 comprises T366W substitutions. 2 comprises E124C, L135F and C214S substitutions, and the H1 comprises T366S, L368A and Y407V substitutions; (i) the H2 comprises F126C, C220S, T366S, L368A and Y407V substitutions, the L2 comprises E124C and C214S substitutions, and the H1 comprises T366W substitution; or (j) the H2 comprises F126C, C220S and T366W substitutions, the L2 comprises E124C and C214S substitutions, and the H1 comprises T366S, L368A and Y407V substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, (a) the H2 comprises F170I, S183L, V185L, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, the L2 comprises L135F substitutions, and the H1 comprises L234A, L235A, T366W, M428L, and N434S substitutions; (b) the H2 comprises F170I, S183L, V185L, L234A, L235A, , T366W, M428L and N434S substitutions, said L2 comprises an L135F substitution, and said H1 comprises an L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (c) said H2 comprises an F170V, S183I, V185L, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, said L2 comprises an L135F substitution, and the H1 comprises L234A, L235A, T366W, M428L and N434S substitutions; (d) the H2 comprises F170V, S183I, V185L, L234A, L235A, T366W, M428L and N434S substitutions, the L2 comprises L135F substitution, and the H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (e) the H2 comprises (f) wherein H2 comprises F126C, F170I, S183L, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, wherein L2 comprises E124C, L135F and C214S substitutions, and wherein H1 comprises L234A, L235A, T366W, M428L and N434S substitutions; (g) said H2 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, said L2 comprises E124C, L135F and C214S substitutions, and said H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; T366S, L368A, Y407V, M428L and N434S substitutions, the L2 comprises E124C, L135F and C214S substitutions, and the H1 comprises L234A, L235A, T366W, M428L and N434S substitutions; (h) the H2 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, the L 2 comprises E124C, L135F and C214S substitutions, and the H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (i) the H2 comprises F126C, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, the L2 comprises E124C and C214S substitutions, and the H1 comprises L234 A, L235A, T366W, M428L and N434S substitutions; or (j) the H2 comprises F126C, C220S, L234A, L235A, T366W, M428L and N434S substitutions, the L2 comprises E124C and C214S substitutions, and the H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, (a) the H2 contains the amino acid sequence of SEQ ID NO: 87, the L2 contains the amino acid sequence of SEQ ID NO: 173 or 202, and the H1 contains the amino acid sequence of SEQ ID NO: 85; (b) the H2 contains the amino acid sequence of SEQ ID NO: 88, the L2 contains the amino acid sequence of SEQ ID NO: 173 or 202, and the H1 contains the amino acid sequence of SEQ ID NO: 86; (c) the H2 contains the amino acid sequence of SEQ ID NO: 89, the L2 contains the amino acid sequence of SEQ ID NO: 173 or 202, and the H1 contains the amino acid sequence of SEQ ID NO: 85; (d) the H2 contains the amino acid sequence of SEQ ID NO: 90, the L2 contains the amino acid sequence of SEQ ID NO: 173 or 202, and the H1 contains the amino acid sequence of SEQ ID NO: 86; (e) the H2 contains the amino acid sequence of SEQ ID NO: NO:91, the L2 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H1 contains the amino acid sequence of SEQ ID NO:85; (f) the H2 contains the amino acid sequence of SEQ ID NO:92, the L2 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H1 contains the amino acid sequence of SEQ ID NO:86; (g) the H2 contains the amino acid sequence of SEQ ID NO:93, the L2 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H1 contains the amino acid sequence of SEQ ID NO:85; (h) the H2 contains the amino acid sequence of SEQ ID NO:94, the L2 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H1 contains the amino acid sequence of SEQ ID NO:86; (i) the H2 contains the amino acid sequence of SEQ ID NO:198, the L2 contains the amino acid sequence of SEQ ID NO: (j) the H2 comprises the amino acid sequence of SEQ ID NO: 199, the L2 comprises the amino acid sequence of SEQ ID NO: 204 or 205, and the H1 comprises the amino acid sequence of SEQ ID NO: 86. In some embodiments, the L1-CL is derived from a human kappa light chain. In some embodiments, the second antigen is TSLP.

在根據上述多特異性構築體之任一者的一些實施例中，該特異性地結合至IL-13的第一抗體部分與該特異性地結合至第二標靶抗原的第二抗體部分係經由連接子彼此融合。在一些實施例中，該連接子含有GG及SEQ ID NO:98-99之任一者的胺基酸序列。In some embodiments of any of the above multispecific constructs, the first antibody portion that specifically binds to IL-13 and the second antibody portion that specifically binds to a second target antigen are fused to each other via a linker. In some embodiments, the linker comprises GG and the amino acid sequence of any one of SEQ ID NOs: 98-99.

在根據上述多特異性構築體之任一者的一些實施例中，該第一抗體部分為scFv (「抗IL-13 scFv」)，其中該第二抗體部分特異性地結合至第二標靶抗原的全長抗體，且其中該抗IL-13 scFv係融合至第二抗體部分之重鏈之一者的C端以形成融合多肽。In some embodiments according to any of the above multispecific constructs, the first antibody portion is a scFv ("anti-IL-13 scFv"), wherein the second antibody portion is a full-length antibody that specifically binds to a second target antigen, and wherein the anti-IL-13 scFv is fused to the C-terminus of one of the heavy chains of the second antibody portion to form a fusion polypeptide.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體包含兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)，且該第二抗體部分為特異性地結合第二標靶抗原的全長抗體；且其中抗IL-13 scFv1係融合至第二抗體部分之第一重鏈的C端以形成第一融合多肽，且抗IL-13 scFv2係融合至第二抗體部分之第二重鏈的C端以形成第二融合多肽。在一些實施例中，該第二標靶抗原為TSLP (亦即，該第二抗體部分為「抗TSLP全長抗體」)。在一些實施例中，該抗TSLP全長抗體包含兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈及兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:113之胺基酸序列。In some embodiments of any of the above multispecific constructs, the multispecific construct comprises two anti-IL-13 antibody portions, each of which is a scFv ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), and the second antibody portion is a full-length antibody that specifically binds to a second target antigen; wherein the anti-IL-13 scFv1 is fused to the C-terminus of the first heavy chain of the second antibody portion to form a first fusion polypeptide, and the anti-IL-13 scFv2 is fused to the C-terminus of the second heavy chain of the second antibody portion to form a second fusion polypeptide. In some embodiments, the second target antigen is TSLP (i.e., the second antibody portion is an "anti-TSLP full-length antibody"). In some embodiments, the anti-TSLP full-length antibody comprises two light chains, each containing the amino acid sequence of SEQ ID NO: 103, and two heavy chains, each containing the amino acid sequence of SEQ ID NO: 105, and wherein the first fusion polypeptide and the second fusion polypeptide each contain the amino acid sequence of SEQ ID NO: 113.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體包含兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)、兩個第二抗體部分，其等為特異性地結合該第二標靶抗原的Fab (「Fab1」及「Fab2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-抗IL-13 scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-抗IL-13 scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成Fab2。在一些實施例中，該第二標靶抗原為TSLP。在一些實施例中，該第一多肽及該第四多肽各自含有SEQ ID NO:103之胺基酸序列，且該第二多肽及該第三多肽各自含有SEQ ID NO:110之胺基酸序列。In some embodiments according to any of the above multispecific constructs, the multispecific construct comprises two anti-IL-13 antibody portions, which are scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), two second antibody portions, which are Fabs that specifically bind to the second target antigen ("Fab1" and "Fab2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a first polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-anti-IL-13 The invention further comprises a second polypeptide comprising: scFv1-optional linker-Fc domain; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optional linker-anti-IL-13 scFv2-optional linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form Fab2. In some embodiments, the second target antigen is TSLP. In some embodiments, the first polypeptide and the fourth polypeptide each comprise the amino acid sequence of SEQ ID NO: 103, and the second polypeptide and the third polypeptide each comprise the amino acid sequence of SEQ ID NO: 110.

在根據上述多特異性構築體之任一者的一些實施例中，該第一抗體部分為全長抗體(「抗IL-13全長抗體」)，其中該第二抗體部分為特異性地結合至該第二標靶抗原的scFv，且其中該scFv係融合至該抗IL-13全長抗體之重鏈之一者的C端以形成融合多肽。In some embodiments according to any of the above multispecific constructs, the first antibody portion is a full-length antibody ("anti-IL-13 full-length antibody"), wherein the second antibody portion is a scFv that specifically binds to the second target antigen, and wherein the scFv is fused to the C-terminus of one of the heavy chains of the anti-IL-13 full-length antibody to form a fusion polypeptide.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體包含該第一抗體部分，其為抗IL-13全長抗體，以及兩個第二抗體部分，其等為特異性地結合該第二標靶抗原的scFv (「scFv1」及「scFv2」)；且其中scFv1係融合至該抗IL-13全長抗體之第一重鏈的C端以形成第一融合多肽，且scFv2係融合至該抗IL-13全長抗體之第二重鏈的C端以形成第二融合多肽。在一些實施例中，該第二標靶抗原為TSLP。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:111之胺基酸序列。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:220-223之任一者的胺基酸序列。In some embodiments of any of the above multispecific constructs, the multispecific construct comprises the first antibody portion, which is a full-length anti-IL-13 antibody, and two second antibody portions, which are scFvs ("scFv1" and "scFv2") that specifically bind to the second target antigen; wherein scFv1 is fused to the C-terminus of the first heavy chain of the full-length anti-IL-13 antibody to form a first fusion polypeptide, and scFv2 is fused to the C-terminus of the second heavy chain of the full-length anti-IL-13 antibody to form a second fusion polypeptide. In some embodiments, the second target antigen is TSLP. In some embodiments, the full-length anti-IL-13 antibody comprises two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197, and wherein the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of SEQ ID NO: 111. In some embodiments, the full-length anti-IL-13 antibody comprises two heavy chains each comprising the amino acid sequence of SEQ ID NO: 225 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197, and wherein the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of any one of SEQ ID NOs: 220-223.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體包含兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)、兩個第二抗體部分，其等為特異性地結合第二標靶抗原的scFv (「scFv1」及「scFv2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；且其中VH1-(H1-CH1)及VL1-(L1-CL)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2。在一些實施例中，該第二標靶抗原為TSLP。在一些實施例中，該第一多肽及該第四多肽各自含有SEQ ID NO:102或197之胺基酸序列，且該第二多肽及該第三多肽各自含有SEQ ID NO:112之胺基酸序列。In some embodiments according to any of the above multispecific constructs, the multispecific construct comprises two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2"), two second antibody portions, which are scFvs that specifically bind to a second target antigen. ("scFv1" and "scFv2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optional linker-scFv1-optional linker-Fc domain; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optional linker-scFv2-optional linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); and wherein VH1-(H1-CH1) and VL1-(L1-CL) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-IL-13 Fab2. In some embodiments, the second target antigen is TSLP. In some embodiments, the first polypeptide and the fourth polypeptide each contain the amino acid sequence of SEQ ID NO: 102 or 197, and the second polypeptide and the third polypeptide each contain the amino acid sequence of SEQ ID NO: 112.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性包含兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)，以及第二抗體部分，其為特異性地結合至第二標靶抗原的全長抗體；其中該多特異性構築體包含：i)含有第二抗體部分之第一輕鏈的第一多肽；ii)含有N’至C’：第二抗體部分之第一重鏈-視情況的連接子-VH1-(H1-CH1)的第二多肽；iii)含有N’至C’：第二抗體部分之第二重鏈-視情況的連接子-VH2-(H2-CH1)的第三多肽；iv)含有第二抗體部分之第二輕鏈的第四多肽；v)含有N’至C’：VL1-(L1-CL)的第五多肽；及vi)含有N’至C’：VL2-(L2-CL)的第六多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VL2-(L2-CL)及VH2-(H2-CH1)形成抗IL-13 Fab2。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；或(e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該第二標靶抗原為TSLP (亦即，該第二抗體部分為「抗TSLP全長抗體」)。在一些實施例中，該抗TSLP全長抗體包含兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈。In some embodiments according to any of the above multispecific constructs, the multispecific construct comprises two anti-IL-13 antibody portions, which are Fab ("anti-IL-13 Fab1" and "anti-IL-13 Fab2")), and a second antibody portion that is a full-length antibody that specifically binds to a second target antigen; wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain of the second antibody portion; ii) a second polypeptide comprising N' to C': the first heavy chain of the second antibody portion - optionally a linker - VH1-(H1-CH1); iii) a third polypeptide comprising N' to C': the second heavy chain of the second antibody portion - optionally a linker - VH2-(H2-CH1); iv) a fourth polypeptide comprising the second light chain of the second antibody portion; v) a fifth polypeptide comprising N' to C': VL1-(L1-CL); and vi) a sixth polypeptide comprising N' to C': VL2-(L2-CL); and wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 Fab1, and VL2-(L2-CL) and VH2-(H2-CH1) form anti-IL-13 Fab2. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) (d) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; or (e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the second target antigen is TSLP (i.e., the second antibody portion is an "anti-TSLP full-length antibody"). In some embodiments, the anti-TSLP full-length antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 105, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 103.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體包含抗IL-13抗體部分，其為全長抗體(「抗IL-13全長抗體」)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原的Fab (「Fab1」及「Fab2」)；其中該多特異性構築體包含：i)含有抗IL-13全長抗體之第一輕鏈(L1)的第一多肽；ii)含有N’至C’：抗IL-13全長抗體之第一重鏈(H1)-視情況的連接子-VH3-(H3-CH1)的第二多肽；iii)含有N’至C’：抗IL-13全長抗體之第二重鏈(H2)-視情況的連接子-VH4-(H4-CH1)的第三多肽；iv)含有抗IL-13全長抗體之第二輕鏈(L2)的第四多肽；v)含有N’至C’：VL3-(L3-CL)的第五多肽；及vi)含有N’至C’：VL4-(L4-CL)的第六多肽；且其中VH3-(H3-CH1)及VL3-(L3-CL)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；或(e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或ix)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈。在一些實施例中，該第二標靶抗原為TSLP (亦即，該兩個抗體部分為「抗TSLP Fab1」及「抗TSLP Fab2」)。在一些實施例中，該抗TSLP Fab1及該抗TSLP Fab2各自包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。In some embodiments according to any of the above multispecific constructs, the multispecific construct comprises an anti-IL-13 antibody portion that is a full-length antibody ("anti-IL-13 full-length antibody"), and two second antibody portions that are Fabs that specifically bind to a second target antigen ("Fab1" and "Fab2"); wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain (L1) of the anti-IL-13 full-length antibody; ii) a second polypeptide comprising N' to C': the first heavy chain (H1) of the anti-IL-13 full-length antibody -optionally a linker-VH3-(H3-CH1); iii) a second polypeptide comprising N' to C': the second heavy chain (H2) of the anti-IL-13 full-length antibody -optionally a linker-VH4-(H4-CH1); -CH1); iv) a fourth polypeptide containing the second light chain (L2) of the anti-IL-13 full-length antibody; v) a fifth polypeptide containing N' to C': VL3-(L3-CL); and vi) a sixth polypeptide containing N' to C': VL4-(L4-CL); and wherein VH3-(H3-CH1) and VL3-(L3-CL) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; or (e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 210 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 211 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 212 and two light chains each comprising the amino acid sequence of SEQ ID NO: 213; NO: 212 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vi) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 213 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 225 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; viiii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 101 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; or ix) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 123 and two light chains, each containing an amino acid sequence of SEQ ID NO: 121 or 206. In some embodiments, the second target antigen is TSLP (i.e., the two antibody portions are "anti-TSLP Fab1" and "anti-TSLP Fab2"). In some embodiments, the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體包含兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)，以及第二抗體部分，其為特異性地結合至第二標靶抗原的全長抗體；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-第二抗體部分之第一重鏈的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-第二抗體部分之第二重鏈的第三多肽；iv)含有N’至C’：VL2-(L2-CL)的第四多肽；v)含有第二抗體部分之第一輕鏈的第五多肽；及vi)含有第二抗體部分之第二輕鏈的第六多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；或(e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該第二標靶抗原為TSLP (亦即，該第二抗體部分為「抗TSLP全長抗體」)。在一些實施例中，該抗TSLP全長抗體包含兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈。In some embodiments according to any of the above multispecific constructs, the multispecific construct comprises two anti-IL-13 antibody portions, which are Fab ("anti-IL-13 Fab1" and "anti-IL-13 Fab2")), and a second antibody portion that is a full-length antibody that specifically binds to a second target antigen; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-a first heavy chain of the second antibody portion; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optionally a linker-a second heavy chain of the second antibody portion; iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); v) a fifth polypeptide comprising the first light chain of the second antibody portion; and vi) a sixth polypeptide comprising the second light chain of the second antibody portion; and wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-IL-13 Fab2. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) (d) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; or (e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the second target antigen is TSLP (i.e., the second antibody portion is an "anti-TSLP full-length antibody"). In some embodiments, the anti-TSLP full-length antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 105, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 103.

在根據上述多特異性構築體之任一者的一些實施例中，該多特異性構築體包含該第一抗體部分，其為抗IL-13全長抗體，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原的Fab (「Fab1」及「Fab2」)；其中該多特異性構築體包含：i)含有N’至C’：VL3-(L3-CL)的第一多肽；ii)含有N’至C’：VH3-(H3-CH1)-視情況的連接子-抗IL-13全長抗體之第一重鏈(H1)的第二多肽；iii)含有N’至C’：VH4-(H4-CH1)-視情況的連接子-抗IL-13全長抗體之第二重鏈(H2)的第三多肽；iv)含有N’至C’：VL4-(L4-CL)的第四多肽；v)含有抗IL-13全長抗體之第一輕鏈(L1)的第五多肽；及vi)含有抗IL-13全長抗體之第二輕鏈(L2)的第六多肽；且其中VL3-(L3-CL)及VH3-(H3-CH1)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；或(e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或ix)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈。在一些實施例中，該第二標靶抗原為TSLP (亦即，該兩個抗體部分為「抗TSLP Fab1」及「抗TSLP Fab2」)。在一些實施例中，該抗TSLP Fab1及該抗TSLP Fab2各自包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。In some embodiments according to any of the above multispecific constructs, the multispecific construct comprises the first antibody portion, which is an anti-IL-13 full-length antibody, and two second antibody portions, which are Fabs that specifically bind to a second target antigen ("Fab1" and "Fab2"); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL3-(L3-CL); ii) a second polypeptide comprising N' to C': VH3-(H3-CH1)-optional linker-first heavy chain (H1) of an anti-IL-13 full-length antibody; iii) a second polypeptide comprising N' to C': VH4-(H4-CH1)-optional linker-second heavy chain ( H2); iv) a fourth polypeptide containing N' to C': VL4-(L4-CL); v) a fifth polypeptide containing the first light chain (L1) of the anti-IL-13 full-length antibody; and vi) a sixth polypeptide containing the second light chain (L2) of the anti-IL-13 full-length antibody; and wherein VL3-(L3-CL) and VH3-(H3-CH1) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; or (e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 210 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 211 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 212 and two light chains each comprising the amino acid sequence of SEQ ID NO: 213; NO: 212 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vi) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 213 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 225 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; viiii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 101 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; or ix) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 123 and two light chains, each containing an amino acid sequence of SEQ ID NO: 121 or 206. In some embodiments, the second target antigen is TSLP (i.e., the two antibody portions are "anti-TSLP Fab1" and "anti-TSLP Fab2"). In some embodiments, the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103.

亦提供包含本文所述之多特異性構築體之任一者的醫藥組成物及可選地醫藥上可接受之載體。Also provided are pharmaceutical compositions comprising any of the multispecific constructs described herein and, optionally, a pharmaceutically acceptable carrier.

亦提供編碼本文所述之多特異性構築體之任一者的分離核酸、包含此類核酸的載體及包含此類核酸或載體的宿主細胞。Also provided are isolated nucleic acids encoding any of the multispecific constructs described herein, vectors comprising such nucleic acids, and host cells comprising such nucleic acids or vectors.

亦提供治療個體之發炎性疾病的方法，其包含向該個體投予有效量之本文所述之多特異性構築體之任一者或本文所述之醫藥組成物之任一者。在一些實施例中，其中該發炎性疾病為氣喘、異位性皮膚炎或慢性阻塞性肺病(hronic obstructive pulmonary disease，COPD)。在一些實施例中，該個體為人類。Also provided are methods for treating an inflammatory disease in a subject, comprising administering to the subject an effective amount of any of the multispecific constructs described herein or any of the pharmaceutical compositions described herein. In some embodiments, the inflammatory disease is asthma, atopic dermatitis, or chronic obstructive pulmonary disease (COPD). In some embodiments, the subject is human.

亦提供產生本文所述之多特異性構築體之任一者的方法，其包含i)在適合表現該多特異性構築體的條件下培養含有上述之分離的核酸或載體之任一者的宿主細胞或上述宿主細胞之任一者；及ii)獲得該表現的多特異性構築體。Also provided are methods for producing any of the multispecificity constructs described herein, comprising i) culturing a host cell containing any of the isolated nucleic acids or vectors described above, or any of the host cells described above, under conditions suitable for expression of the multispecificity construct; and ii) obtaining the expressed multispecificity construct.

在另一態樣中，本發明提供分離的抗體構築體(抗IL-13抗體構築體)，其包含特異性地結合IL-13的抗體部分(「抗IL-13抗體部分」)，其中該抗IL-13抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，且其中：(1)該VH包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3。In another aspect, the present invention provides an isolated antibody construct (anti-IL-13 antibody construct) comprising an antibody portion that specifically binds IL-13 ("anti-IL-13 antibody portion"), wherein the anti-IL-13 antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), and wherein: (1) the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: CDR-L3 of the amino acid sequence of NO:72.

在根據上述之抗IL-13抗體構築體之任一者的一些實施例中，(1)該VH含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80%序列同一性的變體，且該VL含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80%序列同一性的變體。在一些實施例中，該VH含有SEQ ID NO:65之胺基酸序列，且該VL含有SEQ ID NO:69之胺基酸序列。In some embodiments according to any of the above anti-IL-13 antibody constructs, (1) the VH comprises the amino acid sequence of SEQ ID NO: 65 or a variant thereof having at least about 80% sequence identity with SEQ ID NO: 65, and the VL comprises the amino acid sequence of SEQ ID NO: 69 or a variant thereof having at least about 80% sequence identity with SEQ ID NO: 69. In some embodiments, the VH comprises the amino acid sequence of SEQ ID NO: 65, and the VL comprises the amino acid sequence of SEQ ID NO: 69.

在根據上述之抗IL-13抗體構築體之任一者的一些實施例中，該抗IL-13抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、雙抗體及scFv。In some embodiments according to any of the above anti-IL-13 antibody constructs, the anti-IL-13 antibody portion is selected from the group consisting of a full-length antibody, Fab, Fab', F(ab')2, a diabody, and a scFv.

在根據上述之抗IL-13抗體構築體之任一者的一些實施例中，該抗IL-13抗體部分為scFv (「抗IL-13 scFv」)。在一些實施例中，該抗IL-13 scFv含有SEQ ID NO:108之胺基酸序列。In some embodiments of any of the anti-IL-13 antibody constructs described above, the anti-IL-13 antibody portion is a scFv ("anti-IL-13 scFv"). In some embodiments, the anti-IL-13 scFv comprises the amino acid sequence of SEQ ID NO: 108.

在根據上述之抗IL-13抗體構築體之任一者的一些實施例中，該抗IL-13抗體部分為Fab (「抗IL-13 Fab」)。在一些實施例中，該抗IL-13 Fab包含含有SEQ ID NO:124之胺基酸序列的第一多肽及含有SEQ ID NO:102或197之胺基酸序列的第二多肽。In some embodiments of any of the anti-IL-13 antibody constructs described above, the anti-IL-13 antibody portion is a Fab ("anti-IL-13 Fab"). In some embodiments, the anti-IL-13 Fab comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197.

在根據上述之抗IL-13抗體構築體之任一者的一些實施例中，該抗IL-13抗體部分為全長抗體(「抗IL-13全長抗體」)。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；ix)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或x)含有SEQ ID NO:130之胺基酸序列的第一重鏈、含有SEQ ID NO:131之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。In some embodiments according to any of the above anti-IL-13 antibody constructs, the anti-IL-13 antibody portion is a full-length antibody ("anti-IL-13 full-length antibody"). In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 101 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 123 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 101 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; NO: 210 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207; vi) two heavy chains each containing the amino acid sequence of SEQ ID NO: 211 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206; vii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 212 and two light chains each containing the amino acid sequence of SEQ ID NO: 120 or 208; viii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 213 and two light chains each containing the amino acid sequence of SEQ ID NO: 120 or 208; ix) two heavy chains each containing the amino acid sequence of SEQ ID NO: 225 and two light chains each containing the amino acid sequence of SEQ ID NO: or x) a first heavy chain comprising the amino acid sequence of SEQ ID NO: 130, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 131, and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197.

在根據上述之抗IL-13抗體構築體之任一者的一些實施例中，該分離的抗IL-13抗體構築體係單特異性。在一些實施例中，該分離的抗IL-13抗體構築體係多特異性。In some embodiments of any of the above anti-IL-13 antibody constructs, the isolated anti-IL-13 antibody construct is monospecific. In some embodiments, the isolated anti-IL-13 antibody construct is multispecific.

在根據上述之抗IL-13抗體構築體之任一者的一些實施例中，該分離的抗IL-13抗體構築體進一步包含特異性地結合至第二抗原的第二抗體部分。在一些實施例中，該抗IL-13抗體部分及該第二抗體部分係經由連接子彼此融合。在一些實施例中，該連接子含有GG及SEQ ID NO:98-99之任一者的胺基酸序列。In some embodiments of any of the above anti-IL-13 antibody constructs, the isolated anti-IL-13 antibody construct further comprises a second antibody portion that specifically binds to a second antigen. In some embodiments, the anti-IL-13 antibody portion and the second antibody portion are fused to each other via a linker. In some embodiments, the linker comprises GG and the amino acid sequence of any one of SEQ ID NOs: 98-99.

亦提供包含本文所述之分離的抗IL-13抗體構築體之任一者的醫藥組成物及可選地醫藥上可接受之載體。Also provided are pharmaceutical compositions comprising any of the isolated anti-IL-13 antibody constructs described herein, and optionally a pharmaceutically acceptable carrier.

亦提供編碼本文所述之分離的抗IL-13抗體構築體之任一者的分離核酸、包含此類核酸的載體及包含此類核酸或載體的宿主細胞。Also provided are isolated nucleic acids encoding any of the isolated anti-IL-13 antibody constructs described herein, vectors comprising such nucleic acids, and host cells comprising such nucleic acids or vectors.

亦提供治療個體之發炎性疾病的方法，其包含向該個體投予有效量之本文所述之分離的抗IL-13抗體構築體之任一者或本文所述之醫藥組成物之任一者。在一些實施例中，該發炎性疾病為氣喘、異位性皮膚炎或COPD。在一些實施例中，該個體為人類。Also provided are methods for treating an inflammatory disease in a subject, comprising administering to the subject an effective amount of any of the isolated anti-IL-13 antibody constructs described herein or any of the pharmaceutical compositions described herein. In some embodiments, the inflammatory disease is asthma, atopic dermatitis, or COPD. In some embodiments, the subject is human.

亦提供產生本文所述之分離的抗IL-13抗體構築體之任一者的方法，其包含：i)在適合表現該抗IL-13抗體構築體的條件下培養含有上述之分離的核酸或載體之任一者的宿主細胞或上述宿主細胞之任一者；及ii)獲得該表現的抗IL-13抗體構築體。Also provided are methods for producing any of the isolated anti-IL-13 antibody constructs described herein, comprising: i) culturing a host cell containing any of the isolated nucleic acids or vectors described above, or any of the host cells described above, under conditions suitable for expression of the anti-IL-13 antibody construct; and ii) obtaining the expressed anti-IL-13 antibody construct.

本發明之此等及其他態樣及優點將由後續實施方式及所附申請專利範圍而變得顯而易見。應理解，可組合本文所述之各種實施例的一者、一些或所有性質以形成本發明之其他實施例。These and other aspects and advantages of the present invention will become apparent from the subsequent implementation and the appended claims. It should be understood that one, some, or all of the properties of the various embodiments described herein may be combined to form other embodiments of the present invention.

本文提及的所有出版品、專利、專利申請案及公開的專利申請案的揭露內容皆通過引用整體併入本文中。The disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are incorporated by reference in their entirety.

本發明提供一種多特異性構築體(諸如雙特異性構築體)，其包含特異性地結合至IL-13的抗IL-13抗體部分及特異性地結合至第二標靶抗原(例如，TSLP)的第二抗體部分。在一些實施例中，該多特異性構築體包含一個抗IL-13抗體部分。在一些實施例中，該多特異性構築體包含二或多個抗IL-13抗體部分。在一些實施例中，該多特異性構築體包含一個特異性地結合至第二標靶抗原或標靶表位的抗體部分。在一些實施例中，該多特異性構築體包含二或多個特異性地結合至一或多個其他標靶抗原或標靶表位的抗體部分。在一些實施例中，該第二抗原為由免疫細胞產生的蛋白質。在一些實施例中，該第二抗原為TSLP。在另一態樣中，本發明亦提供新穎的抗IL-13抗體構築體。The present invention provides a multispecific construct (e.g., a bispecific construct) comprising an anti-IL-13 antibody portion that specifically binds to IL-13 and a second antibody portion that specifically binds to a second target antigen (e.g., TSLP). In some embodiments, the multispecific construct comprises one anti-IL-13 antibody portion. In some embodiments, the multispecific construct comprises two or more anti-IL-13 antibody portions. In some embodiments, the multispecific construct comprises one antibody portion that specifically binds to a second target antigen or target epitope. In some embodiments, the multispecific construct comprises two or more antibody portions that specifically bind to one or more additional target antigens or target epitopes. In some embodiments, the second antigen is a protein produced by an immune cell. In some embodiments, the second antigen is TSLP. In another aspect, the present invention also provides novel anti-IL-13 antibody constructs.

在廣泛研究之後，本申請案之發明人發現，本文所述之多特異性構築體(例如，抗IL-13多特異性構築體)與其他多特異性蛋白質相比具有幾個意想不到的優勢。首先，與目前可用的抗IL-13抗體或其多特異性構築體相比，本文所述之多特異性構築體表現出與IL-13更強的結合及提高的效力。其次，該多特異性構築體(例如，抗IL-13多特異性構築體)與人類及食蟹獼猴IL-13皆具有交叉反應性，其可促進將食蟹獼猴毒性及功效研究的結果推斷至人類臨床研究。第三，本文所述之某些多特異性構築體顯示在體內具有驚人的穩定性(例如，在投予小鼠及食蟹獼猴時)，使其等特別適合以高濃度調配。第四，本文所述之某些多特異性構築體的體內半衰期顯示比其他對照多特異性抗體長得多(例如，該半衰期為對照多特異性抗體之半衰期的至少約1.5倍、2倍、4倍、5倍或更多)。After extensive research, the inventors of this application discovered that the multispecific constructs described herein (e.g., anti-IL-13 multispecific constructs) possess several unexpected advantages over other multispecific proteins. First, compared to currently available anti-IL-13 antibodies or multispecific constructs thereof, the multispecific constructs described herein exhibit stronger binding to IL-13 and improved potency. Second, the multispecific constructs (e.g., anti-IL-13 multispecific constructs) exhibit cross-reactivity with both human and cynomolgus macaque IL-13, which facilitates the extrapolation of results from cynomolgus macaque toxicity and efficacy studies to human clinical studies. Third, certain multispecific constructs described herein exhibit remarkable stability in vivo (e.g., when administered to mice and cynomolgus macaques), making them particularly suitable for high-concentration formulation. Fourth, certain multispecific constructs described herein exhibit significantly longer in vivo half-lives than other control multispecific antibodies (e.g., half-lives that are at least about 1.5-fold, 2-fold, 4-fold, 5-fold, or more than the half-life of the control multispecific antibody).

亦提供包含本文所述之多特異性構築體或抗IL-13抗體構築體之任一者的醫藥組成物及套組，以及其用於治療發炎性疾病(諸如氣喘、異位性皮膚炎或COPD)的使用方法。I.定義Also provided are pharmaceutical compositions and kits comprising any of the multispecific constructs or anti-IL-13 antibody constructs described herein, and methods of use thereof for treating inflammatory diseases such as asthma, atopic dermatitis, or COPD.I.Definitions

如本文所用，術語「治療」意指經設計以改變所治療之個體或細胞在臨床病理學之病程期間的天然過程的臨床介入。所需治療效果包括降低疾病進展速率、改善或緩和疾病狀態及緩解或改善預後。舉例而言，若與發炎性疾病相關之一或多種症狀得到減輕或消除，包括但不限於降低局部或全身發炎、減少由疾病引起之症狀、提高罹患疾病者之生活品質、降低治療疾病所需之其他藥物的劑量等，則個體為成功「治療的」。As used herein, the term "treatment" refers to clinical interventions designed to alter the natural course of the treated individual or cells during the course of a clinical pathology. Desirable therapeutic effects include a reduction in the rate of disease progression, amelioration or alleviation of the disease state, and palliation or improved prognosis. For example, a subject is successfully "treated" if one or more symptoms associated with an inflammatory disease are reduced or eliminated, including but not limited to a reduction in local or systemic inflammation, a decrease in symptoms caused by the disease, an improvement in the quality of life of the subject suffering from the disease, a reduction in the dosage of other medications required to treat the disease, etc.

如本文所用，「有效量」意指有效治療受試者(諸如個體，例如人類)之疾病或病症的藥劑或藥物的量。在發炎性疾病的情況下，該藥劑的有效量可減少活性免疫細胞的數量；減少促發炎性細胞激素的量；局部性地及/或全身性地抑制(亦即，在一定程度上減緩，較佳地停止)發炎性免疫細胞活性；及/或在一定程度上緩解與發炎性疾病相關的一或多種症狀。如在臨床背景中所理解的，藥物、化合物或藥物組成物之有效量可與或不與另一藥物、化合物或醫藥組成物聯合而達成。因此，在投予一或多種治療劑之上下文中可考慮「有效量」，且若單一藥劑與一或多種其他藥劑聯合而可達到或已達到所需結果，則該單一藥劑可視為以有效量給出。As used herein, an "effective amount" refers to an amount of an agent or drug that is effective in treating a disease or condition in a subject (e.g., an individual, such as a human). In the case of an inflammatory disease, an effective amount of the agent can reduce the number of active immune cells; reduce the amount of pro-inflammatory cytokines; inhibit (i.e., reduce to some extent, preferably stop) inflammatory immune cell activity locally and/or systemically; and/or alleviate to some extent one or more symptoms associated with the inflammatory disease. As understood in the clinical context, an effective amount of a drug, compound, or pharmaceutical composition can be achieved with or without combination with another drug, compound, or pharmaceutical composition. Thus, an "effective amount" may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if it can achieve or has achieved the desired result in combination with one or more other agents.

如本文所用，「個體」或「受試者」意指哺乳動物，包括但不限於人類、牛、馬、貓、犬、囓齒動物或靈長類動物。在一些實施例中，該個體為人類。As used herein, "individual" or "subject" refers to mammals, including but not limited to humans, cows, horses, cats, dogs, rodents, or primates. In some embodiments, the individual is a human.

術語「抗體」係以最廣泛意義使用且具體地涵蓋單株抗體(包括全長單株抗體)、多特異性抗體(例如，雙特異性抗體)及抗體片段，只要其等展現出所需生物活性或功能。如本文所用，術語「免疫球蛋白(Ig)」及「抗體」可互換使用。The term "antibody" is used in the broadest sense and specifically encompasses monoclonal antibodies (including full-length monoclonal antibodies), multispecific antibodies (e.g., bispecific antibodies), and antibody fragments, so long as they exhibit the desired biological activity or function. As used herein, the terms "immunoglobulin (Ig)" and "antibody" are used interchangeably.

如本文所用，術語「全長抗體」意指呈其實質上完整形式、不為如下文所定義之抗體片段的抗體。特別地，該等術語意指具有含Fc區之重鏈的抗體。全長抗體通常為約150,000道耳吞的異四聚化醣蛋白，其由兩條相同的輕(L)鏈及兩條相同的重(H)鏈組成。As used herein, the term "full-length antibody" refers to an antibody in its substantially intact form, not as an antibody fragment as defined below. In particular, the term refers to an antibody having a heavy chain containing an Fc region. Full-length antibodies are typically heterotetrameric glycoproteins of approximately 150,000 dU, composed of two identical light (L) chains and two identical heavy (H) chains.

術語「恆定域」意指具有相對於含有抗原結合位點之免疫球蛋白之其他部分(可變域)更保留之胺基酸序列的免疫球蛋白分子之部分。恆定域含有重鏈之CH1、CH2及CH3域(統稱為CH)及輕鏈之CHL (或CL)域。The term "constant domain" refers to the portion of an immunoglobulin molecule that has a more conserved amino acid sequence than the rest of the immunoglobulin (variable domain) that contains the antigen-binding site. The constant domain contains the heavy chainCH1 ,CH2 , and CH3 domains (collectively referred to as CH) and the light chainCHL (or CL) domain.

抗體之「可變區」或「可變域」意指抗體之重鏈或輕鏈的胺基末端域。重鏈之可變域可稱為「VH」。輕鏈之可變域可稱為「VL」。此等域通常為抗體之最可變部分且含有抗原結合位點。The term "variable region" or "variable domain" of an antibody refers to the amino-terminal domain of either the heavy or light chain of an antibody. The variable domain of the heavy chain may be referred to as "VH." The variable domain of the light chain may be referred to as "VL." These domains are generally the most variable portion of the antibody and contain the antigen-binding site.

術語「可變」意指以下事實：可變域之某些部分在抗體當中在序列方面廣泛地不同，且用於各特定抗體對於其特定抗原之結合及特異性。然而，可變性並非均勻分佈於抗體的整個可變域中。其集中在輕鏈及重鏈可變域中之三個稱作高度可變區(HVR，亦稱為CDR)的區段中。可變域中保留性較高之部分稱為框架區(FR)。天然重鏈及輕鏈之可變域各自包含四個 FR，主要採用 β-折疊構型，藉由三個HVR連接，其形成連接β-折疊結構之環並在一些情況下形成β-折疊結構之一部分。各鏈中之HVR係藉由FR緊密地結合在一起，且與另一鏈之HVR一起，有助於形成抗體之抗原結合位點(參見Kabat等人，Sequences of Proteins of Immunological Interest，第五版，National Institute of Health，Bethesda，Md. (1991))。恆定域不直接參與抗體與抗原的結合，但展現出各種效應功能，諸如抗體依賴性細胞毒性中抗體的參與。本文所述之恆定域的殘基係根據EU編號。The term "variable" refers to the fact that certain portions of the variable domain differ extensively in sequence among antibodies and contribute to the binding and specificity of each particular antibody for its particular antigen. However, variability is not evenly distributed throughout the variable domain of an antibody. It is concentrated in three segments called highly variable regions (HVRs, also called CDRs) in both the light and heavy chain variable domains. The more conserved portions of the variable domains are called framework regions (FRs). The native heavy and light chain variable domains each contain four FRs, primarily adopting a β-sheet configuration, connected by three HVRs, which form loops connecting and, in some cases, forming part of the β-sheet structure. The HVRs in each chain are tightly bound together by the FRs and, together with the HVRs of the other chain, contribute to the formation of the antibody antigen-binding site (see Kabat et al., Sequences of Proteins of Immunological Interest, 5th ed., National Institute of Health, Bethesda, Md. (1991)). The constant domain is not directly involved in antibody-antigen binding but exhibits various effector functions, such as participation of antibodies in antibody-dependent cellular cytotoxicity. The constant domain residues described herein are based on the EU numbering.

如本文所用，術語「CDR」或「互補決定區」意指重鏈及輕鏈多肽之可變區內發現的非連續抗原結合位點。在文獻Kabat等人，J. Biol. Chem. 252:6609-6616 (1977)；Kabat等人，U.S. Dept. of Health and Human Services, “Sequences of proteins of immunological interest” (1991)；Chothia等人，J. Mol. Biol. 196:901-917 (1987)；Al-Lazikani B.等人，J. Mol. Biol., 273: 927-948 (1997)；MacCallum等人，J. Mol. Biol. 262:732-745 (1996)；Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008)；Lefranc M.P.等人，Dev. Comp. Immunol., 27: 55-77 (2003)；及Honegger and Plückthun, J. Mol. Biol., 309:657-670 (2001)中已描述此等特定區域，其中當彼此之間互相比較時，此等定義包括胺基酸殘基的重合或子集。儘管如此，應用任何一種定義方式來指示抗體或移植抗體或其變體之CDR，皆包括在本文所定義及使用的術語範疇內。表A中列出由上述引用的各篇參考文獻所定義的CDR所包括的胺基酸殘基，以作為比較。CDR預測的演算法及結合介面在本領域為已知的，包括例如：Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008)；Ehrenmann F.等人，Nucleic Acids Res., 38: D301-D307 (2010)；及Adolf-Bryfogle J.等人，Nucleic Acids Res., 43: D432-D438 (2015)。本段中所引用的參考文獻的內容以其整體引用併入本文中，以用於本申請案及可能包含在本文的一或多個請求項中。本文提供的CDR的胺基酸殘基係基於Kabat進行描述。表A. CDR定義Kabat1Chothia2MacCallum3IMGT4Aho5VH CDR131-3526-3230-3527-3825-40VH CDR250-6553-5547-5856-6558-77VH CDR395-10296-10193-101105-117109-137VL CDR124-3426-3230-3627-3825-40VL CDR250-5650-5246-5556-6558-77VL CDR389-9791-9689-96105-117109-1371殘基編號按照上述Kabat等人的命名法2殘基編號按照上述Chothia等人的命名法3殘基編號按照上述MacCallum等人的命名法4殘基編號按照上述Lefranc等人的命名法5殘基編號按照上述Honegger及Plückthun的命名法As used herein, the term "CDR" or "complementary determining region" refers to the non-contiguous antigen binding sites found within the variable regions of heavy and light chain polypeptides. In the literature, Kabat et al., J. Biol. Chem. 252:6609-6616 (1977); Kabat et al., US Dept. of Health and Human Services, “Sequences of proteins of immunological interest” (1991); Chothia et al., J. Mol. Biol. 196:901-917 (1987); Al-Lazikani B. et al., J. Mol. Biol., 273: 927-948 (1997); MacCallum et al., J. Mol. Biol. 262:732-745 (1996); Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008); Lefranc MP et al., Dev. Comp. Immunol., 27: 55-77 (2003); and Honegger et al. These specific regions are described in [15] and Plückthun, J. Mol. Biol., 309:657-670 (2001), wherein these definitions include overlap or subsets of amino acid residues when compared to one another. Nevertheless, any definition used to designate the CDRs of an antibody or grafted antibody or variant thereof is included within the scope of the terms defined and used herein.TableA lists the amino acid residues included in the CDRs defined in the above-cited references for comparison. Algorithms and binding interfaces for CDR prediction are known in the art, including, for example, Abhinandan and Martin, Mol. Immunol., 45: 3832-3839 (2008); Ehrenmann F. et al., Nucleic Acids Res., 38: D301-D307 (2010); and Adolf-Bryfogle J. et al., Nucleic Acids Res., 43: D432-D438 (2015). The contents of the references cited in this paragraph are incorporated herein by reference in their entirety for use in this application and in one or more claims that may be included herein. The amino acid residues of the CDRs provided herein are described based on Kabat.TableA. CDRDefinitionsKabat1Chothia2MacCallum3IMGT4Aho5 VH CDR1 31-35 26-32 30-35 27-38 25-40 VH CDR2 50-65 53-55 47-58 56-65 58-77 VH CDR3 95-102 96-101 93-101 105-117 109-137 VL CDR1 24-34 26-32 30-36 27-38 25-40 VL CDR2 50-56 50-52 46-55 56-65 58-77 VL CDR3 89-97 91-96 89-96 105-117 109-1371 Residue numbering according to the nomenclature of Kabat et al.2 Residue numbering according to the nomenclature of Chothia et al.3 Residue numbering according to the nomenclature of MacCallum et al.4 Residue numbering according to the nomenclature of Lefranc et al.5 Residue numbering according to the nomenclature of Honegger and Plückthun

任何哺乳動物物種之抗體(免疫球蛋白)的「輕鏈」皆可根據其恆定域之胺基酸序列被歸類為兩種截然不同的類型(稱為卡帕(「κ」)及拉目達(「λ」))之一。The "light chains" of antibodies (immunoglobulins) from any mammalian species can be classified into one of two distinct types, called kappa ("κ") and lambda ("λ"), based on the amino acid sequence of their homeodomains.

如本文所用，術語IgG 「同型」或「子類」意指由其恆定區之化學及抗原特徵所定義之免疫球蛋白子類之任一者。As used herein, the term IgG "isotype" or "subclass" refers to any of the immunoglobulin subclasses defined by the chemical and antigenic characteristics of its constant regions.

根據其重鏈恆定域之胺基酸序列，抗體(免疫球蛋白)可歸類為不同的類別。有五大類免疫球蛋白：IgA、IgD、IgE、IgG及IgM，且彼等中之數種可進一步分為子類(同型)，例如IgG1、IgG2、IgG3、IgG4、IgA1及IgA2。對應於不同類別之免疫球蛋白的重鏈恆定域分別稱為 α、δ、ɛ、γ 及 μ。不同類別的免疫球蛋白之次單位結構及三維構型為熟習的，且一般描述於例如Abbas等人之Cellular and Mol. Immunology，第 4 版(W.B.Saunders, Co., 2000)。抗體可為較大融合分子之一部分，其藉由抗體與一或多種其他蛋白質或肽的共價或非共價締合形成。Antibodies (immunoglobulins) can be classified into different classes based on the amino acid sequences of their heavy-chain constant domains. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these can be further divided into subclasses (isotypes), such as IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The heavy-chain constant domains corresponding to the different classes of immunoglobulins are called α, δ, ɛ, γ, and μ, respectively. The subunit structures and three-dimensional configurations of the different classes of immunoglobulins are well known and generally described in, for example, Abbas et al., Cellular and Mol. Immunology, 4th ed. (W.B. Saunders, Co., 2000). An antibody may be part of a larger fusion molecule, formed by covalent or non-covalent association of the antibody with one or more other proteins or peptides.

「抗體片段」包含完整抗體之一部分，較佳地包含其抗原結合區。在一些實施例中，本文所述之抗體片段為抗原結合片段。抗體片段或抗原結合片段之實例包括Fab、Fab'、F(ab')2及Fv片段(諸如單鏈可變片段，scFv)；雙功能抗體；線性抗體；單鏈抗體分子；及由抗體片段形成之多特異性抗體。An "antibody fragment" comprises a portion of an intact antibody, preferably comprising its antigen-binding region. In some embodiments, the antibody fragments described herein are antigen-binding fragments. Examples of antibody fragments or antigen-binding fragments include Fab, Fab', F(ab')2 , and Fv fragments (e.g., single-chain variable fragments, scFv); bifunctional antibodies; linear antibodies; single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.

抗體之番木瓜蛋白酶(papain)消化產生兩個相同的抗原結合片段，稱為「Fab」片段，各自具有單一抗原結合位點，以及殘餘「Fc」片段，其名稱反映出其容易結晶的能力。胃蛋白酶(pepsin)處理產生F(ab')2片段，其具有兩個抗原組合位點且仍能夠與抗原交聯。Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment, whose name reflects its ability to readily crystallize. Pepsin treatment produces the F(ab')2 fragment, which has two antigen-binding sites and is still capable of cross-linking to the antigen.

「Fv」為含有整個抗原結合位點的最小抗體片段。在一實施例中，雙鏈Fv物種由一個重鏈及一個輕鏈可變域以緊密、非共價結合之二聚體組成。在單鏈Fv (scFv)物種中，一個重鏈及一個輕鏈可變域可藉由撓性肽連接子共價連接，使得輕鏈及重鏈可結合於類似於雙鏈Fv物種中之結構的「二聚體」結構中。在此組態中，各可變域之三個HVR相互作用以界定VH-VL二聚體表面上之抗原結合位點。六個HVR共同地賦予抗體以抗原結合特異性。然而，即使單一可變域 (或僅包含對抗原具有特異性之三個HVR的Fv的一半)亦具有辨識及結合抗原的能力，儘管其親和力低於整個結合位點。"Fv" is the smallest antibody fragment that contains an entire antigen-binding site. In one embodiment, a two-chain Fv species consists of a dimer of one heavy-chain and one light-chain variable domain in tight, non-covalent association. In a single-chain Fv (scFv) species, one heavy-chain and one light-chain variable domain can be covalently linked by a flexible peptide linker, allowing the light and heavy chains to associate in a "dimer" structure similar to that in two-chain Fv species. In this configuration, the three HVRs of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six HVRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three HVRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

Fab片段具有兩條多肽鏈，其含有重鏈及輕鏈可變域(VH、VL)，且亦含有輕鏈(CL)之恆定域及重鏈之第一恆定域 (CH1)。Fab’片段與Fab片段不同之處在於，在重鏈CH1域之羧基端添加幾個殘基，包括來自抗體鉸鏈區之一或多個半胱胺酸。Fab’-SH為其中恆定域之半胱胺酸殘基攜有游離硫醇基之Fab’在本文中的名稱。F(ab’)2抗體片段最初係以其間具有鉸鏈半胱胺酸之Fab’片段對形式產生。抗體片段之其他化學耦聯亦為已知的。Fab fragments have two polypeptide chains, containing heavy and light chain variable domains (VH, VL), as well as the homeostatic domain of the light chain (CL) and the first homeostatic domain of the heavy chain (CH1 ). Fab' fragments differ from Fab fragments by the addition of several residues to the carboxyl terminus of the heavy chainCH1 domain, including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab' fragments in which the cysteine residues of the homeostatic domains bear free thiol groups. F(ab')2 antibody fragments were originally generated as pairs of Fab' fragments with hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

「單鏈Fv」或「scFv」抗體片段包含抗體之VH域及VL域，其中此等域存在於單一多肽鏈中。一般而言，scFv多肽在VH域及VL域之間進一步包含多肽連接子，其使得scFv能夠形成用於抗原結合之所需結構。關於scFv之回顧，參見例如Plückthun，The Pharmacology of Monoclonal Antibodies，Springer Berlin Heidelberg，1994. 269-315。"Single-chain Fv" or "scFv" antibody fragments comprise the VH and VL domains of an antibody, wherein these domains are present in a single polypeptide chain. Generally, scFv polypeptides further comprise a polypeptide linker between the VH and VL domains, which enables the scFv to form the desired structure for antigen binding. For a review of scFv, see, for example, Plückthun, The Pharmacology of Monoclonal Antibodies, Springer Berlin Heidelberg, 1994, 269-315.

「Fc」片段包含兩條重鏈的羧基端部分由雙硫鍵緊密連接在一起。抗體之效應功能由Fc區中的序列決定，該區域亦為某些類型細胞上發現的Fc受體(FcR)所辨識的區域。The "Fc" fragment consists of the carboxyl-terminal portions of two heavy chains tightly linked by disulfide bonds. The effector function of an antibody is determined by the sequence in the Fc region, which is also the region recognized by Fc receptors (FcRs) found on certain cell types.

如本文所用，術語「單株抗體」意指獲自實質上同質之抗體群體的抗體，例如構成該群體之個別抗體為相同的，除了可少量存在之可能的突變，例如天然存在之突變。因此，修飾語「單株」表明抗體不為不同抗體之混合物的特徵。在一些實施例中，此類單株抗體通常包括含有結合標靶之多肽序列的抗體，其中該標靶結合多肽序列係藉由包括自複數個多肽序列選擇單一標靶結合多肽序列之方法獲得。舉例而言，選擇方法可為從複數個殖株(諸如一組融合瘤殖株、噬菌體殖株或重組DNA殖株)中選擇獨特的殖株。應理解，所選標靶結合序列可經進一步改變以例如改善針對標靶之親和力、人源化標靶結合序列、改善其於細胞培養物中之產生、降低其活體內免疫原性、產生多特異性抗體等，且包含經改變標靶結合序列之抗體亦為本發明之單株抗體。與通常包括針對不同決定位(表位)之不同抗體的多株抗體製備物相反，單株抗體製備物之各個單株抗體係針對於抗原上的單一決定位。除其特異性以外，單株抗體製備物亦為有利的，係因其通常未被其他免疫球蛋白污染。As used herein, the term "monoclonal antibody" means an antibody obtained from a substantially homogeneous population of antibodies, e.g., the individual antibodies comprising the population are identical except for possible mutations, such as naturally occurring mutations, that may be present in small amounts. Thus, the modifier "monoclonal" indicates the characteristic that the antibody is not a mixture of different antibodies. In some embodiments, such monoclonal antibodies generally include antibodies comprising a polypeptide sequence that binds a target, wherein the target-binding polypeptide sequence is obtained by a method that includes selecting a single target-binding polypeptide sequence from a plurality of polypeptide sequences. For example, the selection method can be selecting a unique strain from a plurality of strains, such as a set of fusion tumor strains, phage strains, or recombinant DNA strains. It should be understood that the selected target binding sequence can be further altered to, for example, improve affinity for the target, humanize the target binding sequence, improve its production in cell culture, reduce its in vivo immunogenicity, generate multispecific antibodies, etc., and that antibodies comprising altered target binding sequences are also monoclonal antibodies of the present invention. In contrast to polyclonal antibody preparations, which typically include different antibodies directed against different determinants (epitopes), each monoclonal antibody in a monoclonal antibody preparation is directed against a single determinant on the antigen. In addition to their specificity, monoclonal antibody preparations are also advantageous because they are generally free of contamination by other immunoglobulins.

修飾詞「單株」表明抗體之特徵係獲自實質上同源之抗體群體，且不應解釋為需要藉由任何特定方法產生該抗體。舉例而言，待根據本發明使用的單株抗體可藉由多種技術製備，包括例如：融合瘤方法(例如，Kohler及Milstein，Nature 256:495-97 (1975)；Hongo等人，Hybridoma 14 (3): 253-260 (1995)，Harlow等人，Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press，第2版，1988)；Hammerling等人，Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier，N.Y.，1981))、重組DNA方法(參見例如，美國專利號4,816,567)、噬菌體展示技術(參見例如Clackson等人，Nature 352: 624-628 (1991)；Marks等人，J. Mol. Biol. 222: 581-597 (1992)；Sidhu等人，J. Mol. Biol. 338(2): 299-310 (2004)；Lee等人，J. Mol. Biol. 340(5): 1073-1093 (2004)；Fellouse，Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004)；及Lee等人，J. Immunol. Methods 284(1-2): 119-132 (2004))，以及用於在具有人類免疫球蛋白基因座或編碼人類免疫球蛋白序列的基因之一部分或全部的動物中產生人類或人類樣抗體的技術(參見例如WO 1998/24893；WO 1996/34096；WO 1996/33735；WO 1991/10741；Jakobovits等人，Proc. Natl. Acad. Sci. USA 90: 2551 (1993)；Jakobovits等人，Nature 362: 255-258 (1993)；Bruggemann等人，Year in Immunol. 7:33 (1993)；美國專利號5,545,807；5,545,806；5,569,825；5,625,126；5,633,425；及5,661,016；Marks等人，Bio/Technology 10: 779-783 (1992)；Lonberg等人，Nature 368: 856-859 (1994)；Morrison，Nature 368: 812-813 (1994)；Fishwild等人，Nature Biotechnol. 14: 845-851 (1996)；Neuberger，Nature Biotechnol. 14: 826 (1996)；及Lonberg及Huszar，Intern. Rev. Immunol. 13: 65-93 (1995))。The modifier "monoclonal" indicates that the antibody is characterized as being derived from a substantially homogeneous population of antibodies and should not be construed as requiring production of the antibody by any particular method. For example, monoclonal antibodies to be used according to the present invention can be prepared by a variety of techniques, including, for example, the hypoderma method (e.g., Kohler and Milstein, Nature 256:495-97 (1975); Hongo et al., Hybridoma 14 (3): 253-260 (1995), Harlow et al., Antibodies: A Laboratory Manual, (Cold Spring Harbor Laboratory Press, 2nd ed., 1988); Hammerling et al., Monoclonal Antibodies and T-Cell Hybridomas 563-681 (Elsevier, N.Y., 1981)), recombinant DNA methods (see, e.g., U.S. Patent No. 4,816,567), phage display technology (see, e.g., Clackson et al., Nature 352: 624-628 (1991); Marks et al., J. Mol. Biol. 222: 581-597 (1992); Sidhu et al., J. Mol. Biol. 338(2): 299-310 (2004); Lee et al., J. Mol. Biol. 340(5): 1073-1093 (2004); Fellouse, Proc. Natl. Acad. Sci. USA 101(34): 12467-12472 (2004); and Lee et al., J. Immunol. Methods 284(1-2): 119-132 (2004)), and techniques for producing human or human-like antibodies in animals that have a human immunoglobulin locus or a portion or all of a gene encoding a human immunoglobulin sequence (see, e.g., WO 1998/24893; WO 1996/34096; WO 1996/33735; WO 1991/10741; Jakobovits et al., Proc. Natl. Acad. Sci. USA 90: 2551 (1993); Jakobovits et al., Nature 362: 255-258 (1993); Bruggemann et al., Year in Immunol. 7:33 (1993); U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; and 5,661,016; Marks et al., Bio/Technology 10: 779-783 (1992); Lonberg et al., Nature 368: 856-859 (1994); Morrison, Nature 368: 812-813 (1994); Fishwild et al., Nature Biotechnol. 14: 845-851 (1996); Neuberger, Nature Biotechnol. 14: 826 (1996); and Lonberg and Huszar, Intern. Rev. Immunol. 13: 65-93 (1995)).

本文中之單株抗體具體地包括「嵌合」抗體，其中重鏈及/或輕鏈之一部分與衍生自特定物種或隸屬特定抗體類別或子類之抗體的對應序列一致或同源，而各鏈之其餘部分與衍生自另一物種或隸屬另一抗體類別或子類之抗體以及此類抗體之片段的對應序列一致或同源，只要其展現出所需之生物活性即可(參見例如，美國專利號4,816,567；及Morrison等人，Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984))。嵌合抗體包括PRIMATIZED®抗體，其中該抗體之抗原結合區係衍生自藉由例如利用感興趣之抗原使獼猴免疫而產生的抗體。Monoclonal antibodies herein specifically include "chimeric" antibodies, in which a portion of the heavy and/or light chains is identical or homologous to corresponding sequences in antibodies derived from a particular species or belonging to a particular antibody class or subclass, while the remainder of each chain is identical or homologous to corresponding sequences in antibodies derived from another species or belonging to another antibody class or subclass, as well as fragments of such antibodies, so long as they exhibit the desired biological activity (see, e.g., U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA 81:6851-6855 (1984)). Chimeric antibodies include PRIMATIZED® antibodies, in which the antigen-binding region of the antibody is derived from an antibody generated, for example, by immunizing macaques with the antigen of interest.

非人類(例如，鼠類)抗體之「人源化」形式為含有衍生自非人類免疫球蛋白之最小序列的嵌合抗體。在一實施例中，人源化抗體為人類免疫球蛋白(受體抗體)，其中來自受體HVR的殘基經來自具有所需特異性、親和力及/或容量之非人類物種(供體抗體)，諸如小鼠、大鼠、兔或非人類靈長類動物之HVR的殘基置換。在一些情況下，人類免疫球蛋白之FR殘基經相應之非人類殘基置換。此外，人源化抗體可包含不存在於受體抗體或供體抗體中的殘基。可進行此等修飾以進一步優化抗體效能。一般而言，人源化抗體將包含至少一個且通常兩個可變域中之實質上所有可變域，其中所有或實質上所有高變環對應於非人類免疫球蛋白之高變環，且所有或實質上所有FR為人類免疫球蛋白序列之FR。人源化抗體可選地亦將包含免疫球蛋白恆定區(Fc)之至少一部分，該恆定區通常為人免疫球蛋白之恆定區。關於更多細節，參見例如Jones等人，Nature 321:522-525 (1986)；Riechmann等人，Nature 332:323-329 (1988)；及Presta, Curr. Op. Struct. Biol. 2:593-596 (1992)。亦參見例如Vaswani及Hamilton，Ann. Allergy, Asthma & Immunol. 1:105-115 (1998)；Harris，Biochem. Soc. Transactions 23:1035-1038 (1995)；Hurle及Gross，Curr. Op. Biotech. 5:428-433 (1994)；及美國專利號6,982,321及7,087,409。"Humanized" forms of non-human (e.g., murine) antibodies are chimeric antibodies containing minimal sequence derived from non-human immunoglobulins. In one embodiment, a humanized antibody is a human immunoglobulin (acceptor antibody) in which residues from the acceptor HVRs are replaced with residues from an HVR of a non-human species (donor antibody) with the desired specificity, affinity, and/or capacity, such as mouse, rat, rabbit, or non-human primate. In some cases, FR residues of the human immunoglobulin are replaced with corresponding non-human residues. In addition, humanized antibodies may include residues that are not present in the acceptor or donor antibodies. Such modifications may be made to further optimize antibody potency. In general, a humanized antibody will comprise substantially all of the variable domains of at least one and typically two variable domains, wherein all or substantially all of the hypervariable loops correspond to those of a non-human immunoglobulin, and all or substantially all of the FRs are FRs of human immunoglobulin sequences. A humanized antibody may also optionally comprise at least a portion of an immunoglobulin constant region (Fc), which is typically that of a human immunoglobulin. For more details, see, e.g., Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992). See also, e.g., Vaswani and Hamilton, Ann. Allergy, Asthma & Immunol. 1:105-115 (1998); Harris, Biochem. Soc. Transactions 23:1035-1038 (1995); Hurle and Gross, Curr. Op. Biotech. 5:428-433 (1994); and U.S. Patent Nos. 6,982,321 and 7,087,409.

「人類抗體」為具有以下胺基酸序列之抗體：對應於由人類所產生及/或已使用製造如本文所揭露之人類抗體之技術中之任一者所製造之抗體的胺基酸序列。該人類抗體的定義具體地排除包含非人類抗原結合殘基的人源化抗體。人類抗體可使用本領域中已知之各種技術(包括噬菌體顯示庫)來產生。Hoogenboom及Winter，J. Mol. Biol. 227:381 (1991)；Marks等人，J. Mol. Biol. 222:581 (1991)。Cole等人，Monoclonal Antibodies and Cancer Therapy，Alan R. Liss，77 (1985)；Boerner等人，J. Immunol. 147(1):86-95 (1991)中所述之方法亦可用於製備人類單株抗體。亦參見van Dijk及van de Winkel，Curr. Opin. Pharmacol. 5: 368-74 (2001)。可藉由將抗原投予轉基因動物(例如，經免疫之異源小鼠)來製備人類抗體，該轉基因動物已被改造以回應於抗原攻擊而產生此類抗體，但其內源性基因座已失去功能(參見例如，美國專利號6,075,181及6,150,584，其等係關於 XENOMOUSETM技術)。亦參見例如，Li等人，Proc. Natl. Acad. Sci. USA 103:3557-3562 (2006)，其係關於經由人類B細胞融合瘤技術生成之人類抗體。A "human antibody" is an antibody having an amino acid sequence that corresponds to an antibody produced by a human and/or that has been produced using any of the techniques for producing human antibodies as disclosed herein. This definition of a human antibody specifically excludes humanized antibodies that contain non-human antigen-binding residues. Human antibodies can be produced using various techniques known in the art, including phage display libraries. Hoogenboom and Winter, J. Mol. Biol. 227:381 (1991); Marks et al., J. Mol. Biol. 222:581 (1991). The methods described in Cole et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, 77 (1985); Boerner et al., J. Immunol. 147(1):86-95 (1991) can also be used to prepare human monoclonal antibodies. See also van Dijk and van de Winkel, Curr. Opin. Pharmacol. 5:368-74 (2001). Human antibodies can be prepared by administering an antigen to a transgenic animal (e.g., an immunized xenogeneic mouse) that has been engineered to produce such antibodies in response to antigenic challenge but in which the endogenous loci have been disabled (see, e.g., U.S. Patent Nos. 6,075,181 and 6,150,584, regarding XENOMOUSE™ technology). See also, e.g., Li et al., Proc. Natl. Acad. Sci. USA 103:3557-3562 (2006) on human antibodies generated by human B-cell hybridoma technology.

當在本文中使用時，術語「高度可變區」、「HVR」或「HV」意指抗體可變域的序列高度變異及/或形成結構上定義的環圈的區域。一般而言，抗體包含六個HVR；三個位於VH中(H1、H2、H3)，且三個位於VL中(L1、L2、L3)。在天然抗體中，H3及L3在六個HVR中表現出最多的多樣性，特別是據信H3在賦予抗體優異特異性方面發揮獨特的作用。參見，例如Xu等人，Immunity 13:37-45 (2000)；Johnson及Wu，在Methods in Molecular Biology 248:1-25 (Lo，編輯，Human Press，Totowa，N.J.，2003)中。實際上，在不存在輕鏈的情況下，僅由重鏈組成的天然駱駝科抗體具有功能及穩定性。參見，例如Hamers-Casterman等人，Nature 363:446-448 (1993)；Sheriff等人，Nature Struct. Biol. 3:733-736 (1996)。HVR亦稱為「CDR」或「互補決定區」。As used herein, the term "hypervariable region," "HVR," or "HV" refers to the regions of the antibody variable domain whose sequences vary greatly and/or form structurally defined loops. Generally, antibodies comprise six HVRs; three are located in the VH (H1, H2, H3) and three are located in the VL (L1, L2, L3). In natural antibodies, H3 and L3 exhibit the greatest diversity among the six HVRs, and H3 in particular is believed to play a unique role in conferring superior specificity to antibodies. See, e.g., Xu et al., Immunity 13:37-45 (2000); Johnson and Wu, in Methods in Molecular Biology 248:1-25 (Lo, ed., Human Press, Totowa, N.J., 2003). In fact, natural Camelidae antibodies composed solely of the heavy chain are functional and stable in the absence of the light chain. See, e.g., Hamers-Casterman et al., Nature 363:446-448 (1993); Sheriff et al., Nature Struct. Biol. 3:733-736 (1996). HVRs are also known as "CDRs" or "complementary determining regions."

免疫球蛋白可變區之結構及位置可參考Kabat, E. A.等人，Sequences of Proteins of Immunological Interest. 第4版，US Department of Health and Human Services. 1987及現可於網際網路(immuno.bme.nwu.edu)上獲得之其更新來確定。The structure and location of immunoglobulin variable regions can be determined by reference to Kabat, E.A. et al., Sequences of Proteins of Immunological Interest. 4th ed., US Department of Health and Human Services, 1987, and updates thereto currently available on the Internet (immuno.bme.nwu.edu).

「框架」或「FR」殘基為如本文定義之HVR殘基以外的彼等可變域殘基。"Framework" or "FR" residues are those variable region residues other than HVR residues as defined herein.

如本文所用，術語「共價連接」意指 經由一或多個化學鍵直接連接或經由一或多個連接子間接連接。任何合適的化學鍵皆可用於產生直接連接，包括但不限於共價鍵(諸如肽鍵及雙硫鍵)或非共價鍵(諸如氫鍵、疏水鍵、離子鍵或凡得瓦鍵)。As used herein, the term "covalently linked" means directly linked via one or more chemical bonds or indirectly linked via one or more linkers. Any suitable chemical bond can be used to create a direct link, including but not limited to covalent bonds (such as peptide bonds and disulfide bonds) or non-covalent bonds (such as hydrogen bonds, hydrophobic bonds, ionic bonds, or van der Waals bonds).

如本文所用，「共價鍵」意指兩個原子之間共用一或多個電子的穩定的鍵。共價鍵之實例包括但不限於肽鍵及雙硫鍵。如本文所用，「肽鍵」意指胺基酸的羧基與相鄰胺基酸的胺基之間形成的共價鍵。如本文所用，「雙硫鍵」意指兩個硫原子之間形成的共價鍵，諸如重鏈片段CH1與輕鏈片段CL藉由一或多個雙硫鍵結合。藉由連接兩個片段中的硫醇基，兩個片段之間可形成一或多個雙硫鍵。在一些實施例中，重鏈片段與輕鏈片段的一或多個半胱胺酸之間可分別形成一或多個雙硫鍵。可藉由兩個硫醇基氧化而形成雙硫鍵。在一些實施例中，該共價連接係藉由共價鍵直接連接。在一些實施例中，該共價連接係藉由肽鍵或雙硫鍵直接連接。As used herein, "covalent bond" means a stable bond between two atoms that share one or more electrons. Examples of covalent bonds include, but are not limited to, peptide bonds and disulfide bonds. As used herein, "peptide bond" means a covalent bond formed between the carboxyl group of an amino acid and the amine group of an adjacent amino acid. As used herein, "disulfide bond" means a covalent bond formed between two sulfur atoms, such as the heavy chain segmentCH1 and the light chain segment CL, which are linked via one or more disulfide bonds. One or more disulfide bonds can be formed between the two segments by linking the thiol groups in the two segments. In some embodiments, one or more disulfide bonds can be formed between one or more cysteine residues in the heavy chain segment and one or more cysteine residues in the light chain segment, respectively. A disulfide bond can be formed by oxidation of two thiol groups. In some embodiments, the covalent linkage is a direct covalent bond. In some embodiments, the covalent linkage is a direct peptide bond or a disulfide bond.

如本文所用，術語「結合」、「特異性地結合至」或「對~具有特異性」意指可測量及可再現之相互作用，諸如標靶與抗體之間的結合，其在分子(包括生物分子)之異質群體存在下由標靶之存在所決定。舉例而言，結合至或特異性地結合至標靶(其可為表位)之抗體為結合此標靶之親和力、親合力、容易性及/或持續時間強於其結合至其他標靶的抗體。在一實施例中，抗體與無關靶標之結合的程度小於抗體與標靶之結合的約10%，例如藉由放射免疫試驗(RIA)的測量。在一些實施例中，特異性地結合至標靶之抗體的解離常數(Kd)為≤ 1 μM、≤100 nM、≤ 10 nM、≤ 1 nM或≤ 0.1 nM。在一些實施例中，抗體特異性地結合至不同物種蛋白質中保守的蛋白質上之表位。在另一實施例中，特異性結合可包括但不要求專一性結合。As used herein, the terms "bind,""specifically bind to," or "specific for" refer to a measurable and reproducible interaction, such as binding between a target and an antibody, that is determined by the presence of the target in the presence of a heterogeneous population of molecules (including biomolecules). For example, an antibody that binds or specifically binds to a target (which may be an epitope) is one that binds to that target with greater affinity, avidity, ease, and/or duration than it binds to other targets. In one embodiment, the extent of binding of the antibody to an unrelated target is less than about 10% of the binding of the antibody to the target, for example, as measured by a radioimmunoassay (RIA). In some embodiments, the dissociation constant (Kd ) of an antibody that specifically binds to a target is ≤ 1 μM, ≤ 100 nM, ≤ 10 nM, ≤ 1 nM, or ≤ 0.1 nM. In some embodiments, the antibody specifically binds to an epitope on a protein that is conserved among proteins of different species. In another embodiment, specific binding may include, but does not require, exclusive binding.

如本文所用，肽、多肽或抗體序列之「胺基酸序列同一性百分比(%)」及「同源性」係定義為：在比對序列及導入缺口(若需要)以獲得最大序列同一性百分比，且不將任何保留性取代視為序列同一性之一部分之後，候選序列中與特定肽或多肽序列中胺基酸殘基一致之胺基酸殘基的百分比。為測定胺基酸序列同一性百分比而進行之比對可以此項技術中熟知之多種方式達成，例如使用公用電腦軟體，諸如BLAST、BLAST-2、ALIGN或MEGALIGNTM(DNASTAR)軟體。具有通常知識者可確定用於量測比對之適當參數，包括為獲得與所比較序列全長範圍內之最大比對所需的任何演算法。As used herein, "percent amino acid sequence identity (%)" and "homology" of peptide, polypeptide, or antibody sequences are defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a particular peptide or polypeptide sequence, after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for determining percent amino acid sequence identity can be achieved in a variety of ways well known in the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN, or MEGALIGN™ (DNASTAR) software. One of ordinary skill in the art can determine appropriate parameters for measuring alignment, including any algorithm necessary to achieve maximum alignment over the full length of the compared sequences.

胺基酸取代可包括但不限於利用一個胺基酸置換多肽中的另一個胺基酸。示例性取代如表B所示。可將胺基酸取代導入感興趣之抗體中，並篩選產物以獲得所需的活性，例如保留/改善抗原結合、降低免疫原性，或改善ADCC或CDC。表B. 示例性胺基酸取代。原始殘基示例性取代Ala (A)Val；Leu；IleArg (R)Lys；Gln；AsnAsn (N)Gln；His；Asp、Lys；ArgAsp (D)Glu；AsnCys (C)Ser；AlaGln (Q)Asn；GluGlu (E)Asp；GlnGly (G)AlaHis (H)Asn；Gln；Lys；ArgIle (I)Leu；Val；Met；Ala；Phe；正白胺酸Leu (L)正白胺酸；Ile；Val；Met；Ala；PheLys (K)Arg；Gln；AsnMet (M)Leu；Phe；IlePhe (F)Trp；Leu；Val；Ile；Ala；TyrPro (P)AlaSer (S)ThrThr (T)Val；SerTrp (W)Tyr；PheTyr (Y)Trp；Phe；Thr；SerVal (V)Ile；Leu；Met；Phe；Ala；正白胺酸Amino acid substitutions may include, but are not limited to, replacing one amino acid with another in a polypeptide. Exemplary substitutions are shown inTableB. Amino acid substitutions can be introduced into antibodies of interest, and the products screened for desired activities, such as retained/improved antigen binding, reduced immunogenicity, or improved ADCC or CDC.TableB. Exemplary amino acid substitutions. Original Residue Exemplary Substitutions Ala (A) Val; Leu; Ile Arg (R) Lys; Gln; Asn Asn (N) Gln; His; Asp, Lys; Arg Asp (D) Glu; Asn Cys (C) Ser; Ala Gln (Q) Asn；Glu Glu (E) Asp; Gln Gly (G) Ala His (H) Asn; Gln; Lys; Arg Ile (I) Leu; Val; Met; Ala; Phe; norleucine Leu (L) norleucine; Ile; Val; Met; Ala; Phe Lys (K) Arg; Gln; Asn Met (M) Leu; Phe; Ile Phe (F) Trp; Leu; Val; Ile; Ala; Tyr Pro (P) Ala Ser (S) Thr Thr (T) Val; Ser Trp (W) Tyr; Phe Tyr (Y) Trp; Phe; Thr; Ser Val (V) Ile; Leu; Met; Phe; Ala; norleucine

可根據常規側鏈性質將胺基酸分組：(1)疏水性：正白胺酸、Met、Ala、Val、Leu、Ile；(2)中性親水性：Cys、Ser、Thr、Asn、Gln；(3)酸性：Asp、Glu；(4)鹼性：His、Lys、Arg；(5)影響鏈位向的殘基：Gly、Pro；及(6)芳香性：Trp、Tyr、Phe。非保留性取代將需要將此等類別之一的成員置換為另一類別的成員。Amino acids can be grouped according to conventional side-chain properties: (1) hydrophobic: norleucine, Met, Ala, Val, Leu, Ile; (2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln; (3) acidic: Asp, Glu; (4) basic: His, Lys, Arg; (5) residues affecting chain orientation: Gly, Pro; and (6) aromatic: Trp, Tyr, Phe. Nonconservative substitutions would require replacing a member of one of these classes with a member of another.

當結合抗體或抗原結合蛋白使用時，術語「多特異性」意指具有多表位特異性的抗體或抗原結合蛋白(亦即，能夠特異性地結合至一個生物分子上的二、三或多個不同表位，或能夠特異性地結合至不同生物分子上的二、三或多個表位)。除非另有說明，否則由多特異性抗體結合之抗原的順序係於多特異性抗體名稱中任意列出。When used in conjunction with an antibody or antigen-binding protein, the term "multispecific" refers to an antibody or antigen-binding protein that has multiple epitope specificities (i.e., it can specifically bind to two, three, or more different epitopes on a single biomolecule, or two, three, or more epitopes on different biomolecules). Unless otherwise indicated, the order of the antigens bound by the multispecific antibody is arbitrarily listed in the name of the multispecific antibody.

當結合抗體或抗原結合蛋白使用時，術語「雙特異性」意指能夠特異性地結合至一個生物分子上的二個不同表位的抗體或抗原結合蛋白，或能夠特異性地結合至兩個不同生物分子上的表位的抗體或抗原結合蛋白。除非另有說明，否則由雙特異性抗體結合之抗原的順序係於雙特異性抗體名稱中任意列出。When used in conjunction with an antibody or antigen-binding protein, the term "bispecific" refers to an antibody or antigen-binding protein that specifically binds to two different epitopes on a single biomolecule, or to epitopes on two different biomolecules. Unless otherwise specified, the order of the antigens bound by the bispecific antibody is arbitrarily listed in the bispecific antibody name.

「杵臼」策略(參見例如PCT國際公開號WO 2006/028936)在說明書的解讀中具有其平常的含義，意指可用於產生全長雙特異性抗體的策略。簡言之，可在影響CH3域相互作用的位置處將形成人類IgG中 CH3域界面的選定胺基酸進行突變以促進異二聚體形成。將具有小型側鏈(臼)的胺基酸導入特異性地結合第一抗原之抗體的重鏈中，且將具有大型側鏈(杵)的胺基酸導入特異性地結合第二抗原之抗體的重鏈中。在共表現兩種抗體之後，由於帶有「臼」的重鏈與帶有「杵」的重鏈優先相互作用而形成異二聚體。The "knob-and-hole" strategy (see, for example, PCT International Publication No. WO 2006/028936) is used in its usual sense in the specification, referring to a strategy that can be used to generate full-length bispecific antibodies. Briefly, selected amino acids that form the CH3 domain interface in human IgG are mutated at positions that influence CH3 domain interactions to promote heterodimer formation. Amino acids with a small side chain (hole) are introduced into the heavy chain of the antibody that specifically binds the first antigen, while amino acids with a large side chain (knob) are introduced into the heavy chain of the antibody that specifically binds the second antigen. Upon co-expression of the two antibodies, heterodimers form due to preferential interaction between the heavy chain with the "hole" and the heavy chain with the "knob."

如本文所用，術語「載體」意指能夠增殖與其連接之另一核酸的核酸分子。該術語包括作為自我複製之核酸結構的載體，以及併入宿主細胞基因體中的載體，其中該載體已被導入。某些載體能夠導引與其可操作地連接之核酸的表現。此類載體在本文中稱為「表現載體」。As used herein, the term "vector" refers to a nucleic acid molecule capable of propagating another nucleic acid to which it is linked. This term includes vectors that are self-replicating nucleic acid structures as well as vectors that are incorporated into the genome of a host cell into which the vector has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operably linked. Such vectors are referred to herein as "expression vectors."

應理解，本文所述之本發明的實施例包括「由……組成」及/或「基本上由……組成」的實施例。It should be understood that embodiments of the present invention described herein include "consisting of" and/or "consisting essentially of" embodiments.

本文中提及的「約」的值或參數包括(及描述)針對該值或參數本身的變異。舉例而言，提及「約X」的描述包括「X」的描述。Reference herein to a value or parameter that is "about" includes (and describes) variations with respect to that value or parameter. For example, a description referring to "about X" includes a description of "X."

術語「約」及「大約」意指在20%之內、15%之內、10%之內、9%之內、8%之內、7%之內、6%之內、5%之內、4%之內、3%之內、2%之內、1%之內或更小的給定值或範圍。The terms "about" and "approximately" mean within 20%, within 15%, within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3%, within 2%, within 1% or less of a given value or range.

如本文所用，提及「非」的值或參數通常意指並描述「除外」的值或參數。舉例而言，該方法不用於治療X型發炎性疾病意指該方法用於治療X型以外的其他類型的發炎性疾病。As used herein, reference to a value or parameter other than "a value or parameter" generally refers to and describes a value or parameter other than "a value or parameter." For example, "the method is not for treating inflammatory diseases of type X" means that the method is for treating inflammatory diseases of other types besides type X.

本文中使用的術語「約X-Y」與「約X至約Y」具有相同的意義。As used herein, the term "about X-Y" has the same meaning as "about X to about Y."

如本文及所附請求項中所用，單數形式「一」、「或」及「該」包括複數參考對象，除非上下文中另有明確規定。II.多特異性抗IL-13抗體構築體As used herein and in the appended claims, the singular forms "a,""an,""or," and "the" include plural references unless the context clearly dictates otherwise.II.Multispecificanti-IL-13antibody constructs

在一些實施例中，提供一種多特異性構築體或分離的抗IL-13抗體構築體，其包含：i)特異性地結合至介白素-13的第一抗體部分(IL-13；「抗IL-13抗體部分」，例如本文所述之抗IL-13抗體部分之任一者)，以及ii)特異性地結合至第二標靶抗原的第二抗體部分。在一些實施例中，該第二標靶抗原為IL-13 (例如，由該第一抗體部分結合的相同表位或不同表位)。在一些實施例中，該第二標靶抗原不為IL-13。在一些實施例中，該第二標靶抗原為TSLP。在一些實施例中，本文所述之該多特異性構築體或分離的抗IL-13抗體構築體包含一或多個(例如，1、2、3、4、5或多個，諸如1或2個)抗IL-13抗體部分(例如，scFv、Fab或全長抗體)。在一些實施例中，該多特異性構築體包含第一及第二抗IL-13抗體部分(例如，第一及第二抗IL-13 Fab或第一及第二抗IL-13 scFv)，可選地特異性地結合至第三標靶抗原的第三抗體部分(例如，Fab、scFv)及可選地特異性地結合至第四標靶抗原的第四抗體部分。在一些實施例中，該多特異性構築體包含：第一抗IL-13抗體部分、特異性地結合至第二標靶抗原的第二抗體部分及特異性地結合至第三標靶抗原的第三抗體部分。在一些實施例中，該第一抗IL-13抗體部分及該第二抗IL-13抗體部分具有相同的胺基酸序列。在一些實施例中，該第一抗IL-13抗體部分及該第二抗IL-13抗體部分 具有不同的胺基酸序列。在一些實施例中，該第一抗IL-13抗體部分及該第二抗IL-13抗體部分結合至相同的IL-13表位。在一些實施例中，該第一抗IL-13抗體部分及該第二抗IL-13抗體部分結合至不同的IL-13表位。在一些實施例中，該第三抗體部分及該第四抗體部分具有相同的胺基酸序列。在一些實施例中，該第三抗體部分及該第四抗體部分具有不同的胺基酸序列。在一些實施例中，該第三抗體部分及該第四抗體部分結合至相同的標靶表位。在一些實施例中，該第三抗體部分及該第四抗體部分結合至不同的標靶表位。In some embodiments, a multispecific construct or isolated anti-IL-13 antibody construct is provided, comprising: i) a first antibody moiety that specifically binds to interleukin-13 (IL-13; "anti-IL-13 antibody moiety," e.g., any of the anti-IL-13 antibody moieties described herein), and ii) a second antibody moiety that specifically binds to a second target antigen. In some embodiments, the second target antigen is IL-13 (e.g., the same epitope or a different epitope bound by the first antibody moiety). In some embodiments, the second target antigen is not IL-13. In some embodiments, the second target antigen is TSLP. In some embodiments, the multispecific constructs or isolated anti-IL-13 antibody constructs described herein comprise one or more (e.g., 1, 2, 3, 4, 5, or more, such as 1 or 2) anti-IL-13 antibody portions (e.g., scFv, Fab, or full-length antibody). In some embodiments, the multispecific construct comprises first and second anti-IL-13 antibody portions (e.g., first and second anti-IL-13 Fabs or first and second anti-IL-13 scFvs), optionally a third antibody portion (e.g., Fab, scFv) that specifically binds to a third target antigen, and optionally a fourth antibody portion that specifically binds to a fourth target antigen. In some embodiments, the multispecific construct comprises a first anti-IL-13 antibody portion, a second antibody portion that specifically binds to a second target antigen, and a third antibody portion that specifically binds to a third target antigen. In some embodiments, the first anti-IL-13 antibody portion and the second anti-IL-13 antibody portion have the same amino acid sequence. In some embodiments, the first anti-IL-13 antibody portion and the second anti-IL-13 antibody portion have different amino acid sequences. In some embodiments, the first anti-IL-13 antibody portion and the second anti-IL-13 antibody portion bind to the same IL-13 epitope. In some embodiments, the first anti-IL-13 antibody portion and the second anti-IL-13 antibody portion bind to different IL-13 epitopes. In some embodiments, the third antibody portion and the fourth antibody portion have the same amino acid sequence. In some embodiments, the third antibody portion and the fourth antibody portion have different amino acid sequences. In some embodiments, the third antibody portion and the fourth antibody portion bind to the same target epitope. In some embodiments, the third antibody portion and the fourth antibody portion bind to different target epitopes.

在一些實施例中，提供一種多特異性構築體或分離的抗IL-13抗體構築體，其包含：i)用於特異性地結合至IL-13的第一抗體工具，以及ii)用於特異性地結合至第二標靶抗原的第二抗體工具。在一些實施例中，提供一種多特異性構築體或分離的抗IL-13抗體構築體，其包含：i)用於特異性地結合至IL-13的第一抗體工具，以及ii)用於特異性地結合至TSLP的第二抗體工具。在一些實施例中，提供一種多特異性構築體或分離的抗IL-13抗體構築體，其包含：i)用於特異性地結合至IL-13的第一抗體工具，以及ii)特異性地結合至第二標靶抗原的第二抗體部分(諸如本文所述之第二抗體部分之任一者)。在一些實施例中，提供一種多特異性構築體或分離的抗IL-13抗體構築體，其包含：i)用於特異性地結合至IL-13的第一抗體工具，以及ii)特異性地結合至TSLP的第二抗體部分(諸如本文所述之抗TSLP部分之任一者)。在一些實施例中，提供一種多特異性構築體或分離的抗IL-13抗體構築體，其包含：i)特異性地結合至IL-13的第一抗體部分(諸如本文所述之抗IL-13抗體部分之任一者)，以及ii)用於特異性地結合至TSLP的第二抗體工具。本文所述之抗IL-13抗體部分可具有一或多個以下有利性質，包括例如：1)對IL-13具有高結合親和力；2)結合至IL-13/IL-13Rα1複合體；3)阻斷IL-4Rα與IL-13Rα1的募集，以防止形成IL-13Rα1/IL-4Rα複合體；及4)經由IL-13Rα1/IL-4Rα複合體抑制IL-13傳訊。本文所述之抗TSLP抗體部分可具有一或多個以下有利性質，包括例如：1)對TSLP具有高結合親和力；2)阻斷TSLP/TSLPR相互作用；及3)經由TSLPR/IL-7Rα複合體抑制TSLP傳訊。In some embodiments, a multispecific construct or isolated anti-IL-13 antibody construct is provided, comprising: i) a first antibody means for specifically binding to IL-13, and ii) a second antibody means for specifically binding to a second target antigen. In some embodiments, a multispecific construct or isolated anti-IL-13 antibody construct is provided, comprising: i) a first antibody means for specifically binding to IL-13, and ii) a second antibody means for specifically binding to TSLP. In some embodiments, a multispecific construct or isolated anti-IL-13 antibody construct is provided, comprising: i) a first antibody moiety for specifically binding to IL-13, and ii) a second antibody moiety that specifically binds to a second target antigen (such as any of the second antibody moieties described herein). In some embodiments, a multispecific construct or isolated anti-IL-13 antibody construct is provided, comprising: i) a first antibody moiety for specifically binding to IL-13, and ii) a second antibody moiety that specifically binds to TSLP (such as any of the anti-TSLP moieties described herein). In some embodiments, a multispecific construct or isolated anti-IL-13 antibody construct is provided, comprising: i) a first antibody moiety (such as any of the anti-IL-13 antibody moieties described herein) that specifically binds to IL-13, and ii) a second antibody moiety for specifically binding to TSLP. The anti-IL-13 antibody moieties described herein may have one or more of the following advantageous properties, including, for example: 1) high binding affinity for IL-13; 2) binding to the IL-13/IL-13Rα1 complex; 3) blocking the recruitment of IL-4Rα and IL-13Rα1 to prevent the formation of the IL-13Rα1/IL-4Rα complex; and 4) inhibiting IL-13 signaling through the IL-13Rα1/IL-4Rα complex. The anti-TSLP antibody portions described herein can have one or more of the following advantageous properties, including, for example: 1) high binding affinity for TSLP; 2) blocking the TSLP/TSLPR interaction; and 3) inhibiting TSLP signaling through the TSLPR/IL-7Rα complex.

在一些實施例中，與單獨的抗IL-13抗體部分(例如，Fab、scFv、全長抗體)相比，本文所述之多特異性構築體具有增加的(例如，增加至少約1.5、2、3、4、5、6、7、8、9、10或更多倍之任一者)體內半衰期。在一些實施例中，與第二抗體部分 特異性地辨識單獨的第二抗原(例如，TSLP)相比，本文所述之多特異性構築體具有增加的(例如，增加至少約1.5、2、3、4、5、6、7、8、9、10或更多倍之任一者)體內半衰期。In some embodiments, the multispecific constructs described herein have increased (e.g., at least about 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more fold) in vivo half-life compared to the anti-IL-13 antibody portion alone (e.g., Fab, scFv, full-length antibody). In some embodiments, the multispecific constructs described herein have increased (e.g., at least about 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more fold) in vivo half-life compared to the second antibody portion that specifically recognizes the second antigen alone (e.g., TSLP).

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13 scFv (例如，本文所述之抗IL-13 scFv之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的全長抗體，其中該抗IL-13 scFv係融合至第二抗體部分之重鏈之一者的C端以形成融合多肽。在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」，例如本文所述之抗IL-13 scFv之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的全長抗體，其中抗IL-13 scFv1係融合至第二抗體部分之第一重鏈的C端以形成第一融合多肽，且抗IL-13 scFv2係融合至第二抗體部分之第二重鏈的C端以形成第二融合多肽。在一些實施例中，該全長抗體之Fc域包含第一次單元及第二次單元。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct is provided, comprising: a first antibody portion that is an anti-IL-13 scFv (e.g., any of the anti-IL-13 scFv described herein), and a second antibody portion that is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP), wherein the anti-IL-13 scFv is fused to the C-terminus of one of the heavy chains of the second antibody portion to form a fusion polypeptide. In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, each of which is a scFv ("anti-IL-13 scFv1" and "anti-IL-13 scFv2," such as any of the anti-IL-13 scFvs described herein), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP), wherein the anti-IL-13 scFv1 is fused to the C-terminus of the first heavy chain of the second antibody portion to form a first fusion polypeptide, and the anti-IL-13 scFv2 is fused to the C-terminus of the second heavy chain of the second antibody portion to form a second fusion polypeptide. In some embodiments, the Fc domain of the full-length antibody comprises a first subunit and a second subunit. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, a subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」，例如本文所述之抗IL-13 scFv之任一者)、兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-抗IL-13 scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-抗IL-13 scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成Fab2。在一些實施例中，該Fc域包含第一次單元及第二次單元。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, which are scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2", e.g., any of the anti-IL-13 scFvs described herein); two second antibody portions, which are Fabs ("Fab1" and "Fab2") that specifically bind to a second target antigen (e.g., TSLP); and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a first polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-anti-IL-13; scFv1-optionally a linker-Fc domain; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optionally a linker-anti-IL-13 scFv2-optionally a linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form Fab2. In some embodiments, the Fc domain comprises the first and second subunits. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的scFv，其中該scFv部分係融合至抗IL-13全長抗體之重鏈之一者的C端以形成融合多肽。在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的scFv (「scFv1」及「scFv2」)，其中scFv1係融合至該抗IL-13全長抗體之第一重鏈的C端以形成第一融合多肽，且scFv2係融合至該抗IL-13全長抗體之第二重鏈的C端以形成第二融合多肽。在一些實施例中，該抗IL-13全長抗體之Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct is provided, comprising: a first antibody portion that is a full-length anti-IL-13 antibody (e.g., any of the full-length anti-IL-13 antibodies described herein), and a second antibody portion that is a scFv that specifically binds to a second target antigen (e.g., TSLP), wherein the scFv portion is fused to the C-terminus of one of the heavy chains of the full-length anti-IL-13 antibody to form a fusion polypeptide. In some embodiments, a multispecific construct is provided, comprising: a first antibody portion, which is a full-length anti-IL-13 antibody (e.g., any of the full-length anti-IL-13 antibodies described herein), and two second antibody portions, which are scFvs ("scFv1" and "scFv2") that specifically bind to a second target antigen (e.g., TSLP), wherein scFv1 is fused to the C-terminus of the first heavy chain of the full-length anti-IL-13 antibody to form a first fusion polypeptide, and scFv2 is fused to the C-terminus of the second heavy chain of the full-length anti-IL-13 antibody to form a second fusion polypeptide. In some embodiments, the Fc domain of the full-length anti-IL-13 antibody is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)、兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的scFv (「scFv1」及「scFv2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；且其中VH1-(H1-CH1)及VL1-(L1-CL)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2。在一些實施例中，該Fc域包含第一次單元及第二次單元。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct is provided that comprises: two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2," such as any of the anti-IL-13 Fabs described herein), two second antibody portions, which are scFvs that specifically bind to a second target antigen (e.g., TSLP), ("scFv1" and "scFv2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optional linker-scFv1-optional linker-Fc domain; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optional linker-scFv2-optional linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); and wherein VH1-(H1-CH1) and VL1-(L1-CL) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-IL-13 Fab2. In some embodiments, the Fc domain comprises a first subunit and a second subunit. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的全長抗體，其中該多特異性構築體包含：i)含有第二抗體部分之第一輕鏈的第一多肽；ii)含有N’至C’：第二抗體部分之第一重鏈-視情況的連接子-VH1-(H1-CH1)的第二多肽；iii)含有N’至C’：第二抗體部分之第二重鏈-視情況的連接子-VH2-(H2-CH1)的第三多肽；iv)含有第二抗體部分之第二輕鏈的第四多肽；v)含有N’至C’：VL1-(L1-CL)的第五多肽；及vi)含有N’至C’：VL2-(L2-CL)的第六多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VL2-(L2-CL)及VH2-(H2-CH1)形成抗IL-13 Fab2。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或(e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該全長抗體包含Fc域，其中該Fc域包含第一次單元及第二次單元。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，該全長抗體之輕鏈係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, each of which is a Fab ("anti-IL-13 Fab1" and "anti-IL-13 Fab2," e.g., any of the anti-IL-13 Fabs described herein), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP), wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain of the second antibody portion; ii) a second polypeptide comprising N' to C': the first heavy chain of the second antibody portion, optionally a linker, VH1-(H1-CH1); iii) a second polypeptide comprising N' to C': the second antibody portion; The invention further comprises the following: iv) a third polypeptide comprising the second heavy chain of the second antibody moiety and, optionally, a linker, VH2-(H2-CH1); iv) a fourth polypeptide comprising the second light chain of the second antibody moiety; v) a fifth polypeptide comprising N' to C': VL1-(L1-CL); and vi) a sixth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 Fab1, and VL2-(L2-CL) and VH2-(H2-CH1) form an anti-IL-13 Fab2. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or (e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the full-length antibody comprises an Fc domain, wherein the Fc domain comprises a first subunit and a second subunit. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, the light chain of the full-length antibody is derived from a kappa light chain.

在一些實施例中，提供一種多特異性構築體，其包含：第一抗IL-13抗體部分，其為全長抗體(「抗IL-13全長抗體」；例如本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」)；其中該多特異性構築體包含：i)含有抗IL-13全長抗體之第一輕鏈(L1)的第一多肽；ii)含有N’至C’：抗IL-13全長抗體之第一重鏈(H1)-視情況的連接子-VH3-(H3-CH1)的第二多肽；iii)含有N’至C’：抗IL-13全長抗體之第二重鏈(H2)-視情況的連接子-VH4-(H4-CH1)的第三多肽；iv)含有抗IL-13全長抗體之第二輕鏈(L2)的第四多肽；v)含有N’至C’：VL3-(L3-CL)的第五多肽；及vi)含有N’至C’：VL4-(L4-CL)的第六多肽；且其中VL3-(L3-CL)及VH3-(H3-CH1)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或(e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗IL-13全長抗體包含Fc域，其中該Fc域包含第一次單元及第二次單元。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，L3-CL及L4-CL係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided that comprises: a first anti-IL-13 antibody portion that is a full-length antibody ("anti-IL-13 full-length antibody"; for example, any of the anti-IL-13 full-length antibodies described herein), and two second antibody portions that are Fabs that specifically bind to a second target antigen (e.g., TSLP). ("Fab1" and "Fab2"); wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain (L1) of an anti-IL-13 full-length antibody; ii) a second polypeptide comprising N' to C': the first heavy chain (H1) of an anti-IL-13 full-length antibody - optionally a linker - VH3 - (H3-CH1); iii) a second polypeptide comprising N' to C': the second heavy chain (H2) of an anti-IL-13 full-length antibody - optionally a linker - VH4 - (H4 -CH1); iv) a fourth polypeptide containing the second light chain (L2) of the anti-IL-13 full-length antibody; v) a fifth polypeptide containing N' to C': VL3-(L3-CL); and vi) a sixth polypeptide containing N' to C': VL4-(L4-CL); wherein VL3-(L3-CL) and VH3-(H3-CH1) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or (e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the full-length anti-IL-13 antibody comprises an Fc domain, wherein the Fc domain comprises a first subunit and a second subunit. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, one subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, L3-CL and L4-CL are derived from a kappa light chain.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的全長抗體；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-第二抗體部分之第一重鏈的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-第二抗體部分之第二重鏈的第三多肽；iv)含有N’至C’：VL2-(L2-CL)的第四多肽；v)含有第二抗體部分之第一輕鏈的第五多肽；及vi)含有第二抗體部分之第二輕鏈的第六多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或(e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該全長抗體包含Fc域，其中該Fc域包含第一次單元及第二次單元。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，該全長抗體之輕鏈係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, each of which is a Fab ("anti-IL-13 Fab1" and "anti-IL-13 Fab2," e.g., any of the anti-IL-13 Fabs described herein), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-a first heavy chain of the second antibody portion; iii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-a first heavy chain of the second antibody portion; ': VH2-(H2-CH1)-optionally a linker-a third polypeptide of the second heavy chain of the second antibody moiety; iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); v) a fifth polypeptide comprising the first light chain of the second antibody moiety; and vi) a sixth polypeptide comprising the second light chain of the second antibody moiety; wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form an anti-IL-13 Fab2. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or (e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the full-length antibody comprises an Fc domain, wherein the Fc domain comprises a first subunit and a second subunit. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, the light chain of the full-length antibody is derived from a kappa light chain.

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」)；其中該多特異性構築體包含：i)含有N’至C’：VL3-(L3-CL)的第一多肽；ii)含有N’至C’：VH3-(H3-CH1)-視情況的連接子-抗IL-13全長抗體之第一重鏈(H1)的第二多肽；iii)含有N’至C’：VH4-(H4-CH1)-視情況的連接子-抗IL-13全長抗體之第二重鏈(H2)的第三多肽；iv)含有N’至C’：VL4-(L4-CL)的第四多肽；v)含有N’至C’：抗IL-13全長抗體之第一輕鏈(L1)的第五多肽；及vi)含有N’至C’：抗IL-13全長抗體之第二輕鏈(L2)的第六多肽；且其中VL3-(L3-CL)及VH3-(H3-CH1)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或(e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該IL-13全長抗體包含Fc域，其中該Fc域包含第一次單元及第二次單元。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，L3-CL及L4-CL係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided, comprising: a first antibody portion that is a full-length IL-13 antibody (e.g., any of the anti-IL-13 full-length antibodies described herein), and two second antibody portions that are Fabs ("Fab1" and "Fab2") that specifically bind to a second target antigen (e.g., TSLP); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL3-(L3-CL); ii) a second polypeptide comprising N' to C': VH3-(H3-CH1)-optionally a linker-a first heavy chain (H1) of an anti-IL-13 full-length antibody; iii) a third polypeptide comprising N' to C': VH4-(H4-CH1)-optionally a linker-a second heavy chain (H2) of an anti-IL-13 full-length antibody; polypeptide; iv) a fourth polypeptide comprising N' to C': VL4-(L4-CL); v) a fifth polypeptide comprising N' to C': the first light chain (L1) of the anti-IL-13 full-length antibody; and vi) a sixth polypeptide comprising N' to C': the second light chain (L2) of the anti-IL-13 full-length antibody; and wherein VL3-(L3-CL) and VH3-(H3-CH1) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or (e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the IL-13 full-length antibody comprises an Fc domain, wherein the Fc domain comprises a first subunit and a second subunit. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, one subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, L3-CL and L4-CL are derived from a kappa light chain.

在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合至第二標靶抗原(例如，TSLP)的第二抗體部分。在一些實施例中，(i)該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)；及/或(ii)該H1包含位於位置126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該H1包含位於位置F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該H1包含F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈(例如，含有SEQ ID NO:74或75之胺基酸序列的λ輕鏈)。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，L2係衍生自κ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈(例如，含有SEQ ID NO:73之胺基酸序列的κ輕鏈)。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，L2係衍生自λ輕鏈。在一些實施例中，該H1包含位於126、170、183、185及220位置處的取代，且該L1包含位於124、135及214位置處的取代，其中該位置係根據EU編號。在一些實施例中，(i)該H1包含位於F126、F170、S183、V185及C220位置處的取代，且該L1包含位於E124、L135及C214位置處的取代；或(ii)該H1包含位於F126、F170、S183、V185及C220位置處的取代，且該L1包含位於Q124、L135及C214位置處的取代；其中該位置係根據EU編號。在一些實施例中，(i)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代；(ii)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代；(iii)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含Q124C、L135F及C214S取代；或(iv)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含Q124C、L135F及C214S取代；且其中該位置係根據EU編號。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，本文所述之H1及L1中的突變可改為在H2及L2上。In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds to a second target antigen (e.g., TSLP). In some embodiments, (i) the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering); and/or (ii) the H1 comprises substitutions at positions 126 and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises an L135F substitution (EU numbering). In some embodiments, the H1 comprises F170V, S183I, and V185L substitutions (EU numbering), and the L1 comprises an L135F substitution (EU numbering). In some embodiments, the L1 is derived from a lambda light chain (e.g., a lambda light chain comprising the amino acid sequence of SEQ ID NO: 74 or 75). In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises F126C and C220S substitutions (EU numbering), and the L1 comprises E124C and C214S substitutions (EU numbering). In some embodiments, L2 is derived from a kappa light chain. In some embodiments, the L1 is derived from a kappa light chain (e.g., a kappa light chain comprising the amino acid sequence of SEQ ID NO: 73). In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions F126C and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C and C214S (EU numbering). In some embodiments, L2 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions 126, 170, 183, 185, and 220, and the L1 comprises substitutions at positions 124, 135, and 214, wherein the positions are according to EU numbering. In some embodiments, (i) the H1 comprises substitutions at positions F126, F170, S183, V185, and C220, and the L1 comprises substitutions at positions E124, L135, and C214; or (ii) the H1 comprises substitutions at positions F126, F170, S183, V185, and C220, and the L1 comprises substitutions at positions Q124, L135, and C214; wherein the positions are numbered according to EU. In some embodiments, (i) the H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and the L1 comprises E124C, L135F, and C214S substitutions; (ii) the H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and the L1 comprises E124C, L135F, and C214S substitutions; (iii) ) the H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and the L1 comprises Q124C, L135F, and C214S substitutions; or (iv) the H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and the L1 comprises Q124C, L135F, and C214S substitutions; and wherein the positions are according to EU numbering. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, one subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, the mutations in H1 and L1 described herein may be altered to H2 and L2.

在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合至第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合至第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合至第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(a)該H1包含F170I、S183L、V185L、T366S、L368A及Y407V取代，該L1包含L135F取代，且該H2包含T366W取代；(b)該H1包含F170I、S183L、V185L及T366W取代，該L1包含L135F取代，且該H2包含T366S、L368A及Y407V取代；(c)該H1包含F170V、S183I、V185L、T366S、L368A及Y407V取代，該L1包含L135F取代，且該H2包含T366W取代；或(d)該H1包含F170V、S183I、V185L及T366W取代，該L1包含L135F取代，且該H2包含T366S、L368A及Y407V取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，L1係衍生自λ輕鏈，且L2係衍生自κ輕鏈。在一些實施例中，L2係衍生自λ輕鏈，且L1係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second heavy chain (H3) comprising VL2 and L1-CL. The invention further comprises a second light chain (L2) of 2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds to a second target antigen (e.g., TSLP); and wherein H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and L1 comprises a substitution at position 135 (EU numbering). In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second subunit comprising VL2 and a second light chain (L2) of L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds to a second target antigen (e.g., TSLP); and wherein the H1 comprises F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises an L135F substitution (EU numbering). In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second subunit comprising VL2 and a second light chain (L2) of L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds to a second target antigen (e.g., TSLP); and wherein H1 comprises F170V, S183I, and V185L substitutions (EU numbering), and L1 comprises an L135F substitution (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, one subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, one subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (a) the H1 comprises F170I, S183L, V185L, T366S, L368A, and Y407V substitutions, the L1 comprises L135F substitutions, and the H2 comprises T366W substitutions; (b) the H1 comprises F170I, S183L, V185L, and T366W substitutions, the L1 comprises L135F substitutions, and the H2 comprises T366S, L368A, and Y407V substitutions; (c) the H (d) wherein H1 comprises F170V, S183I, V185L, T366S, L368A, and Y407V substitutions, L1 comprises an L135F substitution, and H2 comprises a T366W substitution; or (e) wherein H1 comprises F170V, S183I, V185L, and T366W substitutions, L1 comprises an L135F substitution, and H2 comprises T366S, L368A, and Y407V substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, L1 is derived from a lambda light chain, and L2 is derived from a kappa light chain. In some embodiments, L2 is derived from a lambda light chain, and L1 is derived from a kappa light chain.

在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合至第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含位於位置126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(a)該H1包含F126C、C220S、T366S、L368A及Y407V取代，該L1包含E124C及C214S取代，且該H2包含T366W取代；(b)該H1包含F126C、C220S及T366W取代，該L1包含E124C及C214S取代，且該H2包含T366S、L368A及Y407V取代；(c)該H1包含F126C、C220S、T366S、L368A及Y407V取代，該L1包含Q124C及C214S取代，且該H2包含T366W取代；或(d)該H1包含F126C、C220S及T366W取代，該L1包含Q124C及C214S取代，且該H2包含T366S、L368A及Y407V取代；且其中該胺基酸位置係根據EU編號。In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second heavy chain (H3) comprising VL2 and L1-CL. The invention further comprises a second light chain (L2) of 2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds to a second target antigen (e.g., TSLP); and wherein H1 comprises substitutions at positions 126 and 220 (EU numbering), and L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second subunit comprising VL2 and a second light chain (L2) of L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); and wherein H1 comprises F126C and C220S substitutions (EU numbering), and L1 comprises E124C and C214S substitutions (EU numbering). In some embodiments, L2 is derived from a kappa light chain. In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second subunit comprising VL2 and a second light chain (L2) comprising L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); and wherein H1 comprises F126C and C220S substitutions (EU numbering), and L1 comprises Q124C and C214S substitutions (EU numbering). In some embodiments, L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, one subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (a) the H1 comprises F126C, C220S, T366S, L368A, and Y407V substitutions, the L1 comprises E124C and C214S substitutions, and the H2 comprises T366W substitutions; (b) the H1 comprises F126C, C220S, and T366W substitutions, the L1 comprises E124C and C214S substitutions, and the H2 comprises T366S, L368A, and Y407V substitutions; (c) the H 1 comprises F126C, C220S, T366S, L368A and Y407V substitutions, the L1 comprises Q124C and C214S substitutions, and the H2 comprises T366W substitution; or (d) the H1 comprises F126C, C220S and T366W substitutions, the L1 comprises Q124C and C214S substitutions, and the H2 comprises T366S, L368A and Y407V substitutions; and wherein the amino acid positions are according to EU numbering.

在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含位於位置126、170、183、185及220處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，提供一種多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；且其中該H1包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(a)該H1包含F126C、F170I、S183L、V185L、C220S、T366S、L368A及Y407V取代，該L1包含E124C (或Q124C)、L135F及C214S取代，且該H2包含T366W取代；(b)該H1包含F126C、F170I、S183L、V185L、C220S及T366W取代，該L1包含E124C (或Q124C)、L135F及C214S取代，且該H2包含T366S、L368A及Y407V取代；(c)該H1包含F126C、F170V、S183I、V185L、C220S、T366S、L368A及Y407V取代，該L1包含E124C (或Q124C)、L135F及C214S取代，且該H2包含T366W取代；或(d)該H1包含F126C、F170V、S183I、V185L、C220S及T366W取代，該L1包含E124C (或Q124C)、L135F及C214S取代，且該H2包含T366S、L368A及Y407V取代；且其中該胺基酸位置係根據EU編號。In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (H2) comprising VL2 and L2-CL. chain (L2); wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); and wherein H1 comprises substitutions at positions 126, 170, 183, 185, and 220 (EU numbering), and L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain ( L2); wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); and wherein H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions (EU numbering), and L1 comprises E124C, L135F, and C214S substitutions (EU numbering). In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain ( L2); wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); and wherein H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions (EU numbering), and L1 comprises E124C, L135F, and C214S substitutions (EU numbering). In some embodiments, L2 is derived from a kappa light chain. In some embodiments, L1 is derived from a kappa light chain. In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain ( L2); wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); and wherein H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions (EU numbering), and L1 comprises Q124C, L135F, and C214S substitutions (EU numbering). In some embodiments, a multispecific construct is provided, wherein the multispecific construct is a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain ( L2); wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); and wherein H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions (EU numbering), and L1 comprises Q124C, L135F, and C214S substitutions (EU numbering). In some embodiments, L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (a) the H1 comprises F126C, F170I, S183L, V185L, C220S, T366S, L368A, and Y407V substitutions, the L1 comprises E124C (or Q124C), L135F, and C214S substitutions, and the H2 comprises T366W substitutions; (b) the H1 comprises F126C, F170I, S183L, V185L, C220S, and T366W substitutions, the L1 comprises E124C (or Q124C), L135F, and C214S substitutions, and the H2 comprises T366W substitutions. (c) the H1 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A and Y407V substitutions, the L1 comprises E124C (or Q124C), L135F and C214S substitutions, and the H2 comprises T366W substitutions; or (d) the H1 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A and Y407V substitutions, the L1 comprises E124C (or Q124C), L135F and C214S substitutions, and the H2 comprises T366W substitutions, (or Q124C), L135F and C214S substitutions, and the H2 comprises T366S, L368A and Y407V substitutions; and wherein the amino acid positions are according to EU numbering.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合至第二標靶抗原(例如，TSLP)的第二抗體部分；其中該H2包含位於位置170、183及185處的取代(EU編號)，且該L2包含位於位置135處的取代(EU編號)。在一些實施例中，該H2包含位於位置F170、S183及V185處的取代(EU編號)，且該L2包含位於位置L135處的取代(EU編號)。在一些實施例中，該H2包含F170I、S183L及V185L取代(EU編號)，且該L2包含L135F取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL. wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds to a second target antigen (e.g., TSLP); wherein H2 comprises substitutions at positions 170, 183, and 185 (EU numbering), and L2 comprises a substitution at position 135 (EU numbering). In some embodiments, H2 comprises substitutions at positions F170, S183, and V185 (EU numbering), and L2 comprises a substitution at position L135 (EU numbering). In some embodiments, H2 comprises substitutions F170I, S183L, and V185L (EU numbering), and L2 comprises an L135F substitution (EU numbering). In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, a subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該H2包含位於位置170、183及185處的取代(EU編號)，且該L2包含位於位置135處的取代(EU編號)。在一些實施例中，該H2包含位於位置F170、S183及V185處的取代(EU編號)，且該L2包含位於位置L135處的取代(EU編號)。在一些實施例中，該H2包含F170V、S183I及V185L取代(EU編號)，且該L2包含L135F取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL. wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein H2 comprises substitutions at positions 170, 183, and 185 (EU numbering), and L2 comprises a substitution at position 135 (EU numbering). In some embodiments, H2 comprises substitutions at positions F170, S183, and V185 (EU numbering), and L2 comprises a substitution at position L135 (EU numbering). In some embodiments, H2 comprises substitutions F170V, S183I, and V185L (EU numbering), and L2 comprises an L135F substitution (EU numbering). In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, a subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該H2包含位於位置126及220處的取代(EU編號)，且該L2包含位於位置124及214處的取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126及C220處的取代(EU編號)，且該L2包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C及C220S取代(EU編號)，且該L2包含E124C及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126及C220處的取代(EU編號)，且該L2包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C及C220S取代(EU編號)，且該L2包含Q124C及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL. wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein H2 comprises substitutions at positions 126 and 220 (EU numbering), and L2 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, L2 is derived from a lambda light chain. In some embodiments, H2 comprises substitutions at positions F126 and C220 (EU numbering), and L2 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H2 comprises F126C and C220S substitutions (EU numbering), and the L2 comprises E124C and C214S substitutions (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126 and C220 (EU numbering), and the L2 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H2 comprises F126C and C220S substitutions (EU numbering), and the L2 comprises Q124C and C214S substitutions (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該H2包含位於位置126、170、183、185及220處的取代(EU編號)，且該L2包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L2包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L2包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL. ); wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein H2 comprises substitutions at positions 126, 170, 183, 185, and 220 (EU numbering), and L2 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L2 is derived from a lambda light chain. In some embodiments, H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and L2 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H2 comprises F126C, F170I, S183L, V185L, and C220S substitutions (EU numbering), and the L2 comprises E124C, L135F, and C214S substitutions (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L2 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H2 comprises F126C, F170I, S183L, V185L, and C220S substitutions (EU numbering), and the L2 comprises Q124C, L135F, and C214S substitutions (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, a subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該H2包含位於位置126、170、183、185及220處的取代(EU編號)，且該L2包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L2包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L2包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含L234A及L235A取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M428L及N434S取代(EU編號)。在一些實施例中，該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL. ); wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein H2 comprises substitutions at positions 126, 170, 183, 185, and 220 (EU numbering), and L2 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L2 is derived from a lambda light chain. In some embodiments, H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and L2 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H2 comprises F126C, F170V, S183I, V185L, and C220S substitutions (EU numbering), and the L2 comprises E124C, L135F, and C214S substitutions (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L2 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H2 comprises F126C, F170V, S183I, V185L, and C220S substitutions (EU numbering), and the L2 comprises Q124C, L135F, and C214S substitutions (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, a subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering). In some embodiments, a subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」，例如本文所述之抗IL-13 scFv之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的全長抗體；其中抗IL-13 scFv1係融合至第二抗體部分之第一重鏈的C端以形成第一融合多肽，且抗IL-13 scFv2係融合至第二抗體部分之第二重鏈的C端以形成第二融合多肽；且其中該抗IL-13 scFv1及該抗IL-13 scFv2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 scFv1及該抗IL-13 scFv2各自包含：含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。In some embodiments, a multispecific construct is provided, comprising: two anti-IL13 antibody portions, each of which is a scFv ("anti-IL-13 scFv1" and "anti-IL-13 scFv2," e.g., any of the anti-IL-13 scFvs described herein), and a second antibody portion that is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP); wherein the anti-IL-13 scFv1 is fused to the C-terminus of the first heavy chain of the second antibody portion to form a first fusion polypeptide, and the anti-IL-13 scFv2 is fused to the C-terminus of the second heavy chain of the second antibody portion to form a second fusion polypeptide; and wherein the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; In some embodiments, the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise: a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」，例如本文所述之抗IL-13 scFv之任一者)、兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-抗IL-13 scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-抗IL-13 scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；其中VL1-(L1-CL)及VH1-(H1-CH1)形成Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成Fab2；且其中該抗IL-13 scFv1及該抗IL-13 scFv2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 scFv1及該抗IL-13 scFv2各自包含：含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, which are scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2", e.g., any of the anti-IL-13 scFvs described herein); two second antibody portions, which are Fabs ("Fab1" and "Fab2") that specifically bind to a second target antigen (e.g., TSLP); and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a first polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-anti-IL-13; scFv1-optionally a linker-Fc domain; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optionally a linker-anti-IL-13 scFv2-optionally a linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form Fab2; and wherein the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; In some embodiments, the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69.

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的scFv (「scFv1」及「scFv2」)；其中scFv1係融合至該抗IL-13全長抗體之第一重鏈的C端以形成第一融合多肽，且scFv2係融合至該抗IL-13全長抗體之第二重鏈的C端以形成第二融合多肽；且其中該抗IL-13全長抗體包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13全長抗體包含：含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。In some embodiments, a multispecific construct is provided, comprising: a first antibody portion, which is an anti-IL-13 full-length antibody (e.g., any of the anti-IL-13 full-length antibodies described herein), and two second antibody portions, which are scFvs ("scFv1" and "scFv2") that specifically bind to a second target antigen (e.g., TSLP); wherein scFv1 is fused to the C-terminus of the first heavy chain of the anti-IL-13 full-length antibody to form a first fusion polypeptide, and scFv2 is fused to the C-terminus of the second heavy chain of the anti-IL-13 full-length antibody to form a second fusion polypeptide; and wherein the anti-IL-13 full-length antibody comprises: (i) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; In some embodiments, the anti-IL-13 full-length antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-IL-13 full-length antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 125, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 102 or 197. In some embodiments, the anti-IL-13 full-length antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 225, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 102 or 197.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)、兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的scFv (「scFv1」及「scFv2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；其中VH1-(H1-CH1)及VL1-(L1-CL)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2；且其中該抗IL-13 Fab1及該抗IL-13 Fab2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 Fab1及該抗IL-13 Fab2各自包含：含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。In some embodiments, a multispecific construct is provided that comprises: two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2," such as any of the anti-IL-13 Fabs described herein), two second antibody portions, which are scFvs that specifically bind to a second target antigen (e.g., TSLP). ("scFv1" and "scFv2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optional linker-scFv1-optional linker-Fc domain; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optional linker-scFv2-optional linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VH1-(H1-CH1) and VL1-(L1-CL) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-IL-13 Fab2; and wherein the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的全長抗體；其中該多特異性構築體包含：i)含有第二抗體部分之第一輕鏈的第一多肽；ii)含有N’至C’：第二抗體部分之第一重鏈-視情況的連接子-VH1-(H1-CH1)的第二多肽；iii)含有N’至C’：第二抗體部分之第二重鏈-視情況的連接子-VH2-(H2-CH1)的第三多肽；iv)含有第二抗體部分之第二輕鏈的第四多肽；v)含有N’至C’：VL1-(L1-CL)的第五多肽；及vi)含有N’至C’：VL2-(L2-CL)的第六多肽；其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VL2-(L2-CL)及VH2-(H2-CH1)形成抗IL-13 Fab2；且其中該抗IL-13 Fab1及該抗 IL-13 Fab2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 Fab1及該抗IL-13 Fab2各自包含：含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該全長抗體之輕鏈係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2", e.g., any of the anti-IL-13 Fabs described herein), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP); wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain of the second antibody portion; ii) a second polypeptide comprising N' to C': the first heavy chain of the second antibody portion - optionally a linker - VH1-(H1-CH1); iii) a second polypeptide comprising N' to C': the second antibody portion; The invention further comprises a third polypeptide comprising the second heavy chain of the antibody portion and, optionally, a linker, VH2-(H2-CH1); iv) a fourth polypeptide comprising the second light chain of the second antibody portion; v) a fifth polypeptide comprising N' to C': VL1-(L1-CL); and vi) a sixth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 Fab1, and VL2-(L2-CL) and VH2-(H2-CH1) form anti-IL-13 Fab2; and wherein the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the light chain of the full-length antibody is derived from a kappa light chain.

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」)；其中該多特異性構築體包含：i)含有抗IL-13全長抗體之第一輕鏈(L1)的第一多肽；ii)含有N’至C’：抗IL-13全長抗體之第一重鏈(H1)-視情況的連接子-VH3-(H3-CH1)的第二多肽；iii)含有N’至C’：抗IL-13全長抗體之第二重鏈(H2)-視情況的連接子-VH4-(H4-CH1)的第三多肽；iv)含有抗IL-13全長抗體之第二輕鏈(L2)的第四多肽；v)含有N’至C’：VL3-(L3-CL)的第五多肽；及vi)含有N’至C’：VL4-(L4-CL)的第六多肽；其中VL3-(L3-CL)及VH3-(H3-CH1)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2；且其中該抗IL-13全長抗體包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13全長抗體包含：含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或ix)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈。在一些實施例中，L3-CL及L4-CL係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided that comprises: a first antibody portion that is a full-length anti-IL-13 antibody (e.g., any of the full-length anti-IL-13 antibodies described herein), and two second antibody portions that are Fabs that specifically bind to a second target antigen (e.g., TSLP). ("Fab1" and "Fab2"); wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain (L1) of an anti-IL-13 full-length antibody; ii) a second polypeptide comprising N' to C': the first heavy chain (H1) of an anti-IL-13 full-length antibody - optionally a linker - VH3 - (H3-CH1); iii) a third polypeptide comprising N' to C': the second heavy chain (H2) of an anti-IL-13 full-length antibody - optionally a linker - VH4 - (H4-CH1); i v) a fourth polypeptide comprising the second light chain (L2) of the anti-IL-13 full-length antibody; v) a fifth polypeptide comprising N' to C': VL3-(L3-CL); and vi) a sixth polypeptide comprising N' to C': VL4-(L4-CL); wherein VL3-(L3-CL) and VH3-(H3-CH1) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2; and wherein the anti-IL-13 full-length antibody comprises: (i) a polypeptide comprising SEQ In some embodiments, the anti-IL-13 full-length antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 210 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 211 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 212 and two light chains each comprising the amino acid sequence of SEQ ID NO: 213; NO: 212 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vi) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 213 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 225 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; viiii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 101 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; or ix) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 123 and two light chains, each containing an amino acid sequence of SEQ ID NO: 121 or 206. In some embodiments, L3-CL and L4-CL are derived from a kappa light chain.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的全長抗體；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-第二抗體部分之第一重鏈的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-第二抗體部分之第二重鏈的第三多肽；iv)含有N’至C’：VL2-(L2-CL)的第四多肽；v)含有第二抗體部分之第一輕鏈的第五多肽；及vi)含有第二抗體部分之第二輕鏈的第六多肽；其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2；且其中該抗IL-13 Fab1及該抗IL-13 Fab2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 Fab1及該抗IL-13 Fab2各自包含：含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該全長抗體之輕鏈係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2," e.g., any of the anti-IL-13 Fabs described herein), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-a first heavy chain of the second antibody portion; iii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-a first heavy chain of the second antibody portion; C': VH2-(H2-CH1) - optional linker - a third polypeptide of the second heavy chain of the second antibody portion; iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); v) a fifth polypeptide comprising the first light chain of the second antibody portion; and vi) a sixth polypeptide comprising the second light chain of the second antibody portion; wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-IL-13 Fab2; and wherein the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the light chain of the full-length antibody is derived from a kappa light chain.

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」)；其中該多特異性構築體包含：i)含有N’至C’：VL3-(L3-CL)的第一多肽；ii)含有N’至C’：VH3-(H3-CH1)-視情況的連接子-抗IL-13全長抗體之第一重鏈(H1)的第二多肽；iii)含有N’至C’：VH4-(H4-CH1)-視情況的連接子-抗IL-13全長抗體之第二重鏈(H2)的第三多肽；iv)含有N’至C’：VL4-(L4-CL)的第四多肽；v)含有N’至C’：抗IL-13全長抗體之第一輕鏈(L1)的第五多肽；及vi)含有N’至C’：抗IL-13全長抗體之第二輕鏈(L2)的第六多肽；其中VL3-(L3-CL)及VH3-(H3-CH1)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2；且其中該抗IL-13全長抗體包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13全長抗體包含：含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或ix)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈。在一些實施例中，L3-CL及L4-CL係衍生自κ輕鏈。In some embodiments, a multispecific construct is provided that comprises: a first antibody portion that is a full-length anti-IL-13 antibody (e.g., any of the full-length anti-IL-13 antibodies described herein), and two second antibody portions that are Fabs that specifically bind to a second target antigen (e.g., TSLP). ("Fab1" and "Fab2"); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL3-(L3-CL); ii) a second polypeptide comprising N' to C': VH3-(H3-CH1)-optionally a linker-a first heavy chain (H1) of an anti-IL-13 full-length antibody; iii) a third polypeptide comprising N' to C': VH4-(H4-CH1)-optionally a linker-a second heavy chain (H2) of an anti-IL-13 full-length antibody; iv) a third polypeptide comprising N' to C': VH4-(H4-CH1)-optionally a linker-a second heavy chain (H2) of an anti-IL-13 full-length antibody ': VL4-(L4-CL) fourth polypeptide; v) containing N' to C': the first light chain (L1) of the anti-IL-13 full-length antibody; and vi) containing N' to C': the second light chain (L2) of the anti-IL-13 full-length antibody sixth polypeptide; wherein VL3-(L3-CL) and VH3-(H3-CH1) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2; and wherein the anti-IL-13 full-length antibody comprises: (i) containing SEQ In some embodiments, the anti-IL-13 full-length antibody comprises: a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 210 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 211 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 212 and two light chains each comprising the amino acid sequence of SEQ ID NO: 213; NO: 212 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vi) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 213 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 225 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; viiii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 101 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; or ix) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 123 and two light chains, each containing an amino acid sequence of SEQ ID NO: 121 or 206. In some embodiments, L3-CL and L4-CL are derived from a kappa light chain.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該H1包含位於位置F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，該H1含有SEQ ID NO:138或139之胺基酸序列或其與SEQ ID NO:138或139具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L1含有SEQ ID NO:122或207之胺基酸序列或其與SEQ ID NO:122或207具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該H1含有SEQ ID NO:138或139之胺基酸序列，且該L1含有SEQ ID NO:122或207之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the H1 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises an L135F substitution (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 138 or 139, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 138 or 139, and the L1 comprises the amino acid sequence of SEQ ID NO: 122 or 207, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 122 or 207. In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 138 or 139, and the L1 comprises the amino acid sequence of SEQ ID NO: 122 or 207.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該H1包含位於位置F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，該H1含有SEQ ID NO:140或141之胺基酸序列或其與SEQ ID NO:140或141具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L1含有SEQ ID NO:122或207之胺基酸序列或其與SEQ ID NO:122或207具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該H1含有SEQ ID NO:140或141之胺基酸序列，且該L1含有SEQ ID NO:122或207之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the H1 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises F170V, S183I, and V185L substitutions (EU numbering), and the L1 comprises an L135F substitution (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 140 or 141, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 140 or 141, and the L1 comprises the amino acid sequence of SEQ ID NO: 122 or 207, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 122 or 207. In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 140 or 141, and the L1 comprises the amino acid sequence of SEQ ID NO: 122 or 207.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H1包含位於位置126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，該H1含有SEQ ID NO:200或201之胺基酸序列或其與SEQ ID NO:200或201具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L1含有SEQ ID NO:121或206之胺基酸序列或其與SEQ ID NO:121或206具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該H1含有SEQ ID NO:200或201之胺基酸序列，且該L1含有SEQ ID NO:121或206之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H1 comprises substitutions at positions 126 and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions F126C and C220S (EU numbering), and the L1 comprises substitutions at positions E124C and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions F126C and C220S (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 200 or 201, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 200 or 201, and the L1 comprises the amino acid sequence of SEQ ID NO: 121 or 206, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 121 or 206. In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 200 or 201, and the L1 comprises the amino acid sequence of SEQ ID NO: 121 or 206.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H1包含位於位置126、170、183、185及220處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，該H1含有SEQ ID NO:126或127之胺基酸序列或其與SEQ ID NO:126或127具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L1含有SEQ ID NO:120或208之胺基酸序列或其與SEQ ID NO:120或208具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該H1含有SEQ ID NO:126或127之胺基酸序列，且該L1含有SEQ ID NO:120或208之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H1 comprises substitutions at positions 126, 170, 183, 185 and 220 (EU numbering), and the L1 comprises substitutions at positions 124, 135 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 126 or 127, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 126 or 127, and the L1 comprises the amino acid sequence of SEQ ID NO: 120 or 208, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 120 or 208. In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 126 or 127, and the L1 comprises the amino acid sequence of SEQ ID NO: 120 or 208.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H1包含位於位置234、235、428、434、126、170、183、185及220處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，該H1含有SEQ ID NO:128或129之胺基酸序列或其與SEQ ID NO:128或129具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L1含有SEQ ID NO:120或208之胺基酸序列或其與SEQ ID NO:120或208具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該H1含有SEQ ID NO:128或129之胺基酸序列，且該L1含有SEQ ID NO:120或208之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H1 comprises substitutions at positions 234, 235, 428, 434, 126, 170, 183, 185 and 220 (EU numbering), and the L1 comprises substitutions at positions 124, 135 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 128 or 129, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 128 or 129, and the L1 comprises the amino acid sequence of SEQ ID NO: 120 or 208, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 120 or 208. In some embodiments, the H1 comprises the amino acid sequence of SEQ ID NO: 128 or 129, and the L1 comprises the amino acid sequence of SEQ ID NO: 120 or 208.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H2包含位於位置170、183及185處的取代(EU編號)，且該L2包含位於位置135處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該H2包含位於位置F170、S183及V185處的取代(EU編號)，且該L2包含位於位置L135處的取代(EU編號)。在一些實施例中，該H2包含F170I、S183L及V185L取代(EU編號)，且該L2包含L135F取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H2 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L2 comprises a substitution at position 135 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the H2 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L2 comprises a substitution at position L135 (EU numbering). In some embodiments, the H2 comprises F170I, S183L, and V185L substitutions (EU numbering), and the L2 comprises an L135F substitution (EU numbering). In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H2包含位於位置170、183及185處的取代(EU編號)，且該L2包含位於位置135處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該H2包含位於位置F170、S183及V185處的取代(EU編號)，且該L2包含位於位置L135處的取代(EU編號)。在一些實施例中，該H2包含F170V、S183I及V185L取代(EU編號)，且該L2包含L135F取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H2 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L2 comprises a substitution at position 135 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the H2 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L2 comprises a substitution at position L135 (EU numbering). In some embodiments, the H2 comprises F170V, S183I, and V185L substitutions (EU numbering), and the L2 comprises an L135F substitution (EU numbering). In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H2包含位於位置126及220處的取代(EU編號)，且該L2包含位於位置124及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126及C220處的取代(EU編號)，且該L2包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C及C220S取代(EU編號)，且該L2包含E124C及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126及C220處的取代(EU編號)，且該L2包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C及C220S取代(EU編號)，且該L2包含Q124C及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H2 comprises substitutions at positions 126 and 220 (EU numbering), and the L2 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the H2 comprises substitutions at positions F126 and C220 (EU numbering), and the L2 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H2 comprises substitutions at positions F126C and C220S (EU numbering), and the L2 comprises substitutions at positions E124C and C214S (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126 and C220 (EU numbering), and the L2 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H2 comprises substitutions at positions F126C and C220S (EU numbering), and the L2 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H2包含位於位置126、170、183、185及220處的取代(EU編號)，且該L2包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L2包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L2包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H2 comprises substitutions at positions 126, 170, 183, 185 and 220 (EU numbering), and the L2 comprises substitutions at positions 124, 135 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and L2 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, H2 comprises substitutions F126C, F170I, S183L, V185L, and C220S (EU numbering), and L2 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L2 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H2 comprises substitutions F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L2 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合第二標靶抗原(例如，TSLP)的第二抗體部分；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該H2包含位於位置126、170、183、185及220處的取代(EU編號)，且該L2包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L2包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L2包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds a second target antigen (e.g., TSLP); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the H2 comprises substitutions at positions 126, 170, 183, 185 and 220 (EU numbering), and the L2 comprises substitutions at positions 124, 135 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and L2 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, H2 comprises substitutions at positions F126C, F170V, S183I, V185L, and C220S (EU numbering), and L2 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L2 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H2 comprises substitutions F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L2 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, such as human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種多特異性構築體，其包含：兩個第一抗體部分，其等為抗IL-13 scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)，以及第二抗體部分，其為特異性地結合至TSLP的全長抗體(「抗TSLP全長抗體」)，其中抗IL-13 scFv1係經由視情況的連接子融合至抗TSLP全長抗體之第一重鏈的C端以形成第一融合多肽，且抗IL-13 scFv2係經由視情況的連接子融合至抗TSLP全長抗體之第二重鏈的C端以形成第二融合多肽；其中該抗IL-13 scFv1及該抗IL-13 scFv2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該抗TSLP全長抗體包含：(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 scFv1及該抗IL-13 scFv2各自包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗TSLP全長抗體包含：(a)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(c)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(d)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(e)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，該抗IL-13 scFv1及該抗IL-13 scFv2各自含有SEQ ID NO:108之胺基酸序列。在一些實施例中，該抗TSLP全長抗體包含：i)含有SEQ ID NO:104之胺基酸序列的重鏈及含有SEQ ID NO:103之胺基酸序列的輕鏈；或ii)含有SEQ ID NO:105之胺基酸序列的重鏈及含有SEQ ID NO:103之胺基酸序列的輕鏈。在一些實施例中，該連接子含有GG及SEQ ID NO:98-99之任一者的胺基酸序列。在一些實施例中，該抗TSLP全長抗體包含兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈及兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈，且該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:113之胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: two first antibody portions, which are anti-IL-13 scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), and a second antibody portion, which is a full-length antibody that specifically binds to TSLP ("anti-TSLP full-length antibody"), wherein the anti-IL-13 scFv1 is fused to the C-terminus of the first heavy chain of the anti-TSLP full-length antibody via an optional linker to form a first fusion polypeptide, and the anti-IL-13 scFv2 is fused to the C-terminus of the second heavy chain of the anti-TSLP full-length antibody via an optional linker to form a second fusion polypeptide; wherein the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise: (i) a polypeptide comprising SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-TSLP full-length antibody comprises: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. and (vi) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (vii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP full-length antibody comprises: (a) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH comprising the amino acid sequence of SEQ ID NO: 228 and a VL comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise the amino acid sequence of SEQ ID NO: 108. In some embodiments, the full-length anti-TSLP antibody comprises: i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 104 and a light chain comprising the amino acid sequence of SEQ ID NO: 103; or ii) a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 and a light chain comprising the amino acid sequence of SEQ ID NO: 103. In some embodiments, the linker comprises GG and the amino acid sequence of any one of SEQ ID NOs: 98-99. In some embodiments, the full-length anti-TSLP antibody comprises two light chains, each comprising the amino acid sequence of SEQ ID NO: 103 and two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 105, and the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of SEQ ID NO: 113.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)、兩個第二抗體部分，其等為特異性地結合至TSLP的Fab (「抗TSLP Fab1」及「抗TSLP Fab2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-抗IL-13 scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-抗IL-13 scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗TSLP Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗TSLP Fab2；其中該抗IL-13 scFv1及該抗IL-13 scFv2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該抗TSLP Fab1及該抗TSLP Fab2各自包含：(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 scFv1及該抗IL-13 scFv2各自包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗TSLP Fab1及該抗TSLP Fab2各自包含：(a)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(c)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(d)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(e)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，該抗IL-13 scFv1及該抗IL-13 scFv2各自含有SEQ ID NO:108之胺基酸序列。在一些實施例中，該抗TSLP Fab1及該抗TSLP Fab2各自包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。在一些實施例中，該連接子含有獨立地選自GG及SEQ ID NO:98-99之任一者的胺基酸序列。在一些實施例中，該第一多肽及該第四多肽各自含有SEQ ID NO:103之胺基酸序列，且該第二多肽及該第三多肽各自含有SEQ ID NO:110之胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, which are scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), two second antibody portions, which are Fabs that specifically bind to TSLP ("anti-TSLP Fab1" and "anti-TSLP Fab2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a first polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-anti-IL-13; scFv1-optionally a linker-Fc domain; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optionally a linker-anti-IL-13 scFv2-optionally a linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form anti-TSLP Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-TSLP Fab2; wherein the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise: (a) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH comprising the amino acid sequence of SEQ ID NO: 228 and a VL comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the anti-IL-13 scFv1 and the anti-IL-13 scFv2 each comprise the amino acid sequence of SEQ ID NO: 108. In some embodiments, the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103. In some embodiments, the linker comprises an amino acid sequence independently selected from GG and any one of SEQ ID NOs: 98-99. In some embodiments, the first polypeptide and the fourth polypeptide each comprise the amino acid sequence of SEQ ID NO: 103, and the second polypeptide and the third polypeptide each comprise the amino acid sequence of SEQ ID NO: 110.

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13全長抗體，以及兩個第二抗體部分，其等為特異性地結合至TSLP的scFv (「抗TSLP scFv1」及「抗TSLP scFv2」)；其中抗TSLP scFv1係經由視情況的連接子融合至該抗IL-13全長抗體之第一重鏈的C端以形成第一融合多肽，且抗TSLP scFv2係經由視情況的連接子融合至該抗IL-13全長抗體之第二重鏈的C端以形成第二融合多肽；其中該抗IL-13全長抗體包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該抗TSLP scFv1及該抗TSLP scFv2各自包含：(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，抗IL-13全長抗體包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗TSLP scFv1及該抗TSLP scFv2各自包含：(a)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(c)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(d)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(e)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。在一些實施例中，該抗TSLP scFv1及該抗TSLP scFv2各自含有SEQ ID NO:106、107、218、219、226、227及229之任一者的胺基酸序列。在一些實施例中，該連接子含有GG及SEQ ID NO:98-99之任一者的胺基酸序列。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:111之胺基酸序列。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:220-223之任一者的胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: a first antibody portion, which is an anti-IL-13 full-length antibody, and two second antibody portions, which are scFvs that specifically bind to TSLP ("anti-TSLP scFv1" and "anti-TSLP scFv2"); wherein anti-TSLP scFv1 is fused to the C-terminus of the first heavy chain of the anti-IL-13 full-length antibody via an optional linker to form a first fusion polypeptide, and anti-TSLP scFv2 is fused to the C-terminus of the second heavy chain of the anti-IL-13 full-length antibody via an optional linker to form a second fusion polypeptide; wherein the anti-IL-13 full-length antibody comprises: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-TSLP scFv1 and the anti-TSLP scFv2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 full-length antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP scFv1 and the anti-TSLP scFv2 each comprise: (a) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH comprising the amino acid sequence of SEQ ID NO: 228 and a VL comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the full-length anti-IL-13 antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 125, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 102 or 197. In some embodiments, the full-length anti-IL-13 antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 225, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP scFv1 and the anti-TSLP scFv2 each comprise the amino acid sequence of any one of SEQ ID NOs: 106, 107, 218, 219, 226, 227, and 229. In some embodiments, the linker comprises GG and the amino acid sequence of any one of SEQ ID NOs: 98-99. In some embodiments, the full-length anti-IL-13 antibody comprises two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197, wherein the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of SEQ ID NO: 111. In some embodiments, the full-length anti-IL-13 antibody comprises two heavy chains each comprising the amino acid sequence of SEQ ID NO: 225 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197, wherein the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of any one of SEQ ID NOs: 220-223.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13 Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)、兩個第二抗體部分，其等為特異性地結合至TSLP的scFv (「抗TSLP scFv1」及「抗TSLP scFv2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-抗TSLP scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-抗TSLP scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；其中VH1-(H1-CH1)及VL1-(L1-CL)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2；其中該抗IL-13 Fab1及該抗IL-13 Fab2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該抗TSLP scFv1及該抗TSLP scFv2各自包含：(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 Fab1及該抗IL-13 Fab2各自包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗TSLP scFv1及該抗TSLP scFv2各自包含：(a)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(c)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(d)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(e)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，該抗IL-13 Fab1及該抗IL-13 Fab2各自包含含有SEQ ID NO:124之胺基酸序列的第一多肽及含有SEQ ID NO:102或197之胺基酸序列的第二多肽。在一些實施例中，該抗TSLP scFv1及該抗TSLP scFv2各自含有SEQ ID NO:106、107、218、219、226、227及229之任一者的胺基酸序列。在一些實施例中，該連接子含有獨立地選自GG及SEQ ID NO:98-99之任一者的胺基酸序列。在一些實施例中，該第一多肽及該第四多肽各自含有SEQ ID NO:102或197之胺基酸序列，且該第二多肽及該第三多肽各自含有SEQ ID NO:112之胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2"), two second antibody moieties, which are scFvs that specifically bind to TSLP ("anti-TSLP scFv1" and "anti-TSLP scFv2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optional linker-anti-TSLP scFv1-optional linker-Fc domain; iii) a second polypeptide comprising N' to C': VH2-(H2-CH1)-optional linker-anti-TSLP scFv2-optionally a linker-a third polypeptide of the second unit of the Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VH1-(H1-CH1) and VL1-(L1-CL) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form an anti-IL-13 Fab2; wherein the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71 NO:71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:72; and wherein the anti-TSLP scFv1 and the anti-TSLP scFv2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:21; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:22; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:24; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:25; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:26. In some embodiments, the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP scFv1 and the anti-TSLP scFv2 each comprise: (a) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH comprising the amino acid sequence of SEQ ID NO: 228 and a VL comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP scFv1 and the anti-TSLP scFv2 each comprise the amino acid sequence of any one of SEQ ID NOs: 106, 107, 218, 219, 226, 227, and 229. In some embodiments, the linker comprises an amino acid sequence independently selected from GG and any one of SEQ ID NOs: 98-99. In some embodiments, the first polypeptide and the fourth polypeptide each comprise the amino acid sequence of SEQ ID NO: 102 or 197, and the second polypeptide and the third polypeptide each comprise the amino acid sequence of SEQ ID NO: 112.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)，以及第二抗體部分，其為特異性地結合至TSLP的全長抗體(「抗TSLP全長抗體」)；其中該多特異性構築體包含：i)含有抗TSLP全長抗體之第一輕鏈的第一多肽；ii)含有N’至C’：抗TSLP全長抗體之第一重鏈-視情況的連接子-VH1-(H1-CH1)的第二多肽；iii)含有N’至C’：抗TSLP全長抗體之第二重鏈-視情況的連接子-VH2-(H2-CH1)的第三多肽；iv)含有抗TSLP全長抗體之第二輕鏈的第四多肽；v)含有N’至C’：VL1-(L1-CL)的第五多肽；及vi)含有N’至C’：VL2-(L2-CL)的第六多肽；其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VL2-(L2-CL)及VH2-(H2-CH1)形成抗IL-13 Fab2；其中該抗IL-13 Fab1及該抗IL-13 Fab2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該抗TSLP全長抗體包含：(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 Fab1及該抗IL-13 Fab2各自包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗TSLP全長抗體包含：(a)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(c)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(d)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(e)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗TSLP全長抗體之輕鏈係衍生自κ輕鏈。在一些實施例中，該抗TSLP全長抗體包含i)兩條各自含有SEQ ID NO:104之胺基酸序列的重鏈及含有SEQ ID NO:103之胺基酸序列的輕鏈；或ii)含有SEQ ID NO:105之胺基酸序列的重鏈及含有SEQ ID NO:103之胺基酸序列的輕鏈。在一些實施例中，該連接子含有獨立地選自GG及SEQ ID NO:98-99之任一者的胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, each of which is a Fab ("anti-IL-13 Fab1" and "anti-IL-13 Fab2"), and a second antibody portion, which is a full-length antibody that specifically binds to TSLP ("anti-TSLP full-length antibody"); wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain of the anti-TSLP full-length antibody; ii) a second polypeptide comprising N' to C': the first heavy chain of the anti-TSLP full-length antibody - optionally a linker - VH1-(H1-CH1); iii) a second polypeptide comprising N' to C': the anti-TSLP full-length antibody; iv) a fourth polypeptide comprising the second light chain of the anti-TSLP full-length antibody; v) a fifth polypeptide comprising N' to C': VL1-(L1-CL); and vi) a sixth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 antibody. Fab1, and VL2-(L2-CL) and VH2-(H2-CH1) form an anti-IL-13 Fab2; wherein the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-TSLP full-length antibody comprises: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68 NO: 21; (iii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP full-length antibody comprises: (a) a VH comprising the amino acid sequence of SEQ ID NO:63 and a VL comprising the amino acid sequence of SEQ ID NO:53; (b) a VH comprising the amino acid sequence of SEQ ID NO:63 and a VL comprising the amino acid sequence of SEQ ID NO:54; (c) a VH comprising the amino acid sequence of SEQ ID NO:188 and a VL comprising the amino acid sequence of SEQ ID NO:189; (d) a VH comprising the amino acid sequence of SEQ ID NO:188 and a VL comprising the amino acid sequence of SEQ ID NO:190; or (e) a VH comprising the amino acid sequence of SEQ ID NO:228 and a VL comprising the amino acid sequence of SEQ ID NO:190. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the light chain of the anti-TSLP full-length antibody is derived from a kappa light chain. In some embodiments, the full-length anti-TSLP antibody comprises i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 104 and a light chain comprising the amino acid sequence of SEQ ID NO: 103; or ii) a heavy chain comprising the amino acid sequence of SEQ ID NO: 105 and a light chain comprising the amino acid sequence of SEQ ID NO: 103. In some embodiments, the linker comprises an amino acid sequence independently selected from GG and any one of SEQ ID NOs: 98-99.

在一些實施例中，提供一種多特異性構築體，其包含：抗IL-13抗體部分，其為全長抗體(「抗IL-13全長抗體」)，以及兩個第二抗體部分，其等為特異性地結合至TSLP的Fab (「抗TSLP Fab1」及「抗TSLP Fab2」)；其中該多特異性構築體包含：i)含有抗IL-13全長抗體之第一輕鏈(L1)的第一多肽；ii)含有N’至C’：抗IL-13全長抗體之第一重鏈(H1)-視情況的連接子-VH3-(H3-CH1)的第二多肽；iii)含有N’至C’：抗IL-13全長抗體之第二重鏈(H2)-視情況的連接子-VH4-(H4-CH1)的第三多肽；iv)含有抗IL-13全長抗體之第二輕鏈(L2)的第四多肽；v)含有N’至C’：VL3-(L3-CL)的第五多肽；及vi)含有N’至C’：VL4-(L4-CL)的第六多肽；其中VL3-(L3-CL)及VH3-(H3-CH1)形成抗TSLP Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成 抗TSLP Fab2；其中該抗IL-13全長抗體包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該抗TSLP Fab1及該抗TSLP Fab2各自包含：(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13全長抗體包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗TSLP Fab1及該抗TSLP Fab2各自包含：(a)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(c)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(d)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(e)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或ix)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈。在一些實施例中，L3-CL及L4-CL係衍生自κ輕鏈。在一些實施例中，該抗TSLP Fab1及該抗TSLP Fab2各自包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。在一些實施例中，該連接子含有獨立地選自GG及SEQ ID NO:98-99之任一者的胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: an anti-IL-13 antibody portion that is a full-length antibody ("anti-IL-13 full-length antibody"), and two second antibody portions that are Fabs that specifically bind to TSLP ("anti-TSLP Fab1" and "anti-TSLP Fab2"); wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain (L1) of the anti-IL-13 full-length antibody; ii) a second polypeptide comprising N' to C': the first heavy chain (H1) of the anti-IL-13 full-length antibody - optionally a linker - VH3 - (H3-CH1); iii) a second polypeptide comprising N' to C': the second heavy chain (H2) of the anti-IL-13 full-length antibody - optionally a linker - VH3 - (H3-CH1); iv) a fourth polypeptide comprising the second light chain (L2) of the anti-IL-13 full-length antibody; v) a fifth polypeptide comprising N' to C': VL3-(L3-CL); and vi) a sixth polypeptide comprising N' to C': VL4-(L4-CL); wherein VL3-(L3-CL) and VH3-(H3-CH1) form an anti-TSLP Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form an anti-TSLP Fab2; wherein the anti-IL-13 full-length antibody comprises: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. NO: 21; (iii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-IL-13 full-length antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise: (a) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH comprising the amino acid sequence of SEQ ID NO: 228 and a VL comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 210 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 211 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 212 and two light chains each comprising the amino acid sequence of SEQ ID NO: 213; NO: 212 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vi) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 213 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 225 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; viiii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 101 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; or ix) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 123 and two light chains, each containing an amino acid sequence of SEQ ID NO: 121 or 206. In some embodiments, L3-CL and L4-CL are derived from a kappa light chain. In some embodiments, the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103. In some embodiments, the linker comprises an amino acid sequence independently selected from GG and any one of SEQ ID NOs: 98-99.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)，以及第二抗體部分，其為特異性地結合至TSLP的全長抗體(「抗TSLP全長抗體」)；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-抗TSLP全長抗體之第一重鏈的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-抗TSLP全長抗體之第二重鏈的第三多肽；iv)含有N’至C’：VL2-(L2-CL)的第四多肽；v)含有抗TSLP全長抗體之第一輕鏈的第五多肽；及vi)含有抗TSLP全長抗體之第二輕鏈的第六多肽；其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2；其中該抗IL-13 Fab1及該抗IL-13 Fab2各自包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該抗TSLP全長抗體包含：(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13 Fab1及該抗IL-13 Fab2各自包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗TSLP全長抗體包含：(a)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(c)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(d)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(e)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗TSLP全長抗體之輕鏈係衍生自κ輕鏈。在一些實施例中，該抗TSLP全長抗體包含i)兩條各自含有SEQ ID NO:104之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈；或ii)兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈。在一些實施例中，該連接子含有獨立地選自GG及SEQ ID NO:98-99之任一者的胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, each of which is a Fab ("anti-IL-13 Fab1" and "anti-IL-13 Fab2"), and a second antibody portion, which is a full-length antibody that specifically binds to TSLP ("anti-TSLP full-length antibody"); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-a first heavy chain of an anti-TSLP full-length antibody; iii) a second polypeptide comprising N' to C': VH2-(H2-CH1)-optionally a linker-a third polypeptide of the second heavy chain of the anti-TSLP full-length antibody; iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); v) a fifth polypeptide comprising the first light chain of the anti-TSLP full-length antibody; and vi) a sixth polypeptide comprising the second light chain of the anti-TSLP full-length antibody; wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form an anti-IL-13 Fab2; wherein the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-TSLP full-length antibody comprises: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. NO: 21; and (iii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-IL-13 Fab1 and the anti-IL-13 Fab2 each comprise a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP full-length antibody comprises: (a) a VH comprising the amino acid sequence of SEQ ID NO:63 and a VL comprising the amino acid sequence of SEQ ID NO:53; (b) a VH comprising the amino acid sequence of SEQ ID NO:63 and a VL comprising the amino acid sequence of SEQ ID NO:54; (c) a VH comprising the amino acid sequence of SEQ ID NO:188 and a VL comprising the amino acid sequence of SEQ ID NO:189; (d) a VH comprising the amino acid sequence of SEQ ID NO:188 and a VL comprising the amino acid sequence of SEQ ID NO:190; or (e) a VH comprising the amino acid sequence of SEQ ID NO:228 and a VL comprising the amino acid sequence of SEQ ID NO:190. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the light chain of the anti-TSLP full-length antibody is derived from a kappa light chain. In some embodiments, the full-length anti-TSLP antibody comprises i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 104 and two light chains each comprising the amino acid sequence of SEQ ID NO: 103; or ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 105 and two light chains each comprising the amino acid sequence of SEQ ID NO: 103. In some embodiments, the linker comprises an amino acid sequence independently selected from GG and any one of SEQ ID NOs: 98-99.

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13全長抗體，以及兩個第二抗體部分，其等為特異性地結合至TSLP的Fab (「抗TSLP Fab1」及「抗TSLP Fab2」)；其中該多特異性構築體包含：i)含有N’至C’：VL3-(L3-CL)的第一多肽；ii)含有N’至C’：VH3-(H3-CH1)-視情況的連接子-抗IL-13全長抗體之第一重鏈(H1)的第二多肽；iii)含有N’至C’：VH4-(H4-CH1)-視情況的連接子-抗IL-13全長抗體之第二重鏈(H2)的第三多肽；iv)含有N’至C’：VL4-(L4-CL)的第四多肽；v)含有N’至C’：抗IL-13全長抗體之第一輕鏈(L1)的第五多肽；及vi)含有N’至C’：抗IL-13全長抗體之第二輕鏈(L2)的第六多肽；其中VL3-(L3-CL)及VH3-(H3-CH1)形成抗TSLP Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成 抗TSLP Fab2；其中該抗IL-13全長抗體包含：(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:72之胺基酸序列的CDR-L3；且其中該抗TSLP Fab1及該抗TSLP Fab2各自包含：(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；(iv)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(v)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(vi)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13全長抗體包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗TSLP Fab1及該抗TSLP Fab2各自包含：(a)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(c)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(d)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(e)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或ix)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈。在一些實施例中，L3-CL及L4-CL係衍生自κ輕鏈。在一些實施例中，該抗TSLP Fab1及該抗TSLP Fab2各自包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。在一些實施例中，該連接子含有獨立地選自GG及SEQ ID NO:98-99之任一者的胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: a first antibody portion, which is an anti-IL-13 full-length antibody, and two second antibody portions, which are Fabs that specifically bind to TSLP ("anti-TSLP Fab1" and "anti-TSLP Fab2"); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL3-(L3-CL); ii) a second polypeptide comprising N' to C': VH3-(H3-CH1)-optionally a linker-the first heavy chain (H1) of the anti-IL-13 full-length antibody; iii) a second polypeptide comprising N' to C': VH4-(H4-CH1)-optionally a linker-the anti-IL-13 full-length antibody; iv) a fourth polypeptide comprising N' to C': VL4-(L4-CL); v) a fifth polypeptide comprising N' to C': the first light chain (L1) of the anti-IL-13 full-length antibody; and vi) a sixth polypeptide comprising N' to C': the second light chain (L2) of the anti-IL-13 full-length antibody; wherein VL3-(L3-CL) and VH3-(H3-CH1) form an anti-TSLP Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form an anti-TSLP Fab2; wherein the anti-IL-13 full-length antibody comprises: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; and wherein the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise: (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; (iv) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (v) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (vi) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. NO: 21; (iii) CDR-H2 comprising the amino acid sequence of SEQ ID NO: 22; (iv) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (v) CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (vi) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-IL-13 full-length antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise: (a) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH comprising the amino acid sequence of SEQ ID NO: 228 and a VL comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 210 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 211 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 212 and two light chains each comprising the amino acid sequence of SEQ ID NO: 213; NO: 212 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vi) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 213 and two light chains, each containing an amino acid sequence of SEQ ID NO: 120 or 208; vii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 225 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; viiii) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 101 and two light chains, each containing an amino acid sequence of SEQ ID NO: 102 or 197; or ix) two heavy chains, each containing an amino acid sequence of SEQ ID NO: 123 and two light chains, each containing an amino acid sequence of SEQ ID NO: 121 or 206. In some embodiments, L3-CL and L4-CL are derived from a kappa light chain. In some embodiments, the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103. In some embodiments, the linker comprises an amino acid sequence independently selected from GG and any one of SEQ ID NOs: 98-99.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分(例如，本文所述之抗IL-13抗體部分之任一者)，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且其中該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該H1包含位於位置F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:139之胺基酸序列或其與SEQ ID NO:139具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:122或207之胺基酸序列或其與SEQ ID NO:122或207具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:136之胺基酸序列或其與SEQ ID NO:136具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體；或(ii)該H1含有SEQ ID NO:138之胺基酸序列或其與SEQ ID NO:138具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:122或207之胺基酸序列或其與SEQ ID NO:122或207具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:137之胺基酸序列或其與SEQ ID NO:137具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，(i)該H1含有SEQ ID NO:139之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；或(ii)該H1含有SEQ ID NO:138之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion (e.g., any of the anti-IL-13 antibody portions described herein), and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO: 25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and wherein the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO:63 and a VL2 comprising the amino acid sequence of SEQ ID NO:53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO:63 and a VL2 comprising the amino acid sequence of SEQ ID NO:54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO:188 and a VL2 comprising the amino acid sequence of SEQ ID NO:189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO:188 and a VL2 comprising the amino acid sequence of SEQ ID NO:190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO:228 and a VL2 comprising the amino acid sequence of SEQ ID NO:190. In some embodiments, the H1 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises an L135F substitution (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (i) the H1 comprises the amino acid sequence of SEQ ID NO: 139, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 139, the L1 comprises the amino acid sequence of SEQ ID NO: 122 or 207, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 122 or 207, the H2 comprises the amino acid sequence of SEQ ID NO: 136, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 136, and the L2 comprises the amino acid sequence of SEQ ID NO: NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 103; or (ii) the H1 comprises the amino acid sequence of SEQ ID NO: 138, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 138, the L1 comprises the amino acid sequence of SEQ ID NO: 122 or 207, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 122 or 207, and the H2 comprises the amino acid sequence of SEQ ID NO: 137, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: NO:137 has at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity, and L2 comprises the amino acid sequence of SEQ ID NO:103 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO:103. In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 139, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 138, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且其中該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該H1包含位於位置F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:141之胺基酸序列或其與SEQ ID NO:141具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:122或207之胺基酸序列或其與SEQ ID NO:122或207具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:136之胺基酸序列或其與SEQ ID NO:136具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體；或(ii)該H1含有SEQ ID NO:140之胺基酸序列或其與SEQ ID NO:140具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:122或207之胺基酸序列或其與SEQ ID NO:122或207具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:137之胺基酸序列或其與SEQ ID NO:137具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，(i)該H1含有SEQ ID NO:141之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；或(ii)該H1含有SEQ ID NO:140之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO: 25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and wherein the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO:63 and a VL2 comprising the amino acid sequence of SEQ ID NO:53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO:63 and a VL2 comprising the amino acid sequence of SEQ ID NO:54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO:188 and a VL2 comprising the amino acid sequence of SEQ ID NO:189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO:188 and a VL2 comprising the amino acid sequence of SEQ ID NO:190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO:228 and a VL2 comprising the amino acid sequence of SEQ ID NO:190. In some embodiments, the H1 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises F170V, S183I, and V185L substitutions (EU numbering), and the L1 comprises an L135F substitution (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (i) the H1 comprises the amino acid sequence of SEQ ID NO: 141 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 141, the L1 comprises the amino acid sequence of SEQ ID NO: 122 or 207 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 122 or 207, the H2 comprises the amino acid sequence of SEQ ID NO: 136 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 136, and the L2 comprises the amino acid sequence of SEQ ID NO: NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 103; or (ii) the H1 comprises the amino acid sequence of SEQ ID NO: 140, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 140, the L1 comprises the amino acid sequence of SEQ ID NO: 122 or 207, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 122 or 207, and the H2 comprises the amino acid sequence of SEQ ID NO: 137, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: NO:137 has at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity, and L2 comprises the amino acid sequence of SEQ ID NO:103 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO:103. In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 141, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 140, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且其中該H1包含位於位置126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:200之胺基酸序列或其與SEQ ID NO:200具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:121或206之胺基酸序列或其與SEQ ID NO:121或206具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:136之胺基酸序列或其與SEQ ID NO:136具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體；或(ii)該H1含有SEQ ID NO:201之胺基酸序列或其與SEQ ID NO:201具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:121或206之胺基酸序列或其與SEQ ID NO:121或206具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:137之胺基酸序列或其與SEQ ID NO:137具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，(i)該H1含有SEQ ID NO:200之胺基酸序列，該L1含有SEQ ID NO:121或206之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；或(ii)該H1含有SEQ ID NO:201之胺基酸序列，該L1含有SEQ ID NO:121或206之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO: 25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and wherein the H1 comprises substitutions at positions 126 and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO: 228 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions F126C and C220S (EU numbering), and the L1 comprises substitutions at positions E124C and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions F126C and C220S (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (i) the H1 comprises the amino acid sequence of SEQ ID NO: 200, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 200, the L1 comprises the amino acid sequence of SEQ ID NO: 121 or 206, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 121 or 206, the H2 comprises the amino acid sequence of SEQ ID NO: 136, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 136, and the L2 comprises the amino acid sequence of SEQ ID NO: NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 103; or (ii) the H1 comprises the amino acid sequence of SEQ ID NO: 201, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 201, the L1 comprises the amino acid sequence of SEQ ID NO: 121 or 206, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 121 or 206, and the H2 comprises the amino acid sequence of SEQ ID NO: 137, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: NO:137 has at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity, and L2 comprises the amino acid sequence of SEQ ID NO:103 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO:103. In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 200, the L1 contains the amino acid sequence of SEQ ID NO: 121 or 206, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 201, the L1 contains the amino acid sequence of SEQ ID NO: 121 or 206, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且其中該H1包含位於位置126、170、183、185及220處的取代(EU編號)；且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:126之胺基酸序列或其與SEQ ID NO:126具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:120或208之胺基酸序列或其與SEQ ID NO:120或208具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:136之胺基酸序列或其與SEQ ID NO:136具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體；或(ii)該H1含有SEQ ID NO:127之胺基酸序列或其與SEQ ID NO:127具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:120或208之胺基酸序列或其與SEQ ID NO:120或208具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:137之胺基酸序列或其與SEQ ID NO:137具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，(i)該H1含有SEQ ID NO:126之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；或(ii)該H1含有SEQ ID NO:127之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO:25; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:26; and wherein the H1 comprises substitutions at positions 126, 170, 183, 185, and 220 (EU numbering); and the L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO: 228 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (i) the H1 comprises the amino acid sequence of SEQ ID NO: 126, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 126, the L1 comprises the amino acid sequence of SEQ ID NO: 120 or 208, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 120 or 208, the H2 comprises the amino acid sequence of SEQ ID NO: 136, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 136, and the L2 comprises the amino acid sequence of SEQ ID NO: NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 103; or (ii) the H1 comprises the amino acid sequence of SEQ ID NO: 127, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 127, the L1 comprises the amino acid sequence of SEQ ID NO: 120 or 208, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 120 or 208, and the H2 comprises the amino acid sequence of SEQ ID NO: 137, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: NO:137 has at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity, and L2 comprises the amino acid sequence of SEQ ID NO:103 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO:103. In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 126, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 127, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且其中該H1包含位於位置126、170、183、185及220處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:128之胺基酸序列或其與SEQ ID NO:128具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:120或208之胺基酸序列或其與SEQ ID NO:120或208具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:136之胺基酸序列或其與SEQ ID NO:136具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體；或(ii)該H1含有SEQ ID NO:129之胺基酸序列或其與SEQ ID NO:129具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該L1含有SEQ ID NO:120或208之胺基酸序列或其與SEQ ID NO:120或208具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，該H2含有SEQ ID NO:137之胺基酸序列或其與SEQ ID NO:137具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該L2含有SEQ ID NO:103之胺基酸序列或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，(i)該H1含有SEQ ID NO:128之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；或(ii)該H1含有SEQ ID NO:129之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO:25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:26; and wherein the H1 comprises substitutions at positions 126, 170, 183, 185 and 220 (EU numbering), and the L1 comprises substitutions at positions 124, 135 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO: 228 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering). In some embodiments, (i) the H1 comprises the amino acid sequence of SEQ ID NO: 128, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 128, the L1 comprises the amino acid sequence of SEQ ID NO: 120 or 208, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 120 or 208, the H2 comprises the amino acid sequence of SEQ ID NO: 136, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 136, and the L2 comprises the amino acid sequence of SEQ ID NO: NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 103; or (ii) the H1 comprises the amino acid sequence of SEQ ID NO: 129, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 129, the L1 comprises the amino acid sequence of SEQ ID NO: 120 or 208, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 120 or 208, and the H2 comprises the amino acid sequence of SEQ ID NO: 137, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: NO:137 has at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity, and L2 comprises the amino acid sequence of SEQ ID NO:103 or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO:103. In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 128, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 129, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且 其中該H2包含位於位置170、183及185處的取代(EU編號)，且該L2包含位於位置135處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該H2包含位於位置F170、S183及V185處的取代(EU編號)，且該L2包含位於位置L135處的取代(EU編號)。在一些實施例中，該H2包含F170I、S183L及V185L取代(EU編號)，且該L2包含L135F取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽，或其與選自由SEQ ID NO:102或197所組成群組之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO: 25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and wherein the H2 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L2 comprises a substitution at position 135 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO:63 and a VL2 comprising the amino acid sequence of SEQ ID NO:53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO:63 and a VL2 comprising the amino acid sequence of SEQ ID NO:54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO:188 and a VL2 comprising the amino acid sequence of SEQ ID NO:189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO:188 and a VL2 comprising the amino acid sequence of SEQ ID NO:190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO:228 and a VL2 comprising the amino acid sequence of SEQ ID NO:190. In some embodiments, H2 comprises substitutions at positions F170, S183, and V185 (EU numbering), and L2 comprises a substitution at position L135 (EU numbering). In some embodiments, H2 comprises F170I, S183L, and V185L substitutions (EU numbering), and L2 comprises an L135F substitution (EU numbering). In some embodiments, L2 is derived from a lambda light chain. In some embodiments, L2 is derived from a kappa light chain. In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且 其中該H2包含位於位置170、183及185處的取代(EU編號)，且該L2包含位於位置135處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該H2包含位於位置F170、S183及V185處的取代(EU編號)，且該L2包含位於位置L135處的取代(EU編號)。在一些實施例中，該H2包含F170V、S183I及V185L取代(EU編號)，且該L2包含L135F取代(EU編號)。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO: 25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and wherein the H2 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L2 comprises a substitution at position 135 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO:63 and a VL2 comprising the amino acid sequence of SEQ ID NO:53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO:63 and a VL2 comprising the amino acid sequence of SEQ ID NO:54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO:188 and a VL2 comprising the amino acid sequence of SEQ ID NO:189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO:188 and a VL2 comprising the amino acid sequence of SEQ ID NO:190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO:228 and a VL2 comprising the amino acid sequence of SEQ ID NO:190. In some embodiments, H2 comprises substitutions at positions F170, S183, and V185 (EU numbering), and L2 comprises a substitution at position L135 (EU numbering). In some embodiments, H2 comprises F170V, S183I, and V185L substitutions (EU numbering), and L2 comprises an L135F substitution (EU numbering). In some embodiments, L2 is derived from a lambda light chain. In some embodiments, L2 is derived from a kappa light chain. In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且 其中該H2包含位於位置126及220處的取代(EU編號)，且該L2包含位於位置124及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126及C220處的取代(EU編號)，且該L2包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C及C220S取代(EU編號)，且該L2包含E124C及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126及C220處的取代(EU編號)，且該L2包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C及C220S取代(EU編號)，且該L2包含Q124C及C214S取代(EU編號)。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO: 25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 26; and wherein the H2 comprises substitutions at positions 126 and 220 (EU numbering), and the L2 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO: 228 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the H2 comprises substitutions at positions F126 and C220 (EU numbering), and the L2 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H2 comprises substitutions at positions F126C and C220S (EU numbering), and the L2 comprises substitutions at positions E124C and C214S (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126 and C220 (EU numbering), and the L2 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H2 comprises substitutions at positions F126C and C220S (EU numbering), and the L2 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且 其中該H2包含位於位置126、170、183、185及220處的取代(EU編號)；且該L2包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L2包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L2包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO:25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:26; and wherein the H2 comprises substitutions at positions 126, 170, 183, 185 and 220 (EU numbering); and the L2 comprises substitutions at positions 124, 135 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO: 228 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and L2 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, H2 comprises substitutions at positions F126C, F170I, S183L, V185L, and C220S (EU numbering), and L2 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L2 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H2 comprises substitutions F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L2 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且其中該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3；其中該抗TSLP抗體部分包含：VH2，其包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3，以及VL2，其包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3；且 其中該H2包含位於位置126、170、183、185及220處的取代(EU編號)，且該L2包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:53之胺基酸序列的VL2；(b)含有SEQ ID NO:63之胺基酸序列的VH2及含有SEQ ID NO:54之胺基酸序列的VL2；(c)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:189之胺基酸序列的VL2；(d)含有SEQ ID NO:188之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2；或(e)含有SEQ ID NO:228之胺基酸序列的VH2及含有SEQ ID NO:190之胺基酸序列的VL2。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L2包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該H2包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L2包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H2包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L2包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4，例如人類IgG1。在一些實施例中，該Fc域之每一次單元包含：(i) L234A及L235A取代(EU編號)；(ii) M428L及N434S取代(EU編號)；及/或(iii) M252Y、S254T及T256E取代(EU編號)。在一些實施例中，(i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或(ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; NO: 67; (iii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 68; and wherein the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72; wherein the anti-TSLP antibody portion comprises: VH2 comprising (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22, and VL2 comprising (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 NO:25; and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:26; and wherein the H2 comprises substitutions at positions 126, 170, 183, 185 and 220 (EU numbering), and the L2 comprises substitutions at positions 124, 135 and 214 (EU numbering). In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 69. In some embodiments, the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 53; (b) a VH2 comprising the amino acid sequence of SEQ ID NO: 63 and a VL2 comprising the amino acid sequence of SEQ ID NO: 54; (c) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 189; (d) a VH2 comprising the amino acid sequence of SEQ ID NO: 188 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190; or (e) a VH2 comprising the amino acid sequence of SEQ ID NO: 228 and a VL2 comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and L2 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, H2 comprises substitutions at positions F126C, F170V, S183I, V185L, and C220S (EU numbering), and L2 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, L2 is derived from a kappa light chain. In some embodiments, the H2 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L2 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H2 comprises substitutions F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L2 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the anti-IL-13 antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 102 or 197. In some embodiments, the anti-TSLP antibody portion comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 109, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 103. In some embodiments, the Fc domain is derived from an IgG selected from the group consisting of IgG1, IgG2, IgG3, and IgG4, e.g., human IgG1. In some embodiments, each subunit of the Fc domain comprises: (i) L234A and L235A substitutions (EU numbering); (ii) M428L and N434S substitutions (EU numbering); and/or (iii) M252Y, S254T, and T256E substitutions (EU numbering). In some embodiments, (i) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or (ii) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)，以及第二抗體部分，其為特異性地結合至TSLP的全長抗體(「抗TSLP全長抗體」)；其中抗IL-13 scFv1係經由連接子融合至抗TSLP全長抗體之第一重鏈的C端以形成第一融合多肽，且抗IL-13 scFv2係經由連接子融合至抗TSLP全長抗體之第二重鏈的C端以形成第二融合多肽；其中該抗TSLP全長抗體包含兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈及兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:113之胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, each of which is a scFv ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), and a second antibody portion, which is a full-length antibody that specifically binds to TSLP ("anti-TSLP full-length antibody"); wherein the anti-IL-13 scFv1 is fused to the C-terminus of the first heavy chain of the anti-TSLP full-length antibody via a linker to form a first fusion polypeptide, and the anti-IL-13 scFv2 is fused to the C-terminus of the second heavy chain of the anti-TSLP full-length antibody via a linker to form a second fusion polypeptide; wherein the anti-TSLP full-length antibody comprises two light chains, each comprising the amino acid sequence of SEQ ID NO: 103, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 104. The invention relates to a recombinant protein comprising the amino acid sequence of SEQ ID NO: 105, wherein the first fusion polypeptide and the second fusion polypeptide each contain the amino acid sequence of SEQ ID NO: 113.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)、兩個第二抗體部分，其等為特異性地結合至TSLP的Fab (「抗TSLP Fab1」及「抗TSLP Fab2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-連接子-抗IL-13 scFv1-連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-連接子-抗IL-13 scFv2-連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗TSLP Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗TSLP Fab2；且其中該第一多肽及該第四多肽各自含有SEQ ID NO:103之胺基酸序列，且該第二多肽及該第三多肽各自含有SEQ ID NO:110之胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, which are scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), two second antibody portions, which are Fabs that specifically bind to TSLP ("anti-TSLP Fab1" and "anti-TSLP Fab2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising a first subunit comprising N' to C': VH1-(H1-CH1)-linker-anti-IL-13 scFv1-linker-Fc domain; iii) a second polypeptide comprising N' to C': VH2-(H2-CH1)-linker-anti-IL-13 iv) a third polypeptide of the second subunit of the scFv2-linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form anti-TSLP Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-TSLP Fab2; and wherein the first polypeptide and the fourth polypeptide each contain the amino acid sequence of SEQ ID NO: 103, and the second polypeptide and the third polypeptide each contain the amino acid sequence of SEQ ID NO: 110.

在一些實施例中，提供一種多特異性構築體，其包含：第一抗體部分，其為抗IL-13全長抗體，以及兩個第二抗體部分，其等為特異性地結合至TSLP的scFv (「抗TSLP scFv1」及「抗TSLP scFv2」)；其中抗TSLP scFv1係經由連接子融合至抗IL-13全長抗體之第一重鏈的C端以形成第一融合多肽，且抗TSLP scFv2係經由連接子融合至抗IL-13全長抗體之第二重鏈的C端以形成第二融合多肽；(i)其中該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:111之胺基酸序列；或(ii)其中該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:220-223之任一者的胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: a first antibody portion, which is an anti-IL-13 full-length antibody, and two second antibody portions, which are scFvs that specifically bind to TSLP ("anti-TSLP scFv1" and "anti-TSLP scFv2"); wherein anti-TSLP scFv1 is fused to the C-terminus of the first heavy chain of the anti-IL-13 full-length antibody via a linker to form a first fusion polypeptide, and anti-TSLP scFv2 is fused to the C-terminus of the second heavy chain of the anti-IL-13 full-length antibody via a linker to form a second fusion polypeptide; (i) wherein the anti-IL-13 full-length antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 125, and two second chains, each comprising the amino acid sequence of SEQ ID NO: 126; NO: 102 or 197, and wherein the first fusion polypeptide and the second fusion polypeptide each contain the amino acid sequence of SEQ ID NO: 111; or (ii) wherein the anti-IL-13 full-length antibody comprises two heavy chains, each containing the amino acid sequence of SEQ ID NO: 225 and two light chains, each containing the amino acid sequence of SEQ ID NO: 102 or 197, and wherein the first fusion polypeptide and the second fusion polypeptide each contain the amino acid sequence of any one of SEQ ID NOs: 220-223.

在一些實施例中，提供一種多特異性構築體，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)、兩個第二抗體部分，其等為特異性地結合至TSLP的scFv (「抗TSLP scFv1」及「抗TSLP scFv2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-連接子-抗TSLP scFv1-連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-連接子-抗TSLP scFv2-連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；其中VH1-(H1-CH1)及VL1-(L1-CL)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2；且其中該第一多肽及該第四多肽各自含有SEQ ID NO:102或197之胺基酸序列，且該第二多肽及該第三多肽各自含有SEQ ID NO:112之胺基酸序列。In some embodiments, a multispecific construct is provided, comprising: two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2"), two second antibody portions, which are scFvs that specifically bind to TSLP ("anti-TSLP scFv1" and "anti-TSLP scFv2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising a first subunit comprising N' to C': VH1-(H1-CH1)-linker-anti-TSLP scFv1-linker-Fc domain; iii) a second polypeptide comprising N' to C': VH2-(H2-CH1)-linker-anti-TSLP iv) a third polypeptide of the second subunit of the scFv2-linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VH1-(H1-CH1) and VL1-(L1-CL) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form an anti-IL-13 Fab2; and wherein the first polypeptide and the fourth polypeptide each comprise the amino acid sequence of SEQ ID NO: 102 or 197, and the second polypeptide and the third polypeptide each comprise the amino acid sequence of SEQ ID NO: 112.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該H1包含F170I、S183L、V185L、L234A、L235A、M428L及N434S取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A、Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A、Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:139之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列，或(ii)該H1含有SEQ ID NO:138之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second heavy chain (H3) comprising VL2 and L2-CH1. The invention further comprises a second light chain (L2) of CL; wherein H1 and L1 form an anti-IL-13 antibody, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein H1 comprises F170I, S183L, V185L, L234A, L235A, M428L, and N434S substitutions (EU numbering), and L1 comprises an L135F substitution (EU numbering). In some embodiments, (i) H1 further comprises a T366W substitution (EU numbering), and H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and H2 comprises a T366W substitution (EU numbering). In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 139, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103, or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 138, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該H1包含F170V、S183I、L234A、L235A、M428L N434S及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A、Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A、Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:141之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列，或(ii)該H1含有SEQ ID NO:140之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein H1 comprises F170V, S183I, L234A, L235A, M428L, N434S and V185L substitutions (EU numbering), and the L1 comprises an L135F substitution (EU numbering). In some embodiments, (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 141, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103, or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 140, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該H1包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、M428L及N434S取代(EU編號)；且該L1包含E124C或Q124C、L135F及C214S取代(EU編號)。在一些實施例中，(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A、Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A、Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:126之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列，或(ii)該H1含有SEQ ID NO:127之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein H1 comprises F126C, F170I, S183L, V185L, C220S, L234A, L235A, M428L, and N434S substitutions (EU numbering); and L1 comprises E124C or Q124C, L135F, and C214S substitutions (EU numbering). In some embodiments, (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 126, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103, or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 127, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，提供一種含有異二聚體雙特異性抗體的多特異性構築體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至TSLP的第二抗體部分(「抗TSLP抗體部分」)；其中該H1包含F126C、F170V、S183I、C220S、L234A、L235A、M428L N434S及V185L取代(EU編號)，且該L1包含E124C或Q124C、L135F及C214S取代(EU編號)。在一些實施例中，(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A、Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A、Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，(i)該H1含有SEQ ID NO:128之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列，或(ii)該H1含有SEQ ID NO:129之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。In some embodiments, a multispecific construct comprising a heterodimeric bispecific antibody is provided, comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain; iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form an anti-IL-13 antibody portion, and H2 and L2 form a second antibody portion that specifically binds to TSLP (the "anti-TSLP antibody portion"); wherein H1 comprises F126C, F170V, S183I, C220S, L234A, L235A, M428L N434S and V185L substitutions (EU numbering), and the L1 comprises E124C or Q124C, L135F, and C214S substitutions (EU numbering). In some embodiments, (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, (i) the H1 contains the amino acid sequence of SEQ ID NO: 128, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103, or (ii) the H1 contains the amino acid sequence of SEQ ID NO: 129, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

本發明進一步提供融合蛋白，其包含本文所述之多特異性構築體之任一者、多特異性構築體接合物(例如，小分子藥物接合物)或表現本文所述之多特異性構築體之任一者的分離細胞。在一些實施例中，本文所述之多特異性構築體之任一者之一或多個多肽的N端及/或C端可含有用於蛋白質純化的組胺酸標籤(HIS-標籤)。抗IL-13抗體部分The present invention further provides fusion proteins comprising any of the multispecific constructs described herein, multispecific construct conjugates (e.g., small molecule drug conjugates), or isolated cells expressing any of the multispecific constructs described herein. In some embodiments, the N-terminus and/or C-terminus of one or more polypeptides of any of the multispecific constructs described herein may contain a histidine tag (HIS-tag) for protein purification.Anti-IL-13AntibodyPortion

IL-13為由不同T細胞子群及樹突細胞產生的細胞激素。IL-13涉及Th2發炎反應，並已被確定為治療氣喘的可能治療標靶。IL-13可用作特定發炎性疾病(例如，過敏或自體免疫疾病)發展的生物標記、預後指標及抗原表現發炎性疾病的免疫治療標靶。IL-13 is a cytokine produced by various T cell subsets and dendritic cells. IL-13 is involved in the Th2 inflammatory response and has been identified as a potential therapeutic target for asthma. IL-13 can be used as a biomarker for the development of specific inflammatory diseases (e.g., allergies or autoimmune diseases), a prognostic indicator, and as a target for immunotherapy of antigen-expressing inflammatory diseases.

本文所述之抗IL-13抗體部分之任一者可用作本文所述之分離的抗IL-13抗體構築體(例如，多特異性構築體)之任一者的第一、第二、第三、第四或更多抗體部分。Any of the anti-IL-13 antibody portions described herein can be used as the first, second, third, fourth, or more antibody portions of any of the isolated anti-IL-13 antibody constructs (e.g., multispecific constructs) described herein.

在一些實施例中，提供一種含有本文所述之抗IL-13抗體部分之任一者的多特異性構築體(例如，單特異性、多特異性、單價或多價) 。在一些實施例中，該多特異性構築體基本上由以下組成(或由以下組成)：抗IL-13抗體部分(例如，抗IL-13全長抗體、抗IL-13 Fab、抗IL-13 scFv)，例如本文所述之抗IL-13抗體部分之任一者。因此，在一些實施例中亦提供分離的抗IL-13抗體部分，諸如本文所述之抗IL-13抗體部分之任一者。In some embodiments, a multispecific construct (e.g., monospecific, multispecific, monovalent, or multivalent) comprising any of the anti-IL-13 antibody portions described herein is provided. In some embodiments, the multispecific construct consists essentially of (or consists of) an anti-IL-13 antibody portion (e.g., an anti-IL-13 full-length antibody, an anti-IL-13 Fab, an anti-IL-13 scFv), such as any of the anti-IL-13 antibody portions described herein. Thus, in some embodiments, an isolated anti-IL-13 antibody portion, such as any of the anti-IL-13 antibody portions described herein, is also provided.

在一些實施例中，該分離的抗IL-13抗體構築體係單特異性。在一些實施例中，該分離的抗IL-13抗體構築體為多特異性(例如，雙特異性)，在本文中可與「多特異性抗IL-13抗體構築體」、「多特異性抗體構築體」或「多特異性構築體」互換使用。在一些實施例中，該分離的抗IL-13抗體構築體為單價。在一些實施例中，該分離的抗IL-13抗體構築體為多價。在一些實施例中，該分離的抗IL-13抗體構築體係多價或單特異性。在一些實施例中，該分離的抗IL-13抗體構築體係多價及多特異性。In some embodiments, the isolated anti-IL-13 antibody construct is monospecific. In some embodiments, the isolated anti-IL-13 antibody construct is multispecific (e.g., bispecific), which are used interchangeably herein with "multispecific anti-IL-13 antibody construct," "multispecific antibody construct," or "multispecific construct." In some embodiments, the isolated anti-IL-13 antibody construct is monovalent. In some embodiments, the isolated anti-IL-13 antibody construct is multivalent. In some embodiments, the isolated anti-IL-13 antibody construct is multivalent or monospecific. In some embodiments, the isolated anti-IL-13 antibody construct is multivalent and multispecific.

在一些實施例中，該分離的抗IL-13抗體構築體係選自由以下組成之群組：多特異性抗體構築體(例如，雙特異性抗體構築體)、抗體檢測標籤接合物、免疫細胞激素及串聯雙價抗體。In some embodiments, the isolated anti-IL-13 antibody construct is selected from the group consisting of a multispecific antibody construct (e.g., a bispecific antibody construct), an antibody detection tag conjugate, an immunocytokine, and a tandem bivalent antibody.

本發明進一步提供融合蛋白，其包含本文所述之抗IL-13抗體構築體(例如，多特異性構築體)或抗IL-13抗體部分之任一者以及本文所述之多特異性構築體、抗IL-13抗體構築體或抗IL-13抗體部分之任一者的接合物(例如，檢測標籤接合物)。The present invention further provides fusion proteins comprising any of the anti-IL-13 antibody constructs (e.g., multispecific constructs) or anti-IL-13 antibody portions described herein and a conjugate (e.g., a detection tag conjugate) of any of the multispecific constructs, anti-IL-13 antibody constructs, or anti-IL-13 antibody portions described herein.

在一些實施例中，該抗IL-13抗體部分特異性地結合至IL-13分子的標靶表位。IL-13抗原可來自各種動物物種，包括人類、非人類靈長類、小鼠、大鼠、兔或其他非人類哺乳動物。在一些實施例中，該IL-13分子為人類IL-13。在一些實施例中，該IL-13分子為小鼠IL-13。在一些實施例中，該IL-13分子為食蟹獼猴IL-13。在一些實施例中，該IL-13分子為野生型。在一些實施例中，該IL-13分子為突變體(例如，與來自同一物種的野生型IL-13序列相比，包含一或多個插入、缺失及/或胺基酸取代)。在一些實施例中，該IL-13分子包含R110Q 突變。R110Q為天然存在的IL-13多型，其與總血清IgE含量增加及氣喘密切相關。在一些實施例中，該抗IL-13抗體部分與人類IL-13及食蟹獼猴IL-13交叉反應。在一些實施例中，該抗IL-13抗體部分結合至野生型人類IL-13及含有R110Q突變的人類IL-13。In some embodiments, the anti-IL-13 antibody portion specifically binds to a target epitope of an IL-13 molecule. IL-13 antigens can be derived from various animal species, including humans, non-human primates, mice, rats, rabbits, or other non-human mammals. In some embodiments, the IL-13 molecule is human IL-13. In some embodiments, the IL-13 molecule is mouse IL-13. In some embodiments, the IL-13 molecule is cynomolgus macaque IL-13. In some embodiments, the IL-13 molecule is wild-type. In some embodiments, the IL-13 molecule is a mutant (e.g., comprising one or more insertions, deletions, and/or amino acid substitutions compared to the wild-type IL-13 sequence from the same species). In some embodiments, the IL-13 molecule comprises the R110Q mutation. R110Q is a naturally occurring polymorph of IL-13 that is closely associated with increased total serum IgE levels and asthma. In some embodiments, the anti-IL-13 antibody portion cross-reacts with human IL-13 and cynomolgus macaque IL-13. In some embodiments, the anti-IL-13 antibody portion binds to wild-type human IL-13 and human IL-13 containing the R110Q mutation.

在一些實施例中，該抗IL-13抗體部分防止IL-13與IL-13Rα1相互作用。在一些實施例中，該抗IL-13抗體部分防止IL-13與IL-13Rα2 (IL-13誘餌受體)相互作用。在一些實施例中，該抗IL-13抗體部分防止IL-13與IL-13Rα1及IL-13Rα2 (阻斷IL-13傳訊及內生性IL-13調節)相互作用，本文中亦稱為「第I類抗IL-13抗體」。在一些實施例中，該抗IL-13抗體部分阻斷與IL-4Rα形成IL-13/IL-13Rα1複合體(因此阻斷傳訊)或中和IL-13Rα1及IL-4Rα上的IL-13活性，本文中亦稱為「第II類抗IL-13抗體」。在一些實施例中，本文所述之抗IL-13抗體部分(例如，73P1)為第II類抗IL-13抗體。In some embodiments, the anti-IL-13 antibody portion prevents IL-13 from interacting with IL-13Rα1. In some embodiments, the anti-IL-13 antibody portion prevents IL-13 from interacting with IL-13Rα2 (the IL-13 decoy receptor). In some embodiments, the anti-IL-13 antibody portion prevents IL-13 from interacting with both IL-13Rα1 and IL-13Rα2 (blocking IL-13 signaling and endogenous IL-13 regulation), also referred to herein as a "Class I anti-IL-13 antibody." In some embodiments, the anti-IL-13 antibody portion blocks the formation of an IL-13/IL-13Rα1 complex with IL-4Rα (thereby blocking signaling) or neutralizes IL-13 activity on IL-13Rα1 and IL-4Rα, and is also referred to herein as a "Class II anti-IL-13 antibody." In some embodiments, the anti-IL-13 antibody portion described herein (e.g., 73P1) is a Class II anti-IL-13 antibody.

本文所述之多特異性構築體包含一或多個(例如，1、2、3、4、5或多個，諸如1或2個)特異性地結合至IL-13的抗IL-13抗體部分(例如，全長抗體、scFv、Fab)。在一些實施例中，該多特異性構築體包含二或多個抗IL-13抗體部分(例如，scFv，例如Fab)。The multispecific constructs described herein comprise one or more (e.g., 1, 2, 3, 4, 5, or more, such as 1 or 2) anti-IL-13 antibody portions (e.g., full-length antibody, scFv, Fab) that specifically bind to IL-13. In some embodiments, the multispecific constructs comprise two or more anti-IL-13 antibody portions (e.g., scFv, such as Fab).

在一些實施例中，與單獨的抗IL-13抗體部分(例如，Fab、scFv)相比，本文所述之多特異性構築體具有增加的(例如，增加至少約1.5、2、3、4、5、6、7、8、9、10或更多倍之任一者)體內半衰期。In some embodiments, the multispecific constructs described herein have increased (e.g., at least about 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more fold) in vivo half-life compared to the anti-IL-13 antibody portion (e.g., Fab, scFv) alone.

在一些實施例中，該抗IL-13抗體部分(例如，全長、sdAb、scFv或Fab)以≤ 1 μM，諸如≤100 nM，較佳為≤10 nM，更佳為≤1 nM的Kd結合至IL-13抗原。舉例而言，該抗IL-13抗體部分之Kd值係介於約1 nM與約1 pM之間。在一些實施例中，該抗IL-13抗體部分以約1×10-11M至約1×10-7M(例如，約1×10-11M至約1×10-10M、約1×10-11M至約1×10-9M、約1×10-11M至約1×10-8M、約1×10-10M至約1×10-9M、約1×10-10M至約1×10-8M、約1×10-10M至約1×10-7M、約1×10-9M至約1×10-7M、約1×10-9M至約1×10-8M，或約1×10-8M至約1×10-7M)的Kd結合至人類IL-13及/或猴(例如，食蟹獼猴) IL-13。In some embodiments, the anti-IL-13 antibody portion (e.g., full length, sdAb, scFv, or Fab) binds to the IL-13 antigen with aKd of ≤ 1 μM, such as ≤ 100 nM, preferably ≤ 10 nM, and more preferably ≤ 1 nM. For example, the anti-IL-13 antibody portion has aKd value between about 1 nM and about 1 pM. In some embodiments, the anti-IL-13 antibody portion binds to human IL-13 and/or monkey (e.g., cynomolgus macaque)IL-13 with a Kd of about 1×10-11 M to about 1×10-10 M, about 1×10-11 M to about 1×10-9 M, about1×10-11M to about 1×10-8 M, about 1×10-10 M to about 1×10-9 M, about 1×10-10 M toabout 1×10-8 M, about 1×10-10 M to about 1×10-7 M, about 1×10-9 M to about 1×10-7 M, about 1×10-9M to about 1×10-8 M, or about 1×10-8 M to about 1×10-7 M.

本文所述之抗IL-13抗體部分可為任何形式並衍生自任何合適的抗IL-13抗體或其抗原結合片段。舉例而言，該抗IL-13抗體部分可選自全長抗體、scFv、VH、VL、scFv-scFv、Fv、Fab、Fab’、(Fab’)2、微型抗體(minibody)、雙抗體(diabody,)、域抗體變體(domain antibody variant，dAb)、單域抗體(single domain antibody，sdAb)、駱駝科抗體(VHH)、纖維接合素3域變體(ibronectin 3 domain variant)、錨蛋白重複變體(ankyrin repeat variant)及其他衍生自其他蛋白質框架的抗原特異性結合域。在一些實施例中，該抗IL-13抗體部分為Fab。在一些實施例中，該抗IL-13抗體部分為scFv。在一些實施例中，該抗IL-13抗體部分係人源化。在一些實施例中，該抗IL-13抗體部分係嵌合。在一些實施例中，該抗IL-13抗體部分係衍生自小鼠、大鼠、猴或兔的單株抗體。在一些實施例中，該抗IL-13抗體部分係衍生自本領域已知之任何抗IL-13單株抗體。在一些實施例中，該抗IL-13抗體部分係衍生自完整人類抗體，例如使用噬菌體展示、酵母菌展示或帶有人類Ig基因的轉基因小鼠開發。在一些實施例中，該抗IL-13抗體部分為單特異性抗體、多特異性抗體、單價抗體或多價(例如，雙價)抗體。The anti-IL-13 antibody portion described herein can be in any form and derived from any suitable anti-IL-13 antibody or antigen-binding fragment thereof. For example, the anti-IL-13 antibody portion can be selected from a full-length antibody, scFv, VH, VL, scFv-scFv, Fv, Fab, Fab', (Fab')2 , minibody, diabody, domain antibody variant (dAb), single domain antibody (sdAb), camelid antibody (VHH), ibronectin 3 domain variant, ankyrin repeat variant, and other antigen-specific binding domains derived from other protein frameworks. In some embodiments, the anti-IL-13 antibody portion is a Fab. In some embodiments, the anti-IL-13 antibody portion is a scFv. In some embodiments, the anti-IL-13 antibody portion is humanized. In some embodiments, the anti-IL-13 antibody portion is chimeric. In some embodiments, the anti-IL-13 antibody portion is derived from a monoclonal antibody of mouse, rat, monkey, or rabbit. In some embodiments, the anti-IL-13 antibody portion is derived from any anti-IL-13 monoclonal antibody known in the art. In some embodiments, the anti-IL-13 antibody portion is derived from a fully human antibody, for example, developed using phage display, yeast display, or transgenic mice carrying human Ig genes. In some embodiments, the anti-IL-13 antibody portion is a monospecific antibody, a multispecific antibody, a monovalent antibody, or a multivalent (e.g., bivalent) antibody.

在一些實施例中，該抗IL-13抗體部分特異性地結合至人類及非人類靈長類動物(諸如食蟹獼猴) IL-13。在一些實施例中，該抗IL-13抗體部分僅結合至人類IL-13。示例性抗IL-13抗體及其抗原結合片段包括但不限於Adbry™ (曲羅蘆單抗-ldrm)、來瑞珠單抗及以下揭露的內容：US20220177565；US 9,394,361；US 7,585,500；US 8,221,752；US 7,915,388；US 8,399,630；US 8,323,646；US 7,910,708；US 7,807,788；US 7,615,213；US 7,501,121；US20070048785；US 7,674,459；US 11,434,286；US 8,318,160；US 9,067,994；US 10,597,447；US 7,829,090；US 7,910,708；US 11,136,387；US 7,947,273；US 9,120,870；US 11,344,621；WO2022157773；US20210087284；WO2005062967；WO2015127405；US 8,388,965；及US 8,691,233，其各自之內容皆通過引用具體併入本文中。採用與猴IL-13具有交叉反應性的此類抗IL-13抗體部分的多特異性構築體或分離的抗IL-13抗體構築體可有助於非人類靈長類動物的毒性研究，其可為人類臨床試驗候選者提供更相關的安全性評估，而無需在黑猩猩身上進行毒性研究或使用替代分子。在一些實施例中，該抗IL-13抗體部分與人類IL-13的結合比非人類(例如，食蟹獼猴) IL-13的更強(諸如至少約1.5、2、5、10、20、50、100、1000或更多倍之任一者)。In some embodiments, the anti-IL-13 antibody portion specifically binds to human and non-human primate (e.g., cynomolgus macaque) IL-13. In some embodiments, the anti-IL-13 antibody portion binds only to human IL-13. Exemplary anti-IL-13 antibodies and antigen-binding fragments thereof include, but are not limited to, Adbry™ (trorazolomumab-ldrm), lerezumab, and those disclosed in US20220177565; US 9,394,361; US 7,585,500; US 8,221,752; US 7,915,388; US 8,399,630; US 8,323,646; US 7,910,708; US 7,807,788; US 7,615,213; US 7,501,121; US20070048785; US 7,674,459; US 11,434,286; US 8,318,160; US 9,067,994; US 10,597,447; US 7,829,090; US 7,910,708; US 11,136,387; US 7,947,273; US 9,120,870; US 11,344,621; WO2022157773; US20210087284; WO2005062967; WO2015127405; US 8,388,965; and US 8,691,233, the contents of each of which are specifically incorporated herein by reference. Multispecific constructs or isolated anti-IL-13 antibody constructs employing such anti-IL-13 antibody portions that are cross-reactive with monkey IL-13 can facilitate toxicity studies in non-human primates, which can provide more relevant safety assessments for human clinical trial candidates without the need for toxicity studies in chimpanzees or the use of surrogate molecules. In some embodiments, the anti-IL-13 antibody portion binds to human IL-13 more strongly (e.g., at least about 1.5, 2, 5, 10, 20, 50, 100, 1000, or more) than to non-human (e.g., cynomolgus macaque) IL-13.

抗IL-13抗體部分可完全或部分調控、阻斷、抑制、降低、拮抗、中和或干擾IL-13的功能活性。在抗IL-13抗體部分存在下，當IL-13的功能活性降低至少約95% (諸如至少約96%、97%、98%、99%或100%之任一者)時，與不被抗IL-13抗體部分結合時相比，該抗IL-13抗體部分被視為能夠完全調控、阻斷、抑制、降低、拮抗、中和或干擾IL-13的功能活性。在抗IL-13抗體部分存在下，當IL-13的功能活性降低至少約50% (諸如至少約55%、60%、70%、75%、80%、85%或90%之任一者)時，與不被抗IL-13抗體部分結合時相比，該抗IL-13抗體部分被視為能夠顯著調控、阻斷、抑制、降低、拮抗、中和或干擾IL-13的功能活性。在抗IL-13抗體部分存在下，當標靶抗原的功能活性降低小於約50% (諸如降低小於約10%、15%、20%、25%、30%、35%、或40%或45%之任一者)時，與不被抗IL-13抗體部分結合時相比，該抗IL-13抗體部分被視為能夠部分調控、阻斷、抑制、降低、拮抗、中和或干擾IL-13的功能活性。The anti-IL-13 antibody portion can fully or partially modulate, block, inhibit, reduce, antagonize, neutralize, or interfere with the functional activity of IL- 13. When the functional activity of IL-13 is reduced by at least about 95% (e.g., at least about any of 96%, 97%, 98%, 99%, or 100%) in the presence of the anti-IL-13 antibody portion compared to when not bound by the anti-IL-13 antibody portion, the anti-IL-13 antibody portion is considered capable of fully modulating, blocking, inhibiting, reducing, antagonizing, neutralizing, or interfering with the functional activity of IL-13. When the functional activity of IL-13 is reduced by at least about 50% (e.g., at least about any of 55%, 60%, 70%, 75%, 80%, 85%, or 90%) in the presence of the anti-IL-13 antibody portion compared to when not bound by the anti-IL-13 antibody portion, the anti-IL-13 antibody portion is considered capable of significantly modulating, blocking, inhibiting, reducing, antagonizing, neutralizing, or interfering with the functional activity of IL-13. When the functional activity of the target antigen is reduced by less than about 50% (e.g., by less than about 10%, 15%, 20%, 25%, 30%, 35%, or 40% or 45%) in the presence of the anti-IL-13 antibody portion compared to when not bound by the anti-IL-13 antibody portion, the anti-IL-13 antibody portion is considered capable of partially modulating, blocking, inhibiting, reducing, antagonizing, neutralizing, or interfering with the functional activity of IL-13.

在一些實施例中，該抗IL-13抗體部分在序列中含有一或多個變異。在一些實施例中，該變體序列中的胺基酸變異(例如，取代)不會實質上降低 (例如，降低至多約50%、40%、30%、20%、10%、5%、1%或更少之任一者)抗IL-13抗體部分結合至IL-13的能力。亦考慮實質上改善(例如，改善至少約1.2、1.5、2、5、10、20、50 或更多倍之任一者)抗IL-13抗體部分與IL-13之結合親和力或其他性質(諸如特異性、免疫原性及/或與IL-13表位變體的交叉反應)的修飾。In some embodiments, the anti-IL-13 antibody portion contains one or more variations in sequence. In some embodiments, the amino acid variations (e.g., substitutions) in the variant sequence do not substantially reduce (e.g., by at most about 50%, 40%, 30%, 20%, 10%, 5%, 1%, or less) the ability of the anti-IL-13 antibody portion to bind to IL-13. Modifications that substantially improve (e.g., by at least about 1.2, 1.5, 2, 5, 10, 20, 50, or more fold) the binding affinity or other properties of the anti-IL-13 antibody portion to IL-13, such as specificity, immunogenicity, and/or cross-reactivity with IL-13 epitope variants, are also contemplated.

在一些實施例中，該抗IL-13抗體部分為scFv (本文中亦稱為「抗IL-13 scFv」)。在一些實施例中，該抗IL-13 scFv之N端至C端包含：VH-視情況的連接子-VL，或VL-視情況的連接子-VH。以下「連接子」小節中的任何連接子皆可用於連接抗IL-13 scFv的VH及VL。In some embodiments, the anti-IL-13 antibody portion is a scFv (also referred to herein as an "anti-IL-13 scFv"). In some embodiments, the anti-IL-13 scFv comprises, from the N-terminus to the C-terminus, VH-optional linker-VL, or VL-optional linker-VH. Any linker described below in the "Linker" section can be used to link the VH and VL of the anti-IL-13 scFv.

在一些實施例中，該抗IL-13抗體部分為Fab片段(本文中亦稱為「抗IL-13 Fab」或「抗IL-13 Fab片段」)，其包含含有VH及CH1的第一多肽，以及含有VL及CL的第二多肽。在一些實施例中，該抗IL-13 Fab片段內之可變區及恆定區的配置可能與天然抗IL-13 Fab片段中發現的不同。舉例而言，在一些實施例中，該抗IL-13 Fab片段包含含有VH及CL的第一多肽，以及含有VL及CH1的第二多肽(參見例如Shaefer等人(2011)，PNAS，108:111870-92，其內容通過引用整體併入本文)。In some embodiments, the anti-IL-13 antibody portion is a Fab fragment (also referred to herein as an "anti-IL-13 Fab" or "anti-IL-13 Fab fragment") comprising a first polypeptide comprising VH and CH1, and a second polypeptide comprising VL and CL. In some embodiments, the configuration of the variable and constant regions within the anti-IL-13 Fab fragment may differ from that found in native anti-IL-13 Fab fragments. For example, in some embodiments, the anti-IL-13 Fab fragment comprises a first polypeptide comprising VH and CL, and a second polypeptide comprising VL and CH1 (see, e.g., Shaefer et al. (2011), PNAS, 108:111870-92, the contents of which are incorporated herein by reference in their entirety).

在一些實施例中，該抗IL-13 Fab中的CH1及VH與VL及CL進行異二聚化，並藉由重鏈與輕鏈恆定區之間的雙硫鍵共價連接。在一些實施例中，該抗IL-13 Fab片段具有基本結構NH2-VL-CL-S-S-CH1-VH-NH2。在一些實施例中，該抗IL-13 Fab的CH1及CL藉由一或多個雙硫鍵連接。在一些實施例中，該抗IL-13 Fab片段之CH1與CL之間的雙硫鍵數量為至少1個，諸如2、3、4、5個或更多。在一些實施例中，該抗IL-13 Fab片段(諸如CH1及CL區域)中的半胱胺酸殘基係經工程改造以導入雙硫鍵。In some embodiments, the CH1 and VH of the anti-IL-13 Fab heterodimerize with the VL and CL and are covalently linked by disulfide bonds between the heavy and light chain constant regions. In some embodiments, the anti-IL-13 Fab fragment has the basic structure NH2-VL -CL-SS-CH1-VH-NH2 . In some embodiments, the CH1 and CL of the anti-IL-13 Fab are linked by one or more disulfide bonds. In some embodiments, the number of disulfide bonds between the CH1 and CL of the anti-IL-13 Fab fragment is at least one, such as two, three, four, five, or more. In some embodiments, cysteine residues in the anti-IL-13 Fab fragment (e.g., CH1 and CL regions) are engineered to introduce disulfide bonds.

在一些實施例中，該抗IL-13 Fab片段在C端處不含雙硫鍵。舉例而言，該抗IL-13 Fab片段之重鏈及輕鏈可以此方式進行工程改造，以便穩定相互作用而不需要雙硫鍵。舉例而言，重鏈及輕鏈可以此方式進行工程改造，以便穩定相互作用而不需要雙硫鍵。在一些實施例中，重鏈或輕鏈可經工程改造以移除半胱胺酸殘基，且其中重鏈及輕鏈仍然穩定地相互作用並發揮Fab的功用。在一些實施例中，進行突變以促進重鏈與輕鏈之間的穩定相互作用。舉例而言，「杵臼」工程改造策略可用於促進Fab的重鏈與輕鏈之間的二聚化(參見例如1996 Protein Engineering，9:617 - 621)。本文亦考慮使用針對特定目的而設計的變體Fab片段，例如CH1及/或CL之恆定域的胺基酸變化，以及移除雙硫鍵或添加用於純化的標籤等。在以下「恆定域及Fc域」小節中更詳細地描述此等突變。In some embodiments, the anti-IL-13 Fab fragment does not contain a disulfide bond at the C-terminus. For example, the heavy and light chains of the anti-IL-13 Fab fragment can be engineered to stabilize their interaction without the need for a disulfide bond. For example, the heavy and light chains can be engineered to stabilize their interaction without the need for a disulfide bond. In some embodiments, the heavy or light chain can be engineered to remove a cysteine residue, while still allowing the heavy and light chains to stably interact and function as a Fab. In some embodiments, mutations are made to promote a stable interaction between the heavy and light chains. For example, a "knob-in-hole" engineering strategy can be used to promote dimerization between the heavy and light chains of a Fab (see, e.g., Protein Engineering, 1996, 9:617-621). This article also contemplates the use of variant Fab fragments designed for specific purposes, such as amino acid changes in the homeostasis domains of CH1 and/or CL, removal of disulfide bonds, or addition of tags for purification. These mutations are described in more detail in the "Homeostasis Domain and Fc Domain" section below.

在一些實施例中，該抗IL-13 Fab片段之CH1及CL由約2個雙硫鍵連接。在一些實施例中，該抗IL-13 Fab片段包含人類免疫球蛋白CH1，例如含有SEQ ID NO:158-162之任一者之胺基酸序列。在一些實施例中，該抗IL-13 Fab片段包含人類λ輕鏈恆定區，例如含有SEQ ID NO:74或75之胺基酸序列。在一些實施例中，該抗IL-13 Fab片段包含人類κ輕鏈恆定區，例如含有SEQ ID NO:73之胺基酸序列。In some embodiments, the CH1 and CL of the anti-IL-13 Fab fragment are linked by approximately two disulfide bonds. In some embodiments, the anti-IL-13 Fab fragment comprises a human immunoglobulin CH1, e.g., comprising the amino acid sequence of any one of SEQ ID NOs: 158-162. In some embodiments, the anti-IL-13 Fab fragment comprises a human lambda light chain constant region, e.g., comprising the amino acid sequence of SEQ ID NOs: 74 or 75. In some embodiments, the anti-IL-13 Fab fragment comprises a human kappa light chain constant region, e.g., comprising the amino acid sequence of SEQ ID NO: 73.

在一些實施例中，該抗IL-13抗體部分(例如，scFv或Fab)特異性地辨識IL-13。在一些實施例中，該抗IL-13抗體部分包含含有選自SEQ ID NO:65、151、153、155及157之任一者之一、二或三個CDR的VH及/或含有選自SEQ ID NO:69、114、118、147、149及191-196之任一者之一、二或三個CDR的VL。在一些實施例中，該抗IL-13抗體部分包含含有選自SEQ ID NO:65、151、153、155及157之任一者之三個CDR的VH及/或含有選自SEQ ID NO:69、114、118、147、149及191-196之任一者之三個CDR的VL。在一些實施例中，該抗IL-13抗體部分包含：含有選自SEQ ID NO:66-68、150、152、154及156之任一者之一、二或三個CDR的VH及/或含有選自SEQ ID NO:70-72、115、119、146、148及191-196之任一者之一、二或三個CDR的VL。In some embodiments, the anti-IL-13 antibody portion (e.g., scFv or Fab) specifically recognizes IL- 13. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising one, two, or three CDRs selected from any one of SEQ ID NOs: 65, 151, 153, 155, and 157, and/or a VL comprising one, two, or three CDRs selected from any one of SEQ ID NOs: 69, 114, 118, 147, 149, and 191-196. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising three CDRs selected from any one of SEQ ID NOs: 65, 151, 153, 155, and 157 and/or a VL comprising three CDRs selected from any one of SEQ ID NOs: 69, 114, 118, 147, 149, and 191-196. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising one, two, or three CDRs selected from any one of SEQ ID NOs: 66-68, 150, 152, 154, and 156 and/or a VL comprising one, two, or three CDRs selected from any one of SEQ ID NOs: 70-72, 115, 119, 146, 148, and 191-196.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:70之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:70之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體及含有SEQ ID NO:69之胺基酸序列的VL或其與SEQ ID NO:69之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 69.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:115之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:115之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:114之胺基酸序列的VL或其與SEQ ID NO:114之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:114之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 115 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 115, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 114, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 114.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:119之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:119之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:118之胺基酸序列的VL或其與SEQ ID NO:118之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:118之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 119, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 119, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 118, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 118. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 118.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:146之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:146之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:147之胺基酸序列的VL或其與SEQ ID NO:147之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:147之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 146, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 146, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 147, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 147. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 147.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:148之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:148之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，諸如保留性取代)，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:149之胺基酸序列的VL或其與SEQ ID NO:149之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:149之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 148, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 148, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71 (e.g., conservative substitutions), and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 149, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 149. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 149.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:150之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:115之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:150之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:115之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:151之胺基酸序列的VH或其與SEQ ID NO:151之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:118之胺基酸序列的VL或其與SEQ ID NO:114之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:151之胺基酸序列的VH及含有SEQ ID NO:114之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 150, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 115 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 150, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 115, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 151, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 151, and a VL comprising the amino acid sequence of SEQ ID NO: 118, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 151, and a VL comprising the amino acid sequence of SEQ ID NO: 114.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:152之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:115之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:152之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:115之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:153之胺基酸序列的VH或其與SEQ ID NO:153之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:114之胺基酸序列的VL或其與SEQ ID NO:114之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:153之胺基酸序列的VH，以及含有SEQ ID NO:114之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 152, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 115 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 152, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 115, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 153, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 153, and a VL comprising the amino acid sequence of SEQ ID NO: 114, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 153, and a VL comprising the amino acid sequence of SEQ ID NO: 114.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:154之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:115之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:154之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:115之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:155之胺基酸序列的VH或其與SEQ ID NO:155之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:114之胺基酸序列的VL或其與SEQ ID NO:114之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:155之胺基酸序列的VH及含有SEQ ID NO:114之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 154, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 115 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 154, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 115, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 155, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 155, and a VL comprising the amino acid sequence of SEQ ID NO: 114, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 155, and a VL comprising the amino acid sequence of SEQ ID NO: 114.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:156之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:115之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:156之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:115之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:157之胺基酸序列的VH或其與SEQ ID NO:157之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:114之胺基酸序列的VL或其與SEQ ID NO:114之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:157之胺基酸序列的VH及含有SEQ ID NO:114之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 156, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 115 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 156, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 115, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 157, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 157, and a VL comprising the amino acid sequence of SEQ ID NO: 114, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 114. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 157 and a VL comprising the amino acid sequence of SEQ ID NO: 114.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:182之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:182之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:191之胺基酸序列的VL或其與SEQ ID NO:191之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:191之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 182 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 182, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 191, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 191. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 191.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:183之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:183之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:192之胺基酸序列的VL或其與SEQ ID NO:192之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:192之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 183 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 183, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 192, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 192. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 192.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:184之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:184之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:193之胺基酸序列的VL或其與SEQ ID NO:193之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:193之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 184 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 184, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 193, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 193. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 193.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:185之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:185之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:194之胺基酸序列的VL或其與SEQ ID NO:194之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:194之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 185 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 185, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 194, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 194. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 194.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:186之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:186之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:195之胺基酸序列的VL或其與SEQ ID NO:195之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:195之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 186, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 186, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 195, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 195. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 195.

在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:187之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體、含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:66之胺基酸序列的CDR-H1、含有SEQ ID NO:67之胺基酸序列的CDR-H2、含有SEQ ID NO:68之胺基酸序列的CDR-H3、含有SEQ ID NO:187之胺基酸序列的CDR-L1、含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及含有SEQ ID NO:72之胺基酸序列的CDR-L3。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH或其與SEQ ID NO:65之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體，以及含有SEQ ID NO:196之胺基酸序列的VL或其與SEQ ID NO:196之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:196之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H4 comprising the amino acid sequence of SEQ ID NO: 69, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H5 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions), a CDR-H6 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to about 3 NO: 187 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); a CDR-L2 containing the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and a CDR-L3 containing the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the anti-IL-13 antibody portion comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 187, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 196, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 196. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65, and a VL comprising the amino acid sequence of SEQ ID NO: 196.

在一些實施例中，該抗IL-13抗體部分包含含有來自SEQ ID NO:65之一、二或三個CDR的VH及/或含有來自SEQ ID NO:69之一、二或三個CDR的VL。在一些實施例中，該抗IL-13抗體部分包含含有來自SEQ ID NO:65之三個CDR的VH及/或含有來自SEQ ID NO:69之三個CDR的VL。在一些實施例中，該抗IL-13抗體部分包含：含有選自SEQ ID NO:66-68之任一者之一、二或三個CDR的VH及/或含有選自SEQ ID NO:70-72之任一者之一、二或三個CDR的VL。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及/或含有SEQ ID NO:69之胺基酸序列的VL。在一些實施例中，該抗IL-13抗體部分包含含有SEQ ID NO:65之胺基酸序列的VH及含有SEQ ID NO:69之胺基酸序列的VL。In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising one, two, or three CDRs from SEQ ID NO: 65 and/or a VL comprising one, two, or three CDRs from SEQ ID NO: 69. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising three CDRs from SEQ ID NO: 65 and/or a VL comprising three CDRs from SEQ ID NO: 69. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising one, two, or three CDRs selected from any one of SEQ ID NOs: 66-68 and/or a VL comprising one, two, or three CDRs selected from any one of SEQ ID NOs: 70-72. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65 and/or a VL comprising the amino acid sequence of SEQ ID NO: 69. In some embodiments, the anti-IL-13 antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 65 and a VL comprising the amino acid sequence of SEQ ID NO: 69.

在一些實施例中，該抗IL-13抗體部分與參考抗IL-13抗體競爭結合(例如，競爭結合至少約50%、55%、60%、70%、80%、90%、95%、96%、97%、98%、99%或更多之一者)至IL-13，該參考抗IL-13抗體包含(a)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:69之胺基酸序列的參考VL；(b)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:114之胺基酸序列的參考VL；(c)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:118之胺基酸序列的參考VL；(d)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:147之胺基酸序列的參考VL；(e)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:149之胺基酸序列的參考VL；(f)含有SEQ ID NO:151之胺基酸序列的參考VH及含有SEQ ID NO:114之胺基酸序列的參考VL；(g)含有SEQ ID NO:153之胺基酸序列的參考VH及含有SEQ ID NO:114之胺基酸序列的參考VL；(h)含有SEQ ID NO:155之胺基酸序列的參考VH及含有SEQ ID NO:114之胺基酸序列的參考VL；(i)含有SEQ ID NO:157之胺基酸序列的參考VH及含有SEQ ID NO:114之胺基酸序列的參考VL；(j)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:191之胺基酸序列的參考VL；(k)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:192之胺基酸序列的參考VL；(l)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:193之胺基酸序列的參考VL；(m)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:194之胺基酸序列的參考VL；(n)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:195之胺基酸序列的參考VL；或(o)含有SEQ ID NO:65之胺基酸序列的參考VH及含有SEQ ID NO:196之胺基酸序列的參考VL。在一些實施例中，提供含有VH及VL的抗IL-13抗體部分，其中：(a)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:69之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(b)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:114之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(c)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:118之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(d)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:147之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(e)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:149之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(f)該VH包含含有SEQ ID NO:151之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:114之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(g)該VH包含含有SEQ ID NO:153之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:114之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(h)該VH包含含有SEQ ID NO:155之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:114之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(i)該VH包含含有SEQ ID NO:157之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:114之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(j)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:191之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(k)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:192之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(l)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:193之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(m)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:194之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；(n)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:195之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列；或(o)該VH包含含有SEQ ID NO:65之胺基酸序列的參考VH的CDR-H1、CDR-H2及CDR-H3序列，且該VL包含含有SEQ ID NO:196之胺基酸序列的參考VL的CDR-L1、CDR-L2及CDR-L3序列。In some embodiments, the anti-IL-13 antibody portion competes for binding (e.g., competes for at least about 50%, 55%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or more) to IL-13 with a reference anti-IL-13 antibody comprising (a) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 69; (b) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (c) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 118; (d) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: NO: 147; (e) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 149; (f) a reference VH comprising the amino acid sequence of SEQ ID NO: 151 and a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (g) a reference VH comprising the amino acid sequence of SEQ ID NO: 153 and a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (h) a reference VH comprising the amino acid sequence of SEQ ID NO: 155 and a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (i) a reference VH comprising the amino acid sequence of SEQ ID NO: 157 and a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (j) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: NO: 191; (k) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 192; (l) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 193; (m) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 194; (n) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 195; or (o) a reference VH comprising the amino acid sequence of SEQ ID NO: 65 and a reference VL comprising the amino acid sequence of SEQ ID NO: 196. In some embodiments, an anti-IL-13 antibody portion comprising a VH and a VL is provided, wherein: (a) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 69; (b) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (c) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 114 NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 118; (d) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 147; (e) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-H2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: (f) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 151, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (g) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 153, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (h) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 154. NO: 155, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (i) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 157, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 114; (j) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: NO: 191; (k) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 192; (l) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 193; (m) the VH comprises the CDR-H1, CDR-H2, and CDR-L3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 194; (n) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: 195; or (o) the VH comprises the CDR-H1, CDR-H2, and CDR-H3 sequences of a reference VH comprising the amino acid sequence of SEQ ID NO: 65, and the VL comprises the CDR-L1, CDR-L2, and CDR-L3 sequences of a reference VL comprising the amino acid sequence of SEQ ID NO: The amino acid sequence of NO:196 is based on the CDR-L1, CDR-L2, and CDR-L3 sequences of VL.

在一些實施例中，該抗IL-13抗體部分為scFv (「抗IL-13 scFv」)。在一些實施例中，該抗IL-13 scFv包含經由視情況的連接子彼此融合的VH及VL。在一些實施例中，該視情況的連接子包含選自由以下所組成群組之胺基酸序列：GG及SEQ ID NO:14、16-18、97-99、163-172及224，諸如GG及SEQ ID NO:98、99及224之任一者。在一些實施例中，該抗IL-13 scFv含有SEQ ID NO:108之胺基酸序列或其與SEQ ID NO:108之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗 IL-13 scFv含有SEQ ID NO:108之胺基酸序列。In some embodiments, the anti-IL-13 antibody portion is a scFv ("anti-IL-13 scFv"). In some embodiments, the anti-IL-13 scFv comprises a VH and a VL fused to each other via an optional linker. In some embodiments, the optional linker comprises an amino acid sequence selected from the group consisting of GG and SEQ ID NOs: 14, 16-18, 97-99, 163-172, and 224, such as GG and any one of SEQ ID NOs: 98, 99, and 224. In some embodiments, the anti-IL-13 scFv comprises the amino acid sequence of SEQ ID NO: 108, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 108. In some embodiments, the anti-IL-13 scFv comprises the amino acid sequence of SEQ ID NO: 108.

在一些實施例中，該抗IL-13抗體部分為Fab片段 (「抗IL-13 Fab」)。在一些實施例中，該抗IL-13 Fab片段包含含有SEQ ID NO:124之胺基酸序列的第一多肽或其與SEQ ID NO:124之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體；及含有SEQ ID NO:102或197之胺基酸序列的第二多肽或其與SEQ ID NO:102或197之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗IL-13 Fab片段包含含有SEQ ID NO:124之胺基酸序列的第一多肽及含有SEQ ID NO:102或197之胺基酸序列的第二多肽。In some embodiments, the anti-IL-13 antibody portion is a Fab fragment ("anti-IL-13 Fab"). In some embodiments, the anti-IL-13 Fab fragment comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 124; and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 102 or 197. In some embodiments, the anti-IL-13 Fab fragment comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124, and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197.

在一些實施例中，該抗IL-13抗體部分為全長抗體(「抗IL-13全長抗體」)。在一些實施例中，該抗IL-13全長抗體包含人類κ CL或人類λ CL，諸如含有SEQ ID NO:73-75之任一者之胺基酸序列的CL。在一些實施例中，該抗IL-13全長抗體包含衍生自人類IgG1、IgG2、IgG3或IgG4 (諸如人類IgG1)的Fc域。以下「Fc域」小節中所述之任何Fc域皆可在此使用。在一些實施例中，該Fc域包含：i) L234A+L235A (「LALA」)突變，ii) M252Y+S254T+T256E (「YTE」)突變，及/或iii) M428L+N434S (「LS」)突變，其係根據EU編號。在一些實施例中，該Fc域進一步包含杵臼突變，諸如在一個Fc次單元中的T366W 「杵」突變，以及在另一個Fc次單元中的T366S+L368A+Y407V 「臼」突變。在一些實施例中，該抗IL-13全長抗體包含：i)該Fc域之第一次單元及/或第二次單元，其各自含有SEQ ID NO:76-79之任一者的胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；ii)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；iii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；iv)該Fc域之第一次單元含有SEQ ID NO:95之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:96之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；或v)該Fc域之第一次單元含有SEQ ID NO:96之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:95之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段。在一些實施例中，該抗IL-13全長抗體包含：i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；ii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；iii)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；iv)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；v)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈；vi)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈；vii)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；viii)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈；ix)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或x)含有SEQ ID NO:130之胺基酸序列的第一重鏈、含有SEQ ID NO:131之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。In some embodiments, the anti-IL-13 antibody portion is a full-length antibody ("anti-IL-13 full-length antibody"). In some embodiments, the anti-IL-13 full-length antibody comprises human kappa CL or human lambda CL, such as a CL comprising the amino acid sequence of any one of SEQ ID NOs: 73-75. In some embodiments, the anti-IL-13 full-length antibody comprises an Fc domain derived from human IgG1, IgG2, IgG3, or IgG4 (such as human IgG1). Any Fc domain described below in the "Fc Domain" section can be used herein. In some embodiments, the Fc domain comprises: i) L234A+L235A ("LALA") mutations, ii) M252Y+S254T+T256E ("YTE") mutations, and/or iii) M428L+N434S ("LS") mutations, according to EU numbering. In some embodiments, the Fc domain further comprises a knob-hole mutation, such as a T366W "knob" mutation in one Fc subunit and a T366S+L368A+Y407V "hole" mutation in the other Fc subunit. In some embodiments, the anti-IL-13 full-length antibody comprises: i) the first and/or second Fc domain units, each of which comprises the amino acid sequence of any one of SEQ ID NOs: 76-79 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; ii) the first Fc domain unit comprises the amino acid sequence of SEQ ID NO: 80 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215, and the second Fc domain unit comprises the amino acid sequence of SEQ ID NO: 81 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; iii) the first Fc domain unit comprises the amino acid sequence of SEQ ID NO: 81 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215, and the second Fc domain unit comprises the amino acid sequence of SEQ ID NO: 80 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; iv) the first Fc domain unit comprises the amino acid sequence of SEQ ID NO: 95 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 96. NO: 215, and the second Fc domain subunit contains the amino acid sequence of SEQ ID NO: 96 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; or v) the first Fc domain subunit contains the amino acid sequence of SEQ ID NO: 96 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215, and the second Fc domain subunit contains the amino acid sequence of SEQ ID NO: 95 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215. In some embodiments, the anti-IL-13 full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 101 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; iii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 123 and two light chains each comprising the amino acid sequence of SEQ ID NO: 121 or 206; iv) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 209 and two light chains each comprising the amino acid sequence of SEQ ID NO: 122 or 207; v) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 101 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197; NO: 210 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207; vi) two heavy chains each containing the amino acid sequence of SEQ ID NO: 211 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206; vii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 212 and two light chains each containing the amino acid sequence of SEQ ID NO: 120 or 208; viii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 213 and two light chains each containing the amino acid sequence of SEQ ID NO: 120 or 208; ix) two heavy chains each containing the amino acid sequence of SEQ ID NO: 225 and two light chains each containing the amino acid sequence of SEQ ID NO: or x) a first heavy chain comprising the amino acid sequence of SEQ ID NO: 130, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 131, and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197.

在一些實施例中，本文所述之分離的抗IL-13抗體構築體之任一者之一或多個多肽的N端係額外與訊號肽融合以更好地表現。In some embodiments, the N-terminus of one or more polypeptides of any of the isolated anti-IL-13 antibody constructs described herein is additionally fused to a signal peptide for better expression.

在一些實施例中，本文所述之分離的抗IL-13抗體構築體之任一者之一或多個多肽的N端及/或C端可含有組胺酸標籤(HIS-標籤)以進行蛋白質純化。舉例而言，該抗IL-13全長抗體之一或多個多肽的C端可進一步包含組胺酸標籤。額外的抗體部分In some embodiments, one or more polypeptides of any of the isolated anti-IL-13 antibody constructs described herein may contain a histidine tag (HIS-tag) at the N-terminus and/or C-terminus for protein purification. For example, the C-terminus of one or more polypeptides of the full-length anti-IL-13 antibody may further contain a histidine tag.Additional Antibody Moieties

特異性地結合至如本文所述之第二、第三或第四抗原的抗體部分之任一者可用作本文所述之多特異性構築體之任一者的第二、第三或第四抗體部分。在一些實施例中，第二、第三或第四抗原係選自由以下組成之群組：TSLP、IL-17A、IL-17F、TNF-α及IFN-γ。抗TSLP抗體部分Any of the antibody moieties that specifically bind to a second, third, or fourth antigen as described herein can be used as the second, third, or fourth antibody moiety of any of the multispecific constructs described herein. In some embodiments, the second, third, or fourth antigen is selected from the group consisting of TSLP, IL-17A, IL-17F, TNF-α, and IFN-γ.Anti-TSLPAntibody Moiety

本文所述之抗TSLP抗體部分之任一者可用作本文所述之多特異性構築體之任一者的第二、第三或第四抗體部分。Any of the anti-TSLP antibody portions described herein can be used as the second, third, or fourth antibody portion of any of the multispecific constructs described herein.

TSLP為由腸道及肺上皮細胞、皮膚角質細胞及樹突細胞產生的細胞激素，但其亦可由(例如)呼吸道平滑肌細胞、肥大細胞、單核球、巨噬細胞、顆粒細胞、滑液膜纖維母細胞(synovial fibroblast)等產生。TSLP涉及Th2發炎反應，且已被確定為治療氣喘的可能的治療標靶。此示例性抗原可用作特定發炎性疾病(例如，過敏或自體免疫疾病)發展的生物標記、預後指標及抗原表現發炎性疾病的免疫治療標靶。TSLP is a cytokine produced by intestinal and lung epithelial cells, skin keratinocytes, and dendritic cells. It is also produced by, for example, airway smooth muscle cells, mast cells, monocytes, macrophages, granulocytes, synovial fibroblasts, and others. TSLP is involved in Th2 inflammatory responses and has been identified as a potential therapeutic target for the treatment of asthma. This exemplary antigen can be used as a biomarker for the development of specific inflammatory diseases (e.g., allergies or autoimmune diseases), a prognostic indicator, and as a target for immunotherapy of antigen-expressing inflammatory diseases.

本文所述之多特異性構築體(諸如雙特異性構築體)可包含一或多個(例如，1、2、3、4、5或多個，諸如1或2個)特異性地結合至TSLP的抗TSLP抗體部分(例如，全長抗體、scFv、Fab)。在一些實施例中，該多特異性構築體包含二或多個抗TSLP抗體部分(例如，scFv，例如Fab)。The multispecific constructs described herein (e.g., bispecific constructs) can comprise one or more (e.g., 1, 2, 3, 4, 5, or more, such as 1 or 2) anti-TSLP antibody portions (e.g., full-length antibodies, scFv, Fab) that specifically bind to TSLP. In some embodiments, the multispecific constructs comprise two or more anti-TSLP antibody portions (e.g., scFv, such as Fab).

TSLP抗原可來自各種動物物種，包括人類、靈長類、小鼠、大鼠、兔或其他哺乳動物。在一些實施例中，該TSLP分子為人類TSLP。在一些實施例中，該TSLP分子為大鼠TSLP。在一些實施例中，該TSLP分子為食蟹獼猴TSLP。在一些實施例中，該TSLP分子為野生型。在一些實施例中，該TSLP分子為突變體(例如，與來自同一物種的野生型TSLP序列相比，包含一或多個插入、缺失及/或胺基酸取代)。TSLP antigens can be derived from various animal species, including humans, primates, mice, rats, rabbits, or other mammals. In some embodiments, the TSLP molecule is human TSLP. In some embodiments, the TSLP molecule is rat TSLP. In some embodiments, the TSLP molecule is cynomolgus macaque TSLP. In some embodiments, the TSLP molecule is wild-type. In some embodiments, the TSLP molecule is a mutant (e.g., comprising one or more insertions, deletions, and/or amino acid substitutions compared to the wild-type TSLP sequence from the same species).

在一些實施例中，該抗TSLP抗體部分以≤ 1 μM，諸如≤100 nM，較佳為≤10 nM，更佳為≤1 nM的Kd結合至TSLP抗原。舉例而言，該抗TSLP抗體部分之Kd值係介於約1 nM與約1 pM之間。在一些實施例中，該抗TSLP抗體部分以約1×10-11M至約1×10-7M (例如，約1×10-11M至約1×10-10M、約1×10-11M至約1×10-9M、約1×10-11M至約1×10-8M、約1×10-10M至約1×10-9M、約1×10-10M至約1×10-8M、約1×10-10M至約1×10-7M、約1×10-9M至約1×10-7M、約1×10-9M至約1×10-8M或約1×10-8M至約1×10-7M)的Kd結合至人類TSLP抗原及/或猴(例如，食蟹獼猴) TSLP抗原。In some embodiments, the anti-TSLP antibody portion binds to the TSLP antigen with aKd of ≤ 1 μM, such as ≤ 100 nM, preferably ≤ 10 nM, and more preferably ≤ 1 nM. For example, the anti-TSLP antibody portion has aKd value between about 1 nM and about 1 pM. In some embodiments, the anti-TSLP antibody portion binds to a human TSLP antigen and/or a monkey (e.g., acynomolgus macaque) TSLP antigen witha Kd of about 1×10-11 M to about 1×10-10M( e.g., about 1×10-11 M to about 1×10-10 M, about 1×10-11 M to about 1×10-9 M, about 1×10-11 M to about 1×10-8 M, about 1×10-10 M to about 1×10-9 M, about 1×10-10 M to about 1×10-8 M, about 1×10-10 M to about 1×10-7 M, about 1×10-9 M to about 1×10-7 M, about 1×10-9M to about 1×10-8 M, or about 1×10-8 M to about 1×10-7 M).

本文所述之抗TSLP抗體部分可為任何形式並衍生自任何合適的抗TSLP抗體或其抗原結合片段。舉例而言，該抗TSLP抗體部分可選自全長抗體、scFv、VH、VL、scFv-scFv、Fv、Fab、Fab’、(Fab’)2、微型抗體(minibody)、雙抗體、域抗體變體(dAb)、單域抗體(sdAb)、駱駝科抗體(VHH)、纖維接合素3域變體、錨蛋白重複變體及其他衍生自其他蛋白質框架的抗原特異性結合域。在一些實施例中，該抗TSLP抗體部分為Fab。在一些實施例中，該抗TSLP抗體部分為scFv。在一些實施例中，該抗TSLP抗體部分係人源化。在一些實施例中，該抗TSLP抗體部分係嵌合。在一些實施例中，該抗TSLP抗體部分係衍生自小鼠、大鼠、猴或兔的單株抗體。在一些實施例中，該抗TSLP抗體部分係衍生自本領域已知之任何抗TSLP 單株抗體。在一些實施例中，該抗TSLP抗體部分係衍生自完整人類抗體，例如使用噬菌體展示、酵母菌展示或帶有人類Ig基因的轉基因小鼠開發。在一些實施例中，該抗TSLP抗體部分為單特異性抗體、多特異性抗體、單價抗體或多價(例如，雙價)抗體。The anti-TSLP antibody portions described herein can be in any form and derived from any suitable anti-TSLP antibody or antigen-binding fragment thereof. For example, the anti-TSLP antibody portion can be selected from a full-length antibody, scFv, VH, VL, scFv-scFv, Fv, Fab, Fab', (Fab')2 , minibody, diabody, domain antibody variant (dAb), single domain antibody (sdAb), camelid antibody (VHH), fibronectin 3-domain variant, anaglycin repeat variant, and other antigen-specific binding domains derived from other protein frameworks. In some embodiments, the anti-TSLP antibody portion is a Fab. In some embodiments, the anti-TSLP antibody portion is a scFv. In some embodiments, the anti-TSLP antibody portion is humanized. In some embodiments, the anti-TSLP antibody portion is chimeric. In some embodiments, the anti-TSLP antibody portion is derived from a mouse, rat, monkey, or rabbit monoclonal antibody. In some embodiments, the anti-TSLP antibody portion is derived from any anti-TSLP monoclonal antibody known in the art. In some embodiments, the anti-TSLP antibody portion is derived from a fully human antibody, e.g., developed using phage display, yeast display, or transgenic mice harboring human Ig genes. In some embodiments, the anti-TSLP antibody portion is a monospecific antibody, a multispecific antibody, a monovalent antibody, or a multivalent (e.g., bivalent) antibody.

在一些實施例中，該抗TSLP抗體部分特異性地結合至人類及非人類靈長類動物(諸如食蟹獼猴) TSLP。在一些實施例中，該抗TSLP抗體部分僅結合至人類TSLP。示例性抗TSLP抗體及其抗原結合片段包括但不限於TEZSPIRE™ (特澤佩魯單抗-ekko (tezepelumab-ekko))及以下揭露的內容：WO2023098491；US20220289833；WO2021104053；US20230029835；US20180327489；US20220340654；US20230201120；US 7,982,016；WO2022166739；US20110305705；US20120190829；US20130023647；WO2022157773；WO2022254428；WO2016142426；WO2007096149；US 9,346,870；US 8,512,705；US 10,828,365；WO2017042701；US20220177565；及US 10,745,473，其各自之內容皆通過引用具體併入本文中。含有與猴TSLP具有交叉反應性的此類抗TSLP抗體部分的多特異性構築體(諸如本文所述之任何多特異性構築體)可有助於非人類靈長類動物的毒性研究，其可為人類臨床試驗候選者提供更相關的安全性評估，而無需在黑猩猩身上進行毒性研究或使用替代分子。在一些實施例中，該抗TSLP抗體部分與人類TSLP的結合比非人類(例如，食蟹獼猴) TSLP的更強(諸如至少約1.5、2、5、10、20、50、100、1000或更多倍之任一者)。In some embodiments, the anti-TSLP antibody portion specifically binds to human and non-human primate (e.g., cynomolgus macaque) TSLP. In some embodiments, the anti-TSLP antibody portion binds only to human TSLP. Exemplary anti-TSLP antibodies and antigen-binding fragments thereof include, but are not limited to, TEZSPIRE™ (tezepelumab-ekko) and the following disclosures: WO2023098491; US20220289833; WO2021104053; US20230029835; US20180327489; US20220340654; US20230201120; US 7,982,016; WO2022166739; US20110305705; US20120190829; US20130023647; WO2022157773; WO2022254428; WO2016142426; WO2007096149; US 9,346,870; US 8,512,705; US 10,828,365; WO2017042701; US20220177565; and US 10,745,473, the contents of each of which are specifically incorporated herein by reference. Multispecific constructs containing such anti-TSLP antibody portions that are cross-reactive with monkey TSLP (such as any of the multispecific constructs described herein) can facilitate toxicity studies in non-human primates, which can provide more relevant safety assessments for human clinical trial candidates without the need for toxicity studies in chimpanzees or the use of surrogate molecules. In some embodiments, the anti-TSLP antibody portion binds to human TSLP more strongly (e.g., at least about 1.5, 2, 5, 10, 20, 50, 100, 1000, or more times) than to non-human (e.g., cynomolgus macaque) TSLP.

抗TSLP抗體部分可完全或部分調節、阻斷、抑制、降低、拮抗、中和，或干擾TSLP的功能活性。在抗TSLP抗體部分存在下，當TSLP的功能活性降低至少約95% (諸如至少約96%、97%、98%、99%或100%之任一者)時，與不被抗TSLP抗體部分結合時相比，該抗TSLP抗體部分被視為能夠完全調控、阻斷、抑制、降低、拮抗、中和或干擾TSLP的功能活性。在抗TSLP抗體部分存在下，當TSLP的功能活性降低至少約50% (諸如至少約55%、60%、70%、75%、80%、85%或90%之任一者)時，與不被抗TSLP抗體部分結合時相比，該抗TSLP抗體部分被視為能夠顯著調控、阻斷、抑制、降低、拮抗、中和或干擾TSLP的功能活性。在抗TSLP抗體部分存在下，當標靶抗原的功能活性降低小於約50% (諸如降低小於約10%、15%、20%、25%、30%、35%、或40%或45%之任一者)時，與不被抗TSLP抗體部分結合時相比，該抗TSLP抗體部分被視為能夠部分調控、阻斷、抑制、降低、拮抗、中和或干擾TSLP的功能活性。The anti-TSLP antibody portion can fully or partially modulate, block, inhibit, reduce, antagonize, neutralize, or interfere with the functional activity of TSLP. In the presence of the anti-TSLP antibody portion, when the functional activity of TSLP is reduced by at least about 95% (e.g., at least about any of 96%, 97%, 98%, 99%, or 100%) compared to when not bound by the anti-TSLP antibody portion, the anti-TSLP antibody portion is considered capable of fully modulating, blocking, inhibiting, reducing, antagonizing, neutralizing, or interfering with the functional activity of TSLP. When the functional activity of TSLP is reduced by at least about 50% (e.g., at least about any of 55%, 60%, 70%, 75%, 80%, 85%, or 90%) in the presence of an anti-TSLP antibody portion compared to when not bound by the anti-TSLP antibody portion, the anti-TSLP antibody portion is considered capable of significantly modulating, blocking, inhibiting, reducing, antagonizing, neutralizing, or interfering with the functional activity of TSLP. When the functional activity of the target antigen is reduced by less than about 50% (e.g., by less than about 10%, 15%, 20%, 25%, 30%, 35%, or 40% or 45%) in the presence of an anti-TSLP antibody portion compared to when not bound by the anti-TSLP antibody portion, the anti-TSLP antibody portion is considered capable of partially modulating, blocking, inhibiting, reducing, antagonizing, neutralizing, or interfering with the functional activity of TSLP.

在一些實施例中，該抗TSLP抗體部分在序列中含有一或多個變異。在一些實施例中，該變體序列中的胺基酸變異(例如，取代)不會實質上降低 (例如，降低至多約50%、40%、30%、20%、10%、5%、1%或更少之任一者)抗TSLP抗體部分結合至TSLP的能力。亦考慮實質上改善(例如，改善至少約1.2、1.5、2、5、10、20、50 或更多倍之任一者)抗TSLP抗體部分與TSLP之結合親和力或其他性質(諸如特異性、免疫原性及/或與TSLP表位變體的交叉反應)的修飾。In some embodiments, the anti-TSLP antibody portion contains one or more variations in sequence. In some embodiments, the amino acid variations (e.g., substitutions) in the variant sequence do not substantially reduce (e.g., by at most about 50%, 40%, 30%, 20%, 10%, 5%, 1%, or less) the ability of the anti-TSLP antibody portion to bind to TSLP. Modifications that substantially improve (e.g., by at least about 1.2, 1.5, 2, 5, 10, 20, 50, or more fold) the binding affinity or other properties of the anti-TSLP antibody portion to TSLP, such as specificity, immunogenicity, and/or cross-reactivity with TSLP epitope variants, are also contemplated.

在一些實施例中，該抗TSLP抗體部分為scFv (本文中亦稱為「抗TSLP scFv」)。在一些實施例中，該抗TSLP scFv之N端至C端包含：VH-視情況的連接子-VL，或VL-視情況的連接子-VH。以下「連接子」小節中的任何連接子皆可用於連接抗TSLP scFv的VH及VL。In some embodiments, the anti-TSLP antibody portion is a scFv (also referred to herein as an "anti-TSLP scFv"). In some embodiments, the anti-TSLP scFv comprises, from the N-terminus to the C-terminus, VH-optional linker-VL, or VL-optional linker-VH. Any linker described below in the "Linker" section can be used to link the VH and VL of the anti-TSLP scFv.

在一些實施例中，該抗TSLP抗體部分為Fab片段(本文中亦稱為「抗TSLP Fab片段」或「抗TSLP Fab」)，其包含含有VH及CH1的第一多肽，以及含有VL及CL的第二多肽。在一些實施例中，該抗TSLP Fab片段內之可變區及恆定區的配置可能與天然抗TSLP Fab片段中發現的不同。舉例而言，在一些實施例中，該抗TSLP Fab片段包含含有VH及CL的第一多肽，以及含有VL及CH1的第二多肽(參見例如Shaefer等人(2011)，PNAS，108:111870-92，其內容通過引用整體併入本文)。In some embodiments, the anti-TSLP antibody portion is a Fab fragment (also referred to herein as an "anti-TSLP Fab fragment" or "anti-TSLP Fab") comprising a first polypeptide comprising VH and CH1, and a second polypeptide comprising VL and CL. In some embodiments, the configuration of the variable and constant regions within the anti-TSLP Fab fragment may differ from that found in native anti-TSLP Fab fragments. For example, in some embodiments, the anti-TSLP Fab fragment comprises a first polypeptide comprising VH and CL, and a second polypeptide comprising VL and CH1 (see, e.g., Shaefer et al. (2011), PNAS, 108:111870-92, the contents of which are incorporated herein by reference in their entirety).

在一些實施例中，該抗TSLP中的CH1及VH與VL及CL進行異二聚化，並藉由重鏈與輕鏈恆定區之間的雙硫鍵共價連接。在一些實施例中，該抗TSLP Fab片段具有基本結構NH2-VL-CL-S-S-CH1-VH-NH2。在一些實施例中，該抗TSLP Fab的CH1及CL藉由一或多個雙硫鍵連接。在一些實施例中，該抗TSLP Fab片段之CH1與CL之間的雙硫鍵數量為至少1個，諸如2、3、4、5個或更多。在一些實施例中，該抗TSLP Fab片段(諸如CH1及CL區域)中的半胱胺酸殘基係經工程改造以導入雙硫鍵。In some embodiments, the CH1 and VH of the anti-TSLP Fab heterodimerize with the VL and CL and are covalently linked by disulfide bonds between the heavy and light chain constant regions. In some embodiments, the anti-TSLP Fab fragment has the basic structureNH2 -VL-CL-SS-CH1-VH-NH2 . In some embodiments, the CH1 and CL of the anti-TSLP Fab are linked by one or more disulfide bonds. In some embodiments, the number of disulfide bonds between the CH1 and CL of the anti-TSLP Fab fragment is at least one, such as two, three, four, five, or more. In some embodiments, cysteine residues in the anti-TSLP Fab fragment (such as the CH1 and CL regions) are engineered to introduce disulfide bonds.

在一些實施例中，該抗TSLP Fab片段在C端處不含雙硫鍵。舉例而言，該抗TSLP Fab片段之重鏈及輕鏈可以此方式進行工程改造，以便穩定相互作用而不需要雙硫鍵。舉例而言，重鏈及輕鏈可以此方式進行工程改造，以便穩定相互作用而不需要雙硫鍵。在一些實施例中，重鏈或輕鏈可經工程改造以移除半胱胺酸殘基，且其中重鏈及輕鏈仍然穩定地相互作用並發揮Fab的功用。在一些實施例中，進行突變以促進重鏈與輕鏈之間的穩定相互作用。舉例而言，「杵臼」工程改造策略可用於促進Fab的重鏈與輕鏈之間的二聚化(參見例如1996 Protein Engineering，9:617 - 621)。本文亦考慮使用針對特定目的而設計的變體Fab片段，例如CH1及/或CL之恆定域的胺基酸變化，以及移除雙硫鍵或添加用於純化的標籤等。在以下「恆定域及Fc域」小節中更詳細地描述此等突變。In some embodiments, the anti-TSLP Fab fragment does not contain a disulfide bond at the C-terminus. For example, the heavy and light chains of the anti-TSLP Fab fragment can be engineered in such a way that they interact stably without a disulfide bond. For example, the heavy and light chains can be engineered in such a way that they interact stably without a disulfide bond. In some embodiments, the heavy or light chain can be engineered to remove a cysteine residue, and the heavy and light chains still interact stably and function as a Fab. In some embodiments, mutations are made to promote a stable interaction between the heavy and light chains. For example, a "knob-in-hole" engineering strategy can be used to promote dimerization between the heavy and light chains of a Fab (see, e.g., Protein Engineering, 1996, 9:617-621). This article also contemplates the use of variant Fab fragments designed for specific purposes, such as amino acid changes in the homeostasis domains of CH1 and/or CL, removal of disulfide bonds, or addition of tags for purification. These mutations are described in more detail in the "Homeostasis Domain and Fc Domain" section below.

在一些實施例中，該抗TSLP Fab片段之CH1及CL由約2個雙硫鍵連接。在一些實施例中，該抗TSLP Fab片段包含人類免疫球蛋白CH1，例如含有SEQ ID NO:158-162之任一者之胺基酸序列。在一些實施例中，該抗TSLP Fab片段包含人類λ輕鏈恆定區，例如含有SEQ ID NO:74或75之胺基酸序列。在一些實施例中，該抗TSLP Fab片段包含人類κ輕鏈恆定區，例如含有SEQ ID NO:73之胺基酸序列。In some embodiments, the CH1 and CL of the anti-TSLP Fab fragment are linked by about two disulfide bonds. In some embodiments, the anti-TSLP Fab fragment comprises a human immunoglobulin CH1, e.g., comprising the amino acid sequence of any one of SEQ ID NOs: 158-162. In some embodiments, the anti-TSLP Fab fragment comprises a human lambda light chain constant region, e.g., comprising the amino acid sequence of SEQ ID NOs: 74 or 75. In some embodiments, the anti-TSLP Fab fragment comprises a human kappa light chain constant region, e.g., comprising the amino acid sequence of SEQ ID NO: 73.

在一些實施例中，該抗TSLP抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，其中該VH包含(i)含有SEQ ID NO:2之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:3之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；且該VL包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該VH包含(i)含有SEQ ID NO:2之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:3之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3。在一些實施例中，該抗TSLP抗體部分包含(a)含有與SEQ ID NO:1具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:5具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；或(b)含有與SEQ ID NO:9具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:10具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:1之胺基酸序列的VH及含有SEQ ID NO:5之胺基酸序列的VL；或(b)含有SEQ ID NO:9之胺基酸序列的VH及含有SEQ ID NO:10之胺基酸序列的VL。In some embodiments, the anti-TSLP antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 2, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 3, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 4. NO:4 or a variant thereof containing up to about 3 (e.g., 1, 2 or 3) amino acid variations (e.g., insertions, deletions and/or substitutions, such as conservative substitutions); and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:6 or a variant thereof containing up to about 3 (e.g., 1, 2 or 3) amino acid variations (e.g., insertions, deletions and/or substitutions, such as conservative substitutions); (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:7 or a variant thereof containing up to about 3 (e.g., 1, 2 or 3) amino acid variations (e.g., insertions, deletions and/or substitutions, such as conservative substitutions); and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:8 or a variant thereof containing up to about 3 (e.g., 1, 2 or 3) amino acid variations (e.g., insertions, deletions and/or substitutions, such as conservative substitutions); NO:8 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:2; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:3; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:4; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:6; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:7; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:8. In some embodiments, the anti-TSLP antibody portion comprises (a) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 1 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 5; or (b) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 9 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 10. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH comprising the amino acid sequence of SEQ ID NO: 1 and a VL comprising the amino acid sequence of SEQ ID NO: 5; or (b) a VH comprising the amino acid sequence of SEQ ID NO: 9 and a VL comprising the amino acid sequence of SEQ ID NO: 10.

在一些實施例中，該抗TSLP抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，其中該VH包含(i)含有SEQ ID NO:12之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:13之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；且該VL包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該VH包含(i)含有SEQ ID NO:12之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:13之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3。在一些實施例中，該抗TSLP抗體部分包含含有與SEQ ID NO:11具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:15具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:11之胺基酸序列的VH及含有SEQ ID NO:15之胺基酸序列的VL。In some embodiments, the anti-TSLP antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 12, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 13, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 14. NO:4 or a variant thereof containing up to about 3 (e.g., 1, 2 or 3) amino acid variations (e.g., insertions, deletions and/or substitutions, such as conservative substitutions); and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:6 or a variant thereof containing up to about 3 (e.g., 1, 2 or 3) amino acid variations (e.g., insertions, deletions and/or substitutions, such as conservative substitutions); (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:7 or a variant thereof containing up to about 3 (e.g., 1, 2 or 3) amino acid variations (e.g., insertions, deletions and/or substitutions, such as conservative substitutions); and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:8 or a variant thereof containing up to about 3 (e.g., 1, 2 or 3) amino acid variations (e.g., insertions, deletions and/or substitutions, such as conservative substitutions); NO: 8, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 12; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 13; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 4; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 6; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 7; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 8. In some embodiments, the anti-TSLP antibody portion comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 11 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 15. In some embodiments, the anti-TSLP antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 11 and a VL comprising the amino acid sequence of SEQ ID NO: 15.

在一些實施例中，該抗TSLP抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，其中該VH包含(i)含有SEQ ID NO:28之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:29之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:30之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；且該VL包含(i)含有SEQ ID NO:32之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:33之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:34之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該VH包含(i)含有SEQ ID NO:28之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:29之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:30之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:32之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:33之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:34之胺基酸序列的CDR-L3。在一些實施例中，該抗TSLP抗體部分包含含有與SEQ ID NO:27具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:31具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:27之胺基酸序列的VH及含有SEQ ID NO:31之胺基酸序列的VL。In some embodiments, the anti-TSLP antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 28, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 29, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 21. NO: 30 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 32 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 33 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 34 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); In some embodiments, the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 28; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 29; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 30; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 32; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 33; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 34. In some embodiments, the anti-TSLP antibody portion comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 27 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 31. In some embodiments, the anti-TSLP antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 27 and a VL comprising the amino acid sequence of SEQ ID NO: 31.

在一些實施例中，該抗TSLP抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，其中該VH包含(i)含有SEQ ID NO:36之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:37之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:38之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；且該VL包含(i)含有SEQ ID NO:40之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:41之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:42之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該VH包含(i)含有SEQ ID NO:36之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:37之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:38之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:40之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:41之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:42之胺基酸序列的CDR-L3。在一些實施例中，該抗TSLP抗體部分包含含有與SEQ ID NO:35具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:39具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:35之胺基酸序列的VH及含有SEQ ID NO:39之胺基酸序列的VL。In some embodiments, the anti-TSLP antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 36, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 37, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 38. NO: 38 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 40 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 41 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 42 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); In some embodiments, the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 36; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 37; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 38; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 40; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 41; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 42. In some embodiments, the anti-TSLP antibody portion comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about any of 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 35 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about any of 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 39. In some embodiments, the anti-TSLP antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 35 and a VL comprising the amino acid sequence of SEQ ID NO: 39.

在一些實施例中，該抗TSLP抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，其中該VH包含(i)含有SEQ ID NO:44之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:45之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:46之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；且該VL包含(i)含有SEQ ID NO:48之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:49之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:50之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該VH包含(i)含有SEQ ID NO:44之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:45之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:46之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:48之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:49之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:50之胺基酸序列的CDR-L3。在一些實施例中，該抗TSLP抗體部分包含含有與SEQ ID NO:43具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:47具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL。在一些實施例中，該抗TSLP抗體部分包含含有SEQ ID NO:43之胺基酸序列的VH及含有SEQ ID NO:47之胺基酸序列的VL。In some embodiments, the anti-TSLP antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 44, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 45, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 46. NO: 46 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 48 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 49 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 49 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); NO: 50 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 44; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 45; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 46; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 48; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 49; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 50. In some embodiments, the anti-TSLP antibody portion comprises a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 43 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to SEQ ID NO: 47. In some embodiments, the anti-TSLP antibody portion comprises a VH comprising the amino acid sequence of SEQ ID NO: 43 and a VL comprising the amino acid sequence of SEQ ID NO: 47.

在一些實施例中，該抗TSLP抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，其中該VH包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；且該VL包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3或其含有至多約3個(例如，1、2或3個)胺基酸變異(例如，插入、缺失及/或取代，諸如保留性取代)的變體。在一些實施例中，該VH包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3。在一些實施例中，該抗TSLP抗體部分包含(a)含有與SEQ ID NO:19具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:23具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(b)含有與SEQ ID NO:63具有至少約80%序列同一性之胺基酸序列的VH及含有與SEQ ID NO:53具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(c)含有與SEQ ID NO:63具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:54具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(d)含有與SEQ ID NO:56具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:52具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(e)含有與SEQ ID NO:55具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:51具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(f)含有與SEQ ID NO:56具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:51具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(g)含有與SEQ ID NO:63具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:51具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(h)含有與SEQ ID NO:64具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:51具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(i)含有與SEQ ID NO:55具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:52具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(j)含有與SEQ ID NO:58具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:52具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(k)含有與SEQ ID NO:60具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:52具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(l)含有與SEQ ID NO:64具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:52具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(m)含有與SEQ ID NO:55具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:53具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(n)含有與SEQ ID NO:57具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:53具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(o)含有與SEQ ID NO:58具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:53具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(p)含有與SEQ ID NO:62具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:53具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(q)含有與SEQ ID NO:64具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:53具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(r)含有與SEQ ID NO:55具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:54具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(s)含有與SEQ ID NO:61具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:54具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(t)含有與SEQ ID NO:62具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:54具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(u)含有與SEQ ID NO:64具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:54具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(v)含有與SEQ ID NO:63具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:52具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(w)含有與SEQ ID NO:188具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:189具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；(x)含有與SEQ ID NO:188具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:190具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL；或(xi)含有與SEQ ID NO:228具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VH及含有與SEQ ID NO:190具有至少約80% (諸如至少約85%、90%、95%、96%、97%、98%、99%或更多之任一者)序列同一性之胺基酸序列的VL。在一些實施例中，該抗TSLP抗體部分包含：(a)含有SEQ ID NO:19之胺基酸序列的VH及含有SEQ ID NO:23之胺基酸序列的VL；(b)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(c)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(d)含有SEQ ID NO:56之胺基酸序列的VH及含有SEQ ID NO:52之胺基酸序列的VL；(e)含有SEQ ID NO:55之胺基酸序列的VH及含有SEQ ID NO:51之胺基酸序列的VL；(f)含有SEQ ID NO:56之胺基酸序列的VH及含有SEQ ID NO:51之胺基酸序列的VL；(g)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:51之胺基酸序列的VL；(h)含有SEQ ID NO:64之胺基酸序列的VH及含有SEQ ID NO:51之胺基酸序列的VL；(i)含有SEQ ID NO:55之胺基酸序列的VH及含有SEQ ID NO:52之胺基酸序列的VL；(j)含有SEQ ID NO:58之胺基酸序列的VH及含有SEQ ID NO:52之胺基酸序列的VL；(k)含有SEQ ID NO:60之胺基酸序列的VH及含有SEQ ID NO:52之胺基酸序列的VL；(l)含有SEQ ID NO:64之胺基酸序列的VH及含有SEQ ID NO:52之胺基酸序列的VL；(m)含有SEQ ID NO:55之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(n)含有SEQ ID NO:57之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(o)含有SEQ ID NO:58之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(p)含有SEQ ID NO:62之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(q)含有SEQ ID NO:64之胺基酸序列的VH及含有SEQ ID NO:53之胺基酸序列的VL；(r)含有SEQ ID NO:55之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(s)含有SEQ ID NO:61之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(t)含有SEQ ID NO:62之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(u)含有SEQ ID NO:64之胺基酸序列的VH及含有SEQ ID NO:54之胺基酸序列的VL；(v)含有SEQ ID NO:63之胺基酸序列的VH及含有SEQ ID NO:52之胺基酸序列的VL；(w)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:189之胺基酸序列的VL；(x)含有SEQ ID NO:188之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL；或(xi)含有SEQ ID NO:228之胺基酸序列的VH及含有SEQ ID NO:190之胺基酸序列的VL。在一些實施例中，該VH進一步包含位於位置44處的取代及/或該VL進一步包含位於位置100處的取代。在一些實施例中，該VH進一步包含位於位置G44或R44處的取代及/或該VL進一步包含位於位置Q100處的取代。在一些實施例中，該取代為G44C或R44C及/或Q100C。In some embodiments, the anti-TSLP antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21, or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22. NO: 22 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); and (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 26 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions); NO: 26 or a variant thereof containing up to about 3 (e.g., 1, 2, or 3) amino acid variations (e.g., insertions, deletions, and/or substitutions, such as conservative substitutions). In some embodiments, the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 22; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 25; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 26. In some embodiments, the anti-TSLP antibody portion comprises (a) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 19 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 23; (b) a VH comprising an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 63 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 53; (c) a VH comprising an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 63 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: (e) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 54; (f) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 56; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 52; (g) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 55; (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 51; (f) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 56; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 51; (g) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 63; (i) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 51 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 51; (ii) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 51; (i) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 52; (ii) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 58; and (iii) a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 52; (iv) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 58; and (v) a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 52; (i) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64; (ii) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 52; (iii) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64; (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 53; (n) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 57; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 53; (o) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 58; (a) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 53; (b) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 62; and (c) a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 53; (d) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64; (i) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 53; (ii) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 55; and (iii) a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 54; and (iv) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 55; (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 54; (t) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 62; and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 54; (u) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64; (v) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 63 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64; (v) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 63 and a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64; (v) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 64; (i) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 189; (ii) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 188; and (iii) a VL comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 190; or (iv) a VH comprising an amino acid sequence having at least about 80% (e.g., at least about 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) sequence identity to SEQ ID NO: 191. NO:228 has a VH having an amino acid sequence that is at least about 80% (e.g., at least about any of 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO:190 and a VL comprising an amino acid sequence that is at least about 80% (e.g., at least about any of 85%, 90%, 95%, 96%, 97%, 98%, 99% or more) identical to SEQ ID NO:190. In some embodiments, the anti-TSLP antibody portion comprises: (a) a VH comprising the amino acid sequence of SEQ ID NO: 19 and a VL comprising the amino acid sequence of SEQ ID NO: 23; (b) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 53; (c) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 54; (d) a VH comprising the amino acid sequence of SEQ ID NO: 56 and a VL comprising the amino acid sequence of SEQ ID NO: 52; (e) a VH comprising the amino acid sequence of SEQ ID NO: 55 and a VL comprising the amino acid sequence of SEQ ID NO: 51; (f) a VH comprising the amino acid sequence of SEQ ID NO: 56 and a VL comprising the amino acid sequence of SEQ ID NO: 51; (g) a VH comprising the amino acid sequence of SEQ ID NO: 56 and a VL comprising the amino acid sequence of SEQ ID NO: NO:63 and a VL comprising the amino acid sequence of SEQ ID NO:51; (h) a VH comprising the amino acid sequence of SEQ ID NO:64 and a VL comprising the amino acid sequence of SEQ ID NO:51; (i) a VH comprising the amino acid sequence of SEQ ID NO:55 and a VL comprising the amino acid sequence of SEQ ID NO:52; (j) a VH comprising the amino acid sequence of SEQ ID NO:58 and a VL comprising the amino acid sequence of SEQ ID NO:52; (k) a VH comprising the amino acid sequence of SEQ ID NO:60 and a VL comprising the amino acid sequence of SEQ ID NO:52; (l) a VH comprising the amino acid sequence of SEQ ID NO:64 and a VL comprising the amino acid sequence of SEQ ID NO:52; (m) a VH comprising the amino acid sequence of SEQ ID NO:55 and a VL comprising the amino acid sequence of SEQ ID NO:53; (n) a VH comprising the amino acid sequence of SEQ ID NO:56 and a VL comprising the amino acid sequence of SEQ ID NO:57; NO:57 and a VL comprising the amino acid sequence of SEQ ID NO:53; (o) a VH comprising the amino acid sequence of SEQ ID NO:58 and a VL comprising the amino acid sequence of SEQ ID NO:53; (p) a VH comprising the amino acid sequence of SEQ ID NO:62 and a VL comprising the amino acid sequence of SEQ ID NO:53; (q) a VH comprising the amino acid sequence of SEQ ID NO:64 and a VL comprising the amino acid sequence of SEQ ID NO:53; (r) a VH comprising the amino acid sequence of SEQ ID NO:55 and a VL comprising the amino acid sequence of SEQ ID NO:54; (s) a VH comprising the amino acid sequence of SEQ ID NO:61 and a VL comprising the amino acid sequence of SEQ ID NO:54; (t) a VH comprising the amino acid sequence of SEQ ID NO:62 and a VL comprising the amino acid sequence of SEQ ID NO:54; (u) a VH comprising the amino acid sequence of SEQ ID NO:64 (i) a VH comprising the amino acid sequence of SEQ ID NO: 64 and a VL comprising the amino acid sequence of SEQ ID NO: 54; (ii) a VH comprising the amino acid sequence of SEQ ID NO: 63 and a VL comprising the amino acid sequence of SEQ ID NO: 52; (iii) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 189; (iv) a VH comprising the amino acid sequence of SEQ ID NO: 188 and a VL comprising the amino acid sequence of SEQ ID NO: 190; or (v) a VH comprising the amino acid sequence of SEQ ID NO: 228 and a VL comprising the amino acid sequence of SEQ ID NO: 190. In some embodiments, the VH further comprises a substitution at position 44 and/or the VL further comprises a substitution at position 100. In some embodiments, the VH further comprises a substitution at position G44 or R44 and/or the VL further comprises a substitution at position Q100. In some embodiments, the substitution is G44C or R44C and/or Q100C.

在一些實施例中，該抗TSLP抗體部分為全長抗體(「抗TSLP全長抗體」)。在一些實施例中，該抗TSLP全長抗體包含人類κ CL或人類λ CL，諸如含有SEQ ID NO:73-75之任一者之胺基酸序列的CL。在一些實施例中，該抗TSLP全長抗體包含衍生自人類IgG1、IgG2、IgG3或IgG4 (諸如人類IgG1)的Fc域。以下「Fc域」小節中所述之任何Fc域皆可在此使用。在一些實施例中，該Fc域包含：i) L234A+L235A (「LALA」)突變，ii) M252Y+S254T+T256E (「YTE」)突變，及/或iii) M428L+N434S (「LS」)突變，其係根據EU編號。在一些實施例中，該Fc域進一步包含杵臼突變，諸如在一個Fc次單元中的T366W 「杵」突變，以及在另一個Fc次單元中的T366S+L368A+Y407V 「臼」突變。在一些實施例中，i)該Fc域之第一次單元及第二次單元各自含有SEQ ID NO:76-79之任一者的胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；ii)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；或iii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段。在一些實施例中，該第一次單元及該Fc域之第二次單元各自含有SEQ ID NO: 77或79之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段。在一些實施例中，該抗TSLP全長抗體包含：i)兩條各自含有SEQ ID NO:104之胺基酸序列的重鏈或其與SEQ ID NO:104之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈或其與SEQ ID NO:103之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體；ii)兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈或其與SEQ ID NO:105之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈或其與SEQ ID NO:103之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體；或iii)含有SEQ ID NO:136之胺基酸序列的第一重鏈或其與SEQ ID NO:136之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體、含有SEQ ID NO:137之胺基酸序列的第二重鏈或其與SEQ ID NO:137之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈或其與SEQ ID NO:103之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP全長抗體包含：i)兩條含有SEQ ID NO:104之胺基酸序列的重鏈及兩條含有SEQ ID NO:103之胺基酸序列的輕鏈；ii)兩條含有SEQ ID NO:105之胺基酸序列的重鏈及兩條含有SEQ ID NO:103之胺基酸序列的輕鏈；或iii)含有SEQ ID NO:136之胺基酸序列的第一重鏈、含有SEQ ID NO:137之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈。In some embodiments, the anti-TSLP antibody portion is a full-length antibody ("anti-TSLP full-length antibody"). In some embodiments, the anti-TSLP full-length antibody comprises human kappa CL or human lambda CL, such as a CL comprising the amino acid sequence of any one of SEQ ID NOs: 73-75. In some embodiments, the anti-TSLP full-length antibody comprises an Fc domain derived from human IgG1, IgG2, IgG3, or IgG4 (such as human IgG1). Any of the Fc domains described below in the "Fc Domain" section can be used herein. In some embodiments, the Fc domain comprises: i) L234A+L235A ("LALA") mutations, ii) M252Y+S254T+T256E ("YTE") mutations, and/or iii) M428L+N434S ("LS") mutations, according to EU numbering. In some embodiments, the Fc domain further comprises a knob-to-hole mutation, such as a T366W "knob" mutation in one Fc subunit and a T366S+L368A+Y407V "hole" mutation in the other Fc subunit. In some embodiments, i) the first and second Fc domain units each contain the amino acid sequence of any one of SEQ ID NOs: 76-79 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; ii) the first Fc domain unit contains the amino acid sequence of SEQ ID NO: 80 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215, and the second Fc domain unit contains the amino acid sequence of SEQ ID NO: 81 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; or iii) the first Fc domain unit contains the amino acid sequence of SEQ ID NO: 81 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215, and the second Fc domain unit contains the amino acid sequence of SEQ ID NO: 80 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215. In some embodiments, the first unit and the second unit of the Fc domain each contain the amino acid sequence of SEQ ID NO: 77 or 79 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215. In some embodiments, the anti-TSLP full-length antibody comprises: i) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 104, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99% or more) sequence identity to the amino acid sequence of SEQ ID NO: 104, and two light chains each comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99% or more) sequence identity to the amino acid sequence of SEQ ID NO: 103; ii) two heavy chains each comprising the amino acid sequence of SEQ ID NO: 105, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99% or more) sequence identity to the amino acid sequence of SEQ ID NO: 105, and two light chains each comprising the amino acid sequence of SEQ ID NO: 103. NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99% or more) sequence identity to the amino acid sequence of SEQ ID NO: 103; or iii) a first heavy chain comprising the amino acid sequence of SEQ ID NO: 136, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99% or more) sequence identity to the amino acid sequence of SEQ ID NO: 136, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 137, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99% or more) sequence identity to the amino acid sequence of SEQ ID NO: 137, and two light chains each comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99% or more) sequence identity to the amino acid sequence of SEQ ID NO: 103. In some embodiments, the anti-TSLP full-length antibody comprises: i) two heavy chains comprising the amino acid sequence of SEQ ID NO: 104 and two light chains comprising the amino acid sequence of SEQ ID NO: 103; ii) two heavy chains comprising the amino acid sequence of SEQ ID NO: 105 and two light chains comprising the amino acid sequence of SEQ ID NO: 103; or iii) a first heavy chain comprising the amino acid sequence of SEQ ID NO: 136, a second heavy chain comprising the amino acid sequence of SEQ ID NO: 137, and two light chains each comprising the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，該抗TSLP抗體部分為scFv (「抗TSLP scFv」)。在一些實施例中，該抗TSLP scFv包含經由視情況的連接子彼此融合的VH及VL。在一些實施例中，該視情況的連接子包含選自由以下所組成群組之胺基酸序列：GG及SEQ ID NO:14、16-18、97-99、163-172及224之任一者，諸如GG及SEQ ID NO:98、99及224之任一者。在一些實施例中，該抗TSLP抗體部分包含：進一步含有位於位置44處之取代的VH；及/或進一步含有位於位置100處之取代的VL。在一些實施例中，該VH進一步包含位於位置處的取代G44或R44及/或該VL進一步包含位於位置處的取代Q100。在一些實施例中，該取代為G44C或R44C及/或Q100C。在一些實施例中，該抗TSLP scFv含有SEQ ID NO:106、107、218、219、226、227及229之任一者之胺基酸序列或其與SEQ ID NO:106、107、218、219、226、227及229之任一者之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP scFv含有SEQ ID NO:106、107、218、219、226、227及229之任一者之胺基酸序列。In some embodiments, the anti-TSLP antibody portion is a scFv ("anti-TSLP scFv"). In some embodiments, the anti-TSLP scFv comprises a VH and a VL fused to each other via an optional linker. In some embodiments, the optional linker comprises an amino acid sequence selected from the group consisting of GG and any one of SEQ ID NOs: 14, 16-18, 97-99, 163-172, and 224, such as GG and any one of SEQ ID NOs: 98, 99, and 224. In some embodiments, the anti-TSLP antibody portion comprises: a VH further comprising a substitution at position 44; and/or a VL further comprising a substitution at position 100. In some embodiments, the VH further comprises a substitution at position G44 or R44 and/or the VL further comprises a substitution at position Q100. In some embodiments, the substitution is G44C or R44C and/or Q100C. In some embodiments, the anti-TSLP scFv comprises the amino acid sequence of any one of SEQ ID NOs: 106, 107, 218, 219, 226, 227, and 229, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99%, or more) sequence identity to the amino acid sequence of any one of SEQ ID NOs: 106, 107, 218, 219, 226, 227, and 229. In some embodiments, the anti-TSLP scFv contains the amino acid sequence of any one of SEQ ID NOs: 106, 107, 218, 219, 226, 227, and 229.

在一些實施例中，該抗TSLP抗體部分為Fab片段 (「抗TSLP Fab」)。在一些實施例中，該抗TSLP Fab片段包含含有SEQ ID NO:109之胺基酸序列的第一多肽或其與SEQ ID NO:109之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體；及含有SEQ ID NO:103之胺基酸序列的第二多肽或其與SEQ ID NO:103之胺基酸序列具有至少約80% (諸如至少約85%、90%、95%、99%或更多之任一者)序列同一性的變體。在一些實施例中，該抗TSLP Fab片段包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。抗IL-17A抗體部分In some embodiments, the anti-TSLP antibody portion is a Fab fragment ("anti-TSLP Fab"). In some embodiments, the anti-TSLP Fab fragment comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 109; and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103, or a variant thereof having at least about 80% (e.g., at least about 85%, 90%, 95%, 99%, or more) sequence identity to the amino acid sequence of SEQ ID NO: 103. In some embodiments, the anti-TSLP Fab fragment comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103.Anti-IL-17AAntibody Portion

本文所述之抗IL-17A抗體部分之任一者可用作本文所述之多特異性構築體之任一者的第二、第三或第四抗體部分。Any of the anti-IL-17A antibody portions described herein can be used as the second, third, or fourth antibody portion of any of the multispecific constructs described herein.

IL-17A (亦稱為IL-17)為由T輔助17型(Th17)細胞產生的細胞激素，但其亦可由(例如) CD8+ T細胞、自然殺手T細胞、類淋巴組織誘導細胞、先天類淋巴細胞、γδ T細胞及上皮細胞產生。IL-17A涉及Th17發炎反應(諸如宿主防禦及自體免疫功能)，並已被確定為治療多種自體免疫病症的可能治療標靶，例如類風濕性關節炎(rheumatoid arthritis)、多發性硬化症(multiple sclerosis)、發炎性腸道疾病(inflammatory bowel disease，IBD)、乾癬(psoriasis)、乾癬性關節炎(psoriatic arthritis)及僵直性脊柱炎(ankylosing spondylitis)。此示例性抗原可用作特定發炎性疾病(例如，過敏或自體免疫疾病)發展的生物標記、預後指標及抗原表現發炎性疾病的免疫治療標靶。IL-17A (also known as IL-17) is a cytokine produced by T helper type 17 (Th17) cells, but it can also be produced by, for example, CD8+ T cells, natural killer T cells, lymphoid tissue-inducing cells, innate lymphoid cells, γδ T cells, and epithelial cells. IL-17A is involved in Th17 inflammatory responses (e.g., host defense and autoimmune functions) and has been identified as a potential therapeutic target for treating a variety of autoimmune disorders, such as rheumatoid arthritis, multiple sclerosis, inflammatory bowel disease (IBD), psoriasis, psoriasis arthritis, and ankylosing spondylitis. This exemplary antigen can be used as a biomarker for the development of specific inflammatory diseases (e.g., allergies or autoimmune diseases), a prognostic indicator, and as a target for immunotherapy of antigen-expressing inflammatory diseases.

在一些實施例中，該抗IL-17A抗體部分特異性地結合至人類及非人類靈長類動物(諸如食蟹獼猴) IL-17A。在一些實施例中，該抗IL-17A抗體部分僅結合至人類IL-17A。示例性抗IL-17A抗體及其抗原結合片段包括但不限於COSENTYX® (蘇金單抗(secukinumab))、Taltz® (依奇珠單抗(ixekizumab))、Bimzelx® (比美吉珠單抗(bimekizumab))及以下揭露的內容：WO2019154420；WO2013150043；WO2014161570A1；WO2023035272A1；WO2006013107；WO2014144280；WO2009082624；WO2008047134；WO2009130459；US 8,865,166；US 9,481,736；US 9,193,788；及US 9,655,964，其各自之內容皆通過引用具體併入本文中。抗IL-17F抗體部分In some embodiments, the anti-IL-17A antibody portion specifically binds to human and non-human primate (e.g., cynomolgus macaque) IL-17A. In some embodiments, the anti-IL-17A antibody portion binds only to human IL-17A. Exemplary anti-IL-17A antibodies and antigen-binding fragments thereof include, but are not limited to, COSENTYX® (secukinumab), Taltz® (ixekizumab), Bimzelx® (bimekizumab), and those disclosed in WO2019154420; WO2013150043; WO2014161570A1; WO2023035272A1; WO2006013107; WO2014144280; WO2009082624; WO2008047134; WO2009130459; US Pat. No. 8,865,166; US Pat. No. 9,481,736; US Pat. No. 9,193,788; and US Pat. No. 116,906,694. 9,655,964, the contents of each of which are specifically incorporated herein by reference.Anti-IL-17FAntibody Portion

本文所述之抗IL-17F抗體部分之任一者可用作本文所述之多特異性構築體之任一者的第二、第三或第四抗體部分。Any of the anti-IL-17F antibody portions described herein can be used as the second, third, or fourth antibody portion of any of the multispecific constructs described herein.

IL-17F為由T輔助17型(Th17)細胞產生的細胞激素，但其亦可由(例如) CD8+ T細胞、自然殺手T細胞、類淋巴組織誘導細胞、先天類淋巴細胞、γδ T細胞及上皮細胞產生。IL-17F常與IL-17A共同表現，並經由與IL-17A相同的受體傳訊(亦即，IL-17RA及IL-17RC)。IL-17F的標靶細胞為上皮細胞、纖維母細胞、角質細胞、滑液膜細胞(synoviocyte)及內皮細胞。IL-17F涉及發炎反應，例如媒介宿主防禦及自體免疫功能，並已被確定為治療氣喘或結腸炎的可能治療標靶。此示例性抗原可用作特定發炎性疾病(例如，過敏或自體免疫疾病)發展的生物標記、預後指標及抗原表現發炎性疾病的免疫治療標靶。IL-17F is a cytokine produced by T helper type 17 (Th17) cells, but it can also be produced by, for example, CD8+ T cells, natural killer T cells, lymphoid tissue-inducing cells, innate lymphoid cells, γδ T cells, and epithelial cells. IL-17F is often co-expressed with IL-17A and signals through the same receptors as IL-17A (i.e., IL-17RA and IL-17RC). IL-17F's target cells include epithelial cells, fibroblasts, keratinocytes, synovial cells (synoviocytes), and endothelial cells. IL-17F is involved in inflammatory responses, such as mediating host defense and autoimmune function, and has been identified as a potential therapeutic target for treating asthma or colitis. This exemplary antigen can be used as a biomarker for the development of specific inflammatory diseases (e.g., allergies or autoimmune diseases), a prognostic indicator, and as a target for immunotherapy of antigen-expressing inflammatory diseases.

在一些實施例中，該抗IL-17F抗體部分特異性地結合至人類及非人類靈長類動物(諸如食蟹獼猴) IL-17F。在一些實施例中，該抗IL-17F抗體部分僅結合至人類IL-17F。示例性抗IL-17F抗體及其抗原結合片段包括但不限於Bimzelx® (比美吉珠單抗)及以下揭露的內容：US 10,829,552；WO2023035272A1；US 9,481,736；WO2009082624；WO2009130459；WO2008047134；及US 9,475,873，其各自之內容皆通過引用具體併入本文中。抗TNF-α抗體部分In some embodiments, the anti-IL-17F antibody portion specifically binds to human and non-human primate (e.g., cynomolgus macaque) IL-17F. In some embodiments, the anti-IL-17F antibody portion binds only to human IL-17F. Exemplary anti-IL-17F antibodies and antigen-binding fragments thereof include, but are not limited to, Bimzelx® (bimelezumab) and the following disclosures: US 10,829,552; WO2023035272A1; US 9,481,736; WO2009082624; WO2009130459; WO2008047134; and US 9,475,873, the contents of each of which are specifically incorporated herein by reference.Anti-TNF-αAntibody Portion

本文所述之抗TNF-α抗體部分之任一者可用作本文所述之多特異性構築體之任一者的第二、第三或第四抗體部分。Any of the anti-TNF-α antibody portions described herein can be used as the second, third, or fourth antibody portion of any of the multispecific constructs described herein.

TNF-α為由活化的巨噬細胞、T淋巴球及自然殺手細胞產生的多效性促發炎性細胞激素，但其亦可由(例如) B細胞、肌肉細胞、纖維母細胞、角質細胞、蝕骨細胞(osteoclast)、心臟細胞、內皮細胞等產生。TNF-α涉及細胞增生、分化、凋亡、調控免疫反應及誘導發炎反應。TNF-α水平的升高與多種自體免疫及發炎性疾病相關，例如乾癬及發炎性腸道疾病(IBD)。此示例性抗原可用作特定發炎性疾病(例如，過敏或自體免疫疾病)發展的生物標記、預後指標及抗原表現發炎性疾病的免疫治療標靶。TNF-α is a pleiotropic pro-inflammatory cytokine produced by activated macrophages, T lymphocytes, and natural killer cells, but it can also be produced by, for example, B cells, muscle cells, fibroblasts, keratinocytes, osteoclasts, cardiac cells, endothelial cells, and the like. TNF-α is involved in cell proliferation, differentiation, apoptosis, regulation of immune responses, and induction of inflammatory responses. Elevated TNF-α levels are associated with a variety of autoimmune and inflammatory diseases, such as pityriasis and inflammatory bowel disease (IBD). This exemplary antigen can be used as a biomarker for the development of specific inflammatory diseases (e.g., allergies or autoimmune diseases), a prognostic indicator, and as a target for immunotherapy of antigen-expressing inflammatory diseases.

在一些實施例中，該抗TNF-α抗體部分特異性地結合至人類及非人類靈長類動物(諸如食蟹獼猴) TNF-α。在一些實施例中，該抗TNF-α抗體部分僅結合至人類TNF-α。示例性抗TNF-α抗體及其抗原結合片段包括但不限於REMICADE™ (英利昔單抗(infliximab))、ENBREL™ (依那西普(etanercept))、HUMIRA™ (阿達木單抗(adalimumab))、CIMZIA® (聚乙二醇化賽妥珠單抗(certolizumab pegol))、SIMPONI® (戈利木單抗(golimumab))及以下揭露的內容：US20190309057；WO2011127141；WO2002012502；US 10,233,238；US 5,656,272；US 6,090,382；US 6,593,458；US 6,498,237；US 6,451,983；US 6,448,380；WO2010121140；US 8,063,182；US 9,655,964；WO1991003553；WO2001094585；US 9,828,424；US 9,481,736；WO2007024705；及WO2012131053，其各自之內容皆通過引用具體併入本文中。抗IFN-γ抗體部分In some embodiments, the anti-TNF-α antibody portion specifically binds to human and non-human primate (e.g., cynomolgus macaque) TNF-α. In some embodiments, the anti-TNF-α antibody portion binds only to human TNF-α. Exemplary anti-TNF-α antibodies and antigen-binding fragments thereof include, but are not limited to, REMICADE™ (infliximab), ENBREL™ (etanercept), HUMIRA™ (adalimumab), CIMZIA® (certolizumab pegol), SIMPONI® (golimumab), and those disclosed in: US20190309057; WO2011127141; WO2002012502; US 10,233,238; US 5,656,272; US 6,090,382; US 6,593,458; US 6,498,237; US 6,451,983; US 6,448,380; WO2010121140; US 8,063,182; US 9,655,964; WO1991003553; WO2001094585; US 9,828,424; US 9,481,736; WO2007024705; and WO2012131053, the contents of each of which are specifically incorporated herein by reference.Anti-IFN-γAntibody Portion

本文所述之抗IFN-γ抗體部分之任一者可用作本文所述之多特異性構築體之任一者的第二、第三或第四抗體部分。Any of the anti-IFN-γ antibody portions described herein can be used as the second, third, or fourth antibody portion of any of the multispecific constructs described herein.

IFN-γ為主要由活化的T細胞及自然殺手(NK)細胞產生的細胞激素，並可促進巨噬細胞活化、媒介抗病毒與抗菌免疫性、增強抗原呈現、協調先天免疫系統的活化、協調淋巴球-內皮細胞相互作用、調節Th1/Th2平衡及控制細胞增生與凋亡。IFN-γ經由活化JAK-STAT路徑促進發炎反應，且為MHC-II產生的主要誘導劑。IFN-γ已被確定為治療自體免疫疾病(諸如乾癬、慢性胃腸道發炎及關節炎)及癌症的可能治療標靶。此示例性抗原可用作特定發炎性疾病(例如，過敏或自體免疫疾病)發展的生物標記、預後指標及抗原表現發炎性疾病的免疫治療標靶。IFN-γ is a cytokine produced primarily by activated T cells and natural killer (NK) cells. It promotes macrophage activation, mediates antiviral and antibacterial immunity, enhances antigen presentation, coordinates activation of the innate immune system, regulates lymphocyte-endothelial cell interactions, modulates the Th1/Th2 balance, and controls cell proliferation and apoptosis. IFN-γ promotes inflammatory responses by activating the JAK-STAT pathway and is a major inducer of MHC-II production. IFN-γ has been identified as a potential therapeutic target for the treatment of autoimmune diseases (such as pityriasis, chronic gastrointestinal inflammation, and arthritis) and cancer. This exemplary antigen can be used as a biomarker for the development of specific inflammatory diseases (e.g., allergy or autoimmune diseases), a prognostic indicator, and a target for immunotherapy of antigen-expressing inflammatory diseases.

在一些實施例中，該抗IFN-γ抗體部分特異性地結合至人類及非人類靈長類動物(諸如食蟹獼猴) IFN-γ。在一些實施例中，該抗IFN-γ抗體部分僅結合至人類IFN-γ。示例性抗IFN-γ抗體及其抗原結合片段包括但不限以下揭露的內容：US 7,700,098；US 7,335,743；US 6,350,860；WO2013078378；WO1994014467；EP0966300；US 4,897,264；EP 240,975；EP 416,652；EP 393,502；EP 369,413；Billiau, A.，Immunol. Today 9, 37-40 (1988)；Hereman, H.等人，J. Exp. Med. 171, 1853-1859 (1990)；Landolfo, S.等人，Science 229, 176-179 (1985)；Didlake, R.H.等人，Transplantation 45, 222-223 (1988)；Jacob, C.O.等人，J. Exp. Med. 166, 789-803 (1987)；及Yong, V.W.等人，Natl. Acad. Sci. USA 88, 7016-7020 (1991)，其各自之內容皆通過引用具體併入本文中。恆定域及Fc域In some embodiments, the anti-IFN-γ antibody portion specifically binds to human and non-human primate (e.g., cynomolgus macaque) IFN-γ. In some embodiments, the anti-IFN-γ antibody portion binds only to human IFN-γ. Exemplary anti-IFN-γ antibodies and antigen-binding fragments thereof include, but are not limited to, those disclosed in US 7,700,098; US 7,335,743; US 6,350,860; WO2013078378; WO1994014467; EP0966300; US 4,897,264; EP 240,975; EP 416,652; EP 393,502; EP 369,413; Billiau, A., Immunol. Today 9, 37-40 (1988); Hereman, H. et al., J. Exp. Med. 171, 1853-1859 (1990); Landolfo, S. et al., Science 229, 176-179 (1985); Didlake, RH et al., Transplantation 45, 222-223 (1988); Jacob, CO et al., J. Exp. Med.166 , 789-803 (1987); and Yong, VW et al., Natl. Acad. Sci. USA 88,7016-7020 (1991 ), the contents of each of which are specifically incorporated herein by reference.

本文進一步描述恆定(例如，CH1、CL)域及Fc域(例如，兩個Fc次單元之每一者)及其變體，其可包括在所述之多特異性構築體或分離的抗IL-13抗體構築體之任一者中。i.恆定域Further described herein are cognate (e.g., CH1, CL) domains and Fc domains (e.g., each of two Fc subunits) and variants thereof, which may be included in any of the multispecific constructs or isolated anti-IL-13 antibody constructs described herein.i.Cognate Domains

恆定域通常被視為負責在可變域與標靶抗原結合後媒介效應功能，諸如調理作用(opsonization)及補體活化。重鏈之恆定域稱為「CH1」，且輕鏈之恆定域稱為「CL」。功能上而言，CH1域賦予效應性質，諸如補體結合、半衰期長度、與FcR的相互作用及Ig之類別或同型。在人類及小鼠中，存在四種IgG，或γ鏈同型，其等對於識別及清除許多肽及多醣抗原至關重要。本文描述了CH1域及/或CL域的突變，該些突變可改善本文所述之多特異性構築體的結構完整性及/或功能。在一些實施例中，該抗IL-13抗體部分包含如本文所述之CH1域及/或CL域的突變。在其他實施例中，該第二抗體部分(例如，抗TSLP抗體部分)包含如本文所述之CH1域及/或CL域的突變。在其他實施例中，該第二抗體部分(例如，抗TSLP抗體部分)及該抗IL-13抗體部分各自包含如本文所述之CH1域及/或CL域的突變。The homeostatic domain is generally considered to be responsible for mediating effector functions, such as opsonization and complement activation, following binding of the variable domain to the target antigen. The homeostatic domain of the heavy chain is called "CH1," and the homeostatic domain of the light chain is called "CL." Functionally, the CH1 domain confers effector properties, such as complement binding, half-life, interaction with FcRs, and class or isotype of Ig. In humans and mice, there are four IgG, or gamma chain isotypes, which are important for recognition and clearance of many polypeptide and polysaccharide antigens. Described herein are mutations in the CH1 domain and/or CL domain that improve the structural integrity and/or function of the multispecific constructs described herein. In some embodiments, the anti-IL-13 antibody portion comprises a mutation in the CH1 domain and/or CL domain as described herein. In other embodiments, the second antibody portion (e.g., an anti-TSLP antibody portion) comprises a mutation in the CH1 domain and/or CL domain as described herein. In other embodiments, the second antibody portion (e.g., an anti-TSLP antibody portion) and the anti-IL-13 antibody portion each comprise a mutation in the CH1 domain and/or CL domain as described herein.

在一些實施例中，本文所述之多特異性構築體之任一者包含第一重鏈(HC1或H1)、第二重鏈(HC2或H2)、第一輕鏈(LC1或L1)及第二(LC2或L2)輕鏈，其中每一HC1及HC2包含人類IgG之重鏈可變域(VH)及第一重鏈恆定域(CH1)，且每一LC1及LC2包含人類IgG之輕鏈可變域(VL)及輕鏈恆定域(CL)。在一些實施例中，該H1包含位於一或多個選自由126、170、183、185及220所組成群組之位置處的取代(EU編號)，且該L1包含位於一或多個選自由124、135及214所組成群組之位置處的取代(EU編號)。在一些實施例中，該H1不包括任何選自由以下所組成群組的取代(EU編號)：L128F、A141M、A141T、A141I、F170M、F170Y、F170S、F170A、S181I、S181T、S181M、S183A、S183V及V185A，且該L1不包括任何選自由以下所組成群組的取代(EU編號)：F116A、F118V、S131T、S131D、V133A、V133I、L135Y、L135V、S162A、S162M、T164S、S174A、S176M、S176A、S176T、S176F、T178L、T178V及T178I。在一些實施例中，該多特異性構築體為人類IgG。在一些實施例中，該多特異性構築體為人類IgG1、IgG2、IgG3或IgG4抗體。在一些實施例中，該多特異性構築體為嵌合抗體。在一些實施例中，該多特異性構築體為人源化抗體。在一些實施例中，該多特異性構築體為嵌合或人源化IgG1、IgG2、IgG3或IgG4抗體。在一些實施例中，該H1包含位於位置F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1之位置F170處的取代(EU編號)為F170V或F170I。在一些實施例中，該H1之位置S183處的取代(EU編號)為S183L或S183I。在一些實施例中，該H1之位置V185處的取代(EU編號)為V185L。在一些實施例中，該L1之位置L135處的取代(EU編號)為L135F。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1之位置F126處的取代(EU編號)為F126C。在一些實施例中，該H1之位置C220處的取代(EU編號)為C220S。在一些實施例中，該L1之位置E124處的取代(EU編號)為E124C。在一些實施例中，該L1之位置Q124處的取代(EU編號)為Q124C。在一些實施例中，該L1之位置C214處的取代(EU編號)為C214S。在一些實施例中，該H1包含一或多個選自由F126C、F170I、S183L、V185L及C220S所組成群組的取代(EU編號)，且該L1包含一或多個選自由Q124C或E124C、L135F及C214S所組成群組的取代(EU編號)。上述針對該H1及該L1的任何取代可改為存在該H2及該L2上。在一些實施例中，該H1及該L1包含上述取代之任一者，且該H2及該L2進一步包含上述針對該H1及該L1的任何取代(可相同或不同)。In some embodiments, any of the multispecific constructs described herein comprises a first heavy chain (HC1 or H1), a second heavy chain (HC2 or H2), a first light chain (LC1 or L1), and a second light chain (LC2 or L2), wherein each HC1 and HC2 comprises a heavy chain variable domain (VH) and a first heavy chain constant domain (CH1) of human IgG, and each LC1 and LC2 comprises a light chain variable domain (VL) and a light chain constant domain (CL) of human IgG. In some embodiments, the H1 comprises a substitution at one or more positions selected from the group consisting of 126, 170, 183, 185, and 220 (EU numbering), and the L1 comprises a substitution at one or more positions selected from the group consisting of 124, 135, and 214 (EU numbering). In some embodiments, the H1 does not include any substitution selected from the group consisting of L128F, A141M, A141T, A141I, F170M, F170Y, F170S, F170A, S181I, S181T, S181M, S183A, S183V, and V185A (EU numbering), and the L1 does not include any substitution selected from Substitutions consisting of the group consisting of (EU numbering): F116A, F118V, S131T, S131D, V133A, V133I, L135Y, L135V, S162A, S162M, T164S, S174A, S176M, S176A, S176T, S176F, T178L, T178V, and T178I. In some embodiments, the multispecific construct is a human IgG. In some embodiments, the multispecific construct is a human IgG1, IgG2, IgG3, or IgG4 antibody. In some embodiments, the multispecific construct is a chimeric antibody. In some embodiments, the multispecific construct is a humanized antibody. In some embodiments, the multispecific construct is a chimeric or humanized IgG1, IgG2, IgG3, or IgG4 antibody. In some embodiments, the H1 comprises substitutions (EU numbering) at positions F170, S183, and V185, and the L1 comprises a substitution (EU numbering) at position L135. In some embodiments, the substitution (EU numbering) at position F170 of the H1 is F170V or F170I. In some embodiments, the substitution (EU numbering) at position S183 of the H1 is S183L or S183I. In some embodiments, the substitution (EU numbering) at position V185 of the H1 is V185L. In some embodiments, the substitution (EU numbering) at position L135 of the L1 is L135F. In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the substitution at position F126 of the H1 (EU numbering) is F126C. In some embodiments, the substitution at position C220 of the H1 (EU numbering) is C220S. In some embodiments, the substitution at position E124 of the L1 (EU numbering) is E124C. In some embodiments, the substitution at position Q124 of L1 (EU numbering) is Q124C. In some embodiments, the substitution at position C214 of L1 (EU numbering) is C214S. In some embodiments, H1 comprises one or more substitutions selected from the group consisting of F126C, F170I, S183L, V185L, and C220S (EU numbering), and L1 comprises one or more substitutions selected from the group consisting of Q124C or E124C, L135F, and C214S (EU numbering). Any of the above substitutions for H1 and L1 may instead be present on H2 and L2. In some embodiments, H1 and L1 comprise any of the above substitutions, and H2 and L2 further comprise any of the above substitutions for H1 and L1 (which may be the same or different).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於F170、S183及V185處的取代(EU編號)，且該L1包含位於L135處的取代(EU編號)。在一些實施例中，F170取代為F170I、S183取代為S183L且V185取代為V185L。在一些實施例中，該第一抗體部分包含含有F170I、S183L及V185L取代(EU編號)的H1及含有L135F取代(EU編號)的L1。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at L135 (EU numbering). In some embodiments, F170 is substituted with F170I, S183 is substituted with S183L, and V185 is substituted with V185L. In some embodiments, the first antibody portion comprises H1 comprising F170I, S183L, and V185L substitutions (EU numbering) and L1 comprising L135F substitution (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain.

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於F170、S183及V185處的取代(EU編號)，且該L1包含位於L135處的取代(EU編號)。在一些實施例中，F170取代為F170V、S183取代為S183I且V185取代為V185L。在一些實施例中，該第一抗體部分包含含有F170V、S183I及V185L取代(EU編號)的H1及含有L135F取代(EU編號)的L1。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at L135 (EU numbering). In some embodiments, F170 is substituted with F170V, S183 is substituted with S183I, and V185 is substituted with V185L. In some embodiments, the first antibody portion comprises H1 comprising F170V, S183I, and V185L substitutions (EU numbering) and L1 comprising L135F substitution (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain.

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中該H1包含位於位置126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein the H1 comprises substitutions at positions 126 and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises F126C and C220S substitutions (EU numbering), and the L1 comprises E124C and C214S substitutions (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises F126C and C220S substitutions (EU numbering), and the L1 comprises Q124C and C214S substitutions (EU numbering).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置126、170、183、185及220處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 126, 170, 183, 185, and 220 (EU numbering), and the L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions (EU numbering), and the L1 comprises E124C, L135F, and C214S substitutions (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions (EU numbering), and the L1 comprises Q124C, L135F, and C214S substitutions (EU numbering).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置126、170、183、185及220處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。ii. Fc域In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 126, 170, 183, 185, and 220 (EU numbering), and the L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions (EU numbering), and the L1 comprises E124C, L135F, and C214S substitutions (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions (EU numbering), and the L1 comprises Q124C, L135F, and C214S substitutions (EU numbering).ii. Fcdomain

Fc (片段、可結晶)域為抗體的尾部區域，其與稱為Fc受體的細胞表面受體及補體系統的一些蛋白質相互作用。抗體之Fc媒介的效應功能包括抗體依賴性細胞毒性(ADCC)、抗體依賴性細胞吞噬作用(ADCP)及補體依賴性細胞毒性(CDC)。此性質為抗體活化免疫系統的關鍵。該域由二或三個恆定域組成，該些恆定域附接至兩條重鏈之每一者上的CH結構域，取決於抗體的類別。當組裝兩條抗體多肽時，兩條重鏈上的Fc域之CH3次單元的突變(稱為「杵臼」突變)可進一步改善異二聚合作用。舉例而言，當以下的「臼」取代被導入第二重鏈上的Fc域之CH3域的位置366、368及407處：T366S、L368A及Y407V時，第一重鏈上的Fc域之CH3域可包含「杵」突變(例如，T366W)。杵與臼突變的位置可轉換，例如該杵突變可位於第二重鏈上，且該臼突變可位於第一重鏈上。可導入額外的Fc域突變，諸如LALA突變(例如，Fc域之CH2域的L234A及L235A取代)，以降低FcγR活化及Fc媒介的毒性，其可在投予該抗體之個體中引發副作用，諸如細胞激素釋放症候群。此外，可導入Fc域突變，諸如LS突變(例如，Fc域之CH3域的M428L及N434S取代)，以延長抗體半衰期而不影響效力。本文亦可使用FcRn親和力增強Fc突變體，諸如M252Y/S254T/T256E (YTE)突變，其在導入Fc域中時，能夠延長血清半衰期。因此，所述之突變可單獨或組合使用，以改善抗體功能及降低對患者的毒性。在一些實施例中，該Fc域包含LALA突變。在一些實施例中，該Fc域包含LS或YTE突變。在一些實施例中，該Fc域包含LALA突變及LS 突變。在一些實施例中，該Fc域包含LALA突變及YTE突變。在一些實施例中，i)該Fc域之第一次單元及第二次單元各自含有SEQ ID NO:76-79之任一者的胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；ii)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；iii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；iv)該Fc域之第一次單元含有SEQ ID NO:95之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:96之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段；或v)該Fc域之第一次單元含有SEQ ID NO:96之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段，且該Fc域之第二次單元含有SEQ ID NO:95之胺基酸序列或其缺乏SEQ ID NO:215之肽序列的片段。The Fc (fragment, crystallizable) domain is the tail region of an antibody that interacts with cell surface receptors called Fc receptors and certain proteins of the complement system. The Fc-mediated effector functions of antibodies include antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and complement-dependent cytotoxicity (CDC). This property is key to the ability of antibodies to activate the immune system. This domain is composed of two or three constant domains attached to the CH domain on each of the two heavy chains, depending on the class of antibody. When two antibody polypeptides are assembled, mutations in the CH3 subunits of the Fc domains on both heavy chains (called "knob-in-hole" mutations) can further improve heterodimerization. For example, the CH3 domain of the Fc domain on the first heavy chain may comprise a "knob" mutation (e.g., T366W) when the following "hole" substitutions are introduced into the CH3 domain of the Fc domain on the second heavy chain at positions 366, 368, and 407: T366S, L368A, and Y407V. The locations of the knob and hole mutations can be reversed, e.g., the knob mutation can be located on the second heavy chain and the hole mutation can be located on the first heavy chain. Additional Fc domain mutations, such as LALA mutations (e.g., L234A and L235A substitutions in the CH2 domain of the Fc domain), can be introduced to reduce FcγR activation and Fc-mediated toxicity, which can cause side effects, such as cytokine release syndrome, in subjects administered the antibody. In addition, Fc domain mutations, such as LS mutations (e.g., M428L and N434S substitutions in the CH3 domain of the Fc domain), can be introduced to extend antibody half-life without affecting efficacy. FcRn affinity enhancing Fc mutants, such as M252Y/S254T/T256E (YTE) mutations, can also be used herein, which, when introduced into the Fc domain, can extend serum half-life. Thus, the mutations described can be used alone or in combination to improve antibody function and reduce toxicity to patients. In some embodiments, the Fc domain comprises a LALA mutation. In some embodiments, the Fc domain comprises a LS or YTE mutation. In some embodiments, the Fc domain comprises a LALA mutation and a LS mutation. In some embodiments, the Fc domain comprises a LALA mutation and a YTE mutation. In some embodiments, i) the first and second Fc domain units each contain the amino acid sequence of any one of SEQ ID NOs: 76-79 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; ii) the first Fc domain unit contains the amino acid sequence of SEQ ID NO: 80 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215, and the second Fc domain unit contains the amino acid sequence of SEQ ID NO: 81 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; iii) the first Fc domain unit contains the amino acid sequence of SEQ ID NO: 81 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215, and the second Fc domain unit contains the amino acid sequence of SEQ ID NO: 80 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; iv) the first Fc domain unit contains the amino acid sequence of SEQ ID NO: 95 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 96. NO: 215, and the second Fc domain subunit contains the amino acid sequence of SEQ ID NO: 96 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215; or v) the first Fc domain subunit contains the amino acid sequence of SEQ ID NO: 96 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215, and the second Fc domain subunit contains the amino acid sequence of SEQ ID NO: 95 or a fragment thereof lacking the peptide sequence of SEQ ID NO: 215.

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置L234、L235、F170、S183及V185處的取代(EU編號)，且該L1包含位於L135處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A、Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234, L235, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at L135 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises L135F substitutions (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置L234、L235、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234, L235, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, F170V, S183I, and V185L substitutions (EU numbering), and the L1 comprises L135F substitutions (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 126, and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, F126C, and C220S substitutions (EU numbering), and the L1 comprises E124C and C214S substitutions (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, F126C, and C220S substitutions (EU numbering), and the L1 comprises Q124C and C214S substitutions (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、126、170、183、185及220處的取代(EU編號)；且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C或Q124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於L234、L235、F126、F170、S183、V185及C220位置處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: H1 comprises substitutions at positions 234, 235, 126, 170, 183, 185, and 220 (EU numbering); and L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C or Q124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、126、170、183、220及185處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、F126、F170、S183、C220及V185處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、F126、F170、S183、C220及V185處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 126, 170, 183, 220, and 185 (EU numbering), and the L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, F126, F170, S183, C220, and V185 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, F126, F170, S183, C220, and V185 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置428、434、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置M428、N434、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含M428L、N434S、F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 428, 434, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions M428, N434, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises M428L, N434S, F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises L135F substitutions (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置428、434、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置M428、N434、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含M428L、N434S、F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 428, 434, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions M428, N434, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises M428L, N434S, F170V, S183I, and V185L substitutions (EU numbering), and the L1 comprises L135F substitutions (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置428、434、126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置M428、N434、F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含M428L、N434S、F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置M428、N434、F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含M428L、N434S、F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 428, 434, 126, and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions M428, N434, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises M428L, N434S, F126C, and C220S substitutions (EU numbering), and the L1 comprises E124C and C214S substitutions (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions M428, N434, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises M428L, N434S, F126C, and C220S substitutions (EU numbering), and the L1 comprises Q124C and C214S substitutions (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置428、434、126、170、183、185及220處的取代(EU編號)；且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含M428L、N434S、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含M428L、N434S、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: H1 comprises substitutions at positions 428, 434, 126, 170, 183, 185, and 220 (EU numbering); and L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions M428L, N434S, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions M428L, N434S, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置428、434、126、170、183、220及185處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置M428、N434、F126、F170、S183、C220及V185處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含M428L、N434S、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置M428、N434、F126、F170、S183、C220及V185處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含M428L、N434S、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1 and a first subunit of an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 428, 434, 126, 170, 183, 220, and 185 (EU numbering), and the L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions M428, N434, F126, F170, S183, C220, and V185 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions M428L, N434S, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions M428, N434, F126, F170, S183, C220, and V185 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions M428L, N434S, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置252、254、256、170、183及185處的取代(EU編號)；且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置M252、S254、T256、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含M252Y、S254T、T256E、F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 252, 254, 256, 170, 183, and 185 (EU numbering); and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions M252, S254, T256, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, H1 comprises M252Y, S254T, T256E, F170I, S183L, and V185L substitutions (EU numbering), and L1 comprises L135F substitutions (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置252、254、256、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置M252、S254、T256、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含M252Y、S254T、T256E、F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 252, 254, 256, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions M252, S254, T256, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, H1 comprises M252Y, S254T, T256E, F170V, S183I, and V185L substitutions (EU numbering), and L1 comprises L135F substitutions (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置252、254、256、126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置M252、S254、T256、F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含M252Y、S254T、T256E、F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置處的取代M252、S254、T256、F126及C220 (EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含M252Y、S254T、T256E、F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 252, 254, 256, 126, and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions M252, S254, T256, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises M252Y, S254T, T256E, F126C, and C220S substitutions (EU numbering), and the L1 comprises E124C and C214S substitutions (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions M252, S254, T256, F126, and C220 at positions (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises M252Y, S254T, T256E, F126C, and C220S substitutions (EU numbering), and the L1 comprises Q124C and C214S substitutions (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置代252、254、256、126、170、183、185及220處的取(EU編號)；且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置M252、S254、T256、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含M252Y、S254T、T256E、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置M252、S254、T256、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含M252Y、S254T、T256E、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: H1 comprises substitutions at positions 252, 254, 256, 126, 170, 183, 185, and 220 (EU numbering); and L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions M252, S254, T256, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions M252Y, S254T, T256E, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions M252, S254, T256, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions M252Y, S254T, T256E, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置252、254、256、126、170、183、220及185處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置M252、S254、T256、F126、F170、S183、C220及V185處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含M252Y、S254T、T256E、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置M252、S254、T256、F126、F170、S183、C220及V185處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含M252Y、S254T、T256E、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: H1 comprises substitutions at positions 252, 254, 256, 126, 170, 183, 220, and 185 (EU numbering), and L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions M252, S254, T256, F126, F170, S183, C220, and V185 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions M252Y, S254T, T256E, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions M252, S254, T256, F126, F170, S183, C220, and V185 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions M252Y, S254T, T256E, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、428、434、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置L234、L235、M428、N434、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M428L、N434S、F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 428, 434, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234, L235, M428, N434, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, M428L, N434S, F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises L135F substitutions (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、428、434、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置L234、L235、M428、N434、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M428L、N434S、F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 428, 434, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234, L235, M428, N434, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, H1 comprises L234A, L235A, M428L, N434S, F170V, S183I, and V185L substitutions (EU numbering), and L1 comprises L135F substitutions (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、428、434、126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M428、N434、F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M428L、N434S、F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M428、N434、F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M428L、N434S、F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 428, 434, 126, and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M428, N434, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, M428L, N434S, F126C, and C220S substitutions (EU numbering), and the L1 comprises E124C and C214S substitutions (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M428, N434, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, M428L, N434S, F126C, and C220S substitutions (EU numbering), and the L1 comprises Q124C and C214S substitutions (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、428、434、126、170、183、185及220處的取代(EU編號)；且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M428L、N434S、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M428L、N434S、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: H1 comprises substitutions at positions 234, 235, 428, 434, 126, 170, 183, 185, and 220 (EU numbering); and L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M428L, N434S, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M428L, N434S, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、428、434、126、170、183、220及185處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M428L、N434S、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M428L、N434S、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1 and a first subunit of an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 428, 434, 126, 170, 183, 220, and 185 (EU numbering), and the L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M428L, N434S, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M428L, N434S, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、170、183及185處的取代(EU編號)；且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, multispecific constructs described herein are provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 252, 254, 256, 170, 183, and 185 (EU numbering); and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, M252Y, S254T, T256E, F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises L135F substitutions (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 252, 254, 256, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, H1 comprises L234A, L235A, M252Y, S254T, T256E, F170V, S183I, and V185L substitutions (EU numbering), and L1 comprises L135F substitutions (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 252, 254, 256, 126, and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, F126C, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, F126C, and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、126、170、183、185及220處的取代(EU編號)；且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: H1 comprises substitutions at positions 234, 235, 252, 254, 256, 126, 170, 183, 185, and 220 (EU numbering); and L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、126、170、183、220及185處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 252, 254, 256, 126, 170, 183, 220, and 185 (EU numbering), and the L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C, L135F, and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、428、434、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、M428、N434、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、M428L、N434S、F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1 and a first subunit of an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 252, 254, 256, 428, 434, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, M428, N434, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, M428L, N434S, F170I, S183L, and V185L (EU numbering), and the L1 comprises a substitution at position L135F (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、428、434、170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、M428、N434、F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、M428L、N434S、F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1 and a first subunit of an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 252, 254, 256, 428, 434, 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, M428, N434, F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, M428L, N434S, F170V, S183I, and V185L (EU numbering), and the L1 comprises a substitution at position L135F (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、428、434、126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、M428、N434、F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、M428L、N434S、F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、M428、N434、F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、M428L、N434S、F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1 and a first subunit of an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 252, 254, 256, 428, 434, 126, and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, M428, N434, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, M428L, N434S, F126C, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, M428, N434, F126, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, M428L, N434S, F126C, and C220S (EU numbering), and the L1 comprises substitutions at positions Q124C and C214S (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、428、434、126、170、183、185及220處的取代(EU編號)；且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、M428L、N434S、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、M428L、N434S、F126C、F170I、S183L、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1, a first subunit comprising an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: H1 comprises substitutions at positions 234, 235, 252, 254, 256, 428, 434, 126, 170, 183, 185, and 220 (EU numbering); and L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, M428L, N434S, F126C, F170I, S183L, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, M252Y, S254T, T256E, M428L, N434S, F126C, F170I, S183L, V185L, and C220S substitutions (EU numbering), and the L1 comprises Q124C, L135F, and C214S substitutions (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).

在一些實施例中，提供本文所述之多特異性構築體，其包含第一抗體部分，該第一抗體部分包含含有VH1及H1-CH1的第一重鏈(H1)及Fc域之第一次單元以及含有VL1及L1-CL的第一輕鏈(L1)，其中：該H1包含位於位置234、235、252、254、256、428、434、126、170、183、220及185處的取代(EU編號)，且該L1包含位於位置124、135及214處的取代(EU編號)。在一些實施例中，該L1係衍生自λ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置E124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、M428L、N434S、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含E124C、L135F及C214S取代(EU編號)。在一些實施例中，該L1係衍生自κ輕鏈。在一些實施例中，該H1包含位於位置L234、L235、M252、S254、T256、M428、N434、F126、F170、S183、V185及C220處的取代(EU編號)，且該L1包含位於位置Q124、L135及C214處的取代(EU編號)。在一些實施例中，該H1包含L234A、L235A、M252Y、S254T、T256E、M428L、N434S、F126C、F170V、S183I、V185L及C220S取代(EU編號)，且該L1包含Q124C、L135F及C214S取代(EU編號)。在一些實施例中，本文所述之多特異性構築體進一步包含第二抗體部分，該第二抗體部分包含含有VH2及H2-CH1的第二重鏈(H2)及Fc域之第二次單元以及含有VL2及L2-CL的第二輕鏈(L2)；其中(i)該H1進一步包含T366W取代(EU編號)，且該H2包含T366S、L368A及Y407V取代(EU編號)；或(ii)該H1進一步包含T366S、L368A及Y407V取代(EU編號)，且該H2包含T366W取代(EU編號)。在一些實施例中，該L2係衍生自κ輕鏈。在一些實施例中，該L2係衍生自λ輕鏈。在一些實施例中，該Fc域係衍生自IgG1 (例如，人類IgG1)。連接子In some embodiments, a multispecific construct described herein is provided, comprising a first antibody portion comprising a first heavy chain (H1) comprising VH1 and H1-CH1 and a first subunit of an Fc domain, and a first light chain (L1) comprising VL1 and L1-CL, wherein: the H1 comprises substitutions at positions 234, 235, 252, 254, 256, 428, 434, 126, 170, 183, 220, and 185 (EU numbering), and the L1 comprises substitutions at positions 124, 135, and 214 (EU numbering). In some embodiments, the L1 is derived from a lambda light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions E124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises substitutions at positions L234A, L235A, M252Y, S254T, T256E, M428L, N434S, F126C, F170V, S183I, V185L, and C220S (EU numbering), and the L1 comprises substitutions at positions E124C, L135F, and C214S (EU numbering). In some embodiments, the L1 is derived from a kappa light chain. In some embodiments, the H1 comprises substitutions at positions L234, L235, M252, S254, T256, M428, N434, F126, F170, S183, V185, and C220 (EU numbering), and the L1 comprises substitutions at positions Q124, L135, and C214 (EU numbering). In some embodiments, the H1 comprises L234A, L235A, M252Y, S254T, T256E, M428L, N434S, F126C, F170V, S183I, V185L, and C220S substitutions (EU numbering), and the L1 comprises Q124C, L135F, and C214S substitutions (EU numbering). In some embodiments, the multispecific constructs described herein further comprise a second antibody portion comprising a second heavy chain (H2) comprising VH2 and H2-CH1, a second subunit of an Fc domain, and a second light chain (L2) comprising VL2 and L2-CL; wherein (i) the H1 further comprises a T366W substitution (EU numbering), and the H2 comprises T366S, L368A, and Y407V substitutions (EU numbering); or (ii) the H1 further comprises T366S, L368A, and Y407V substitutions (EU numbering), and the H2 comprises a T366W substitution (EU numbering). In some embodiments, the L2 is derived from a kappa light chain. In some embodiments, the L2 is derived from a lambda light chain. In some embodiments, the Fc domain is derived from IgG1 (e.g., human IgG1).connector

本文所述之多特異性抗體構築體可包含連接本文所述之任何多特異性抗體構築體中之任何Fab或scFv片段之VH或VL的連接子(諸如肽連接子)。在一些實施例中，該連接子連接Fab與全長抗體之第一鏈或第二鏈。在一些實施例中，該連接子連接scFv與全長抗體之第一鏈或第二鏈。在一些實施例中，該連接子連接Fab片段與scFv片段。在一些實施例中，該連接子連接scFv片段之VH與VL。在一些實施例中，該多特異性構築體不包含連接子。在一些實施例中，該多特異性構築體包含1個連接子。在一些實施例中，該多特異性構築體包含2或多個連接子(例如，2、3、4或多個連接子之任一者)。在一些實施例中，該第一連接子與該第二連接子相同。在一些實施例中，該第一連接子與該第二連接子不同。The multispecific antibody constructs described herein may comprise a linker (e.g., a peptide linker) that connects the VH or VL of any Fab or scFv fragment in any multispecific antibody construct described herein. In some embodiments, the linker connects the Fab to the first or second chain of the full-length antibody. In some embodiments, the linker connects the scFv to the first or second chain of the full-length antibody. In some embodiments, the linker connects the Fab fragment to the scFv fragment. In some embodiments, the linker connects the VH and VL of the scFv fragment. In some embodiments, the multispecific construct does not comprise a linker. In some embodiments, the multispecific construct comprises one linker. In some embodiments, the multispecificity construct comprises two or more linkers (e.g., any of two, three, four, or more linkers). In some embodiments, the first linker is the same as the second linker. In some embodiments, the first linker is different from the second linker.

該連接子可為任何長度的肽連接子。在一些實施例中，該肽連接子長度約1至約10個胺基酸、長度約2至約15個胺基酸、長度約3至約12個胺基酸、長度約4至約10個胺基酸、長度約5至約9個胺基酸、長度約6至約8個胺基酸、長度約1至約20個胺基酸、長度約21至約30個胺基酸、長度約1至約30個胺基酸、長度約2至約20個胺基酸、長度約10至約30個胺基酸、長度約2至約19個胺基酸、長度約2至約18個胺基酸、長度約2至約17個胺基酸、長度約2至約16個胺基酸、長度約2至約10個胺基酸、長度約2至約14個胺基酸、長度約2至約13個胺基酸、長度約2至約12個胺基酸、長度約2至約11個胺基酸、長度約2至約9個胺基酸、長度約2至約8個胺基酸、長度約2至約7個胺基酸、長度約2至約6個胺基酸或長度約2至約5個胺基酸。在一些實施例中，該肽連接子為長度1、2、3、4、5、6、7、8、9、10、11、12、13、14、15、16、17、18、19或20個胺基酸之任一者。在一些實施例中，該肽連接子長度21、22、23、24、25、26、27、28、29或30個胺基酸之任一者。在一些實施例中，該肽連接子長度約2至約30個胺基酸，諸如長度約2至約15個胺基酸、長度約15個胺基酸或長度約6個胺基酸。舉例而言，在一些實施例中，該肽連接子的N端係共價連接至全長抗TSLP抗體的C端，且該肽連接子的C端係共價連接至抗IL-13 Fab片段之VH或VL的N端。The linker can be a peptide linker of any length. In some embodiments, the peptide linker is about 1 to about 10 amino acids in length, about 2 to about 15 amino acids in length, about 3 to about 12 amino acids in length, about 4 to about 10 amino acids in length, about 5 to about 9 amino acids in length, about 6 to about 8 amino acids in length, about 1 to about 20 amino acids in length, about 21 to about 30 amino acids in length, about 1 to about 30 amino acids in length, about 2 to about 20 amino acids in length, about 10 to about 30 amino acids in length, about 2 to about 19 amino acids in length, or about 1 to about 20 amino acids in length. In some embodiments, the peptide linker is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 amino acids in length. In some embodiments, the peptide linker is any of 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 amino acids in length. In some embodiments, the peptide linker is about 2 to about 30 amino acids in length, such as about 2 to about 15 amino acids in length, about 15 amino acids in length, or about 6 amino acids in length. For example, in some embodiments, the N-terminus of the peptide linker is covalently linked to the C-terminus of the full-length anti-TSLP antibody, and the C-terminus of the peptide linker is covalently linked to the N-terminus of the VH or VL of the anti-IL-13 Fab fragment.

肽連接子可具有天然存在的序列或非天然存在的序列。舉例而言，衍生自僅抗體重鏈鉸鏈區之序列可用作連接子。參見例如WO1996/34103。在一些實施例中，該肽連接子為人類IgG1或IgG4鉸鏈。在一些實施例中，該肽連接子為突變的人類IgG1或IgG4鉸鏈。在一些實施例中，該連接子為撓性連接子。示例性撓性連接子包括甘胺酸聚合物(G)n (例如，GG)、甘胺酸-絲胺酸聚合物(包括例如：(GS)n、(GSGGS)n (SEQ ID NO:14)、(GGGS)n (SEQ ID NO:16)或(GGGGS)n (SEQ ID NO:17)，其中n為至少1的整數)、甘胺酸-丙胺酸聚合物、丙胺酸-絲胺酸聚合物及本領域已知之其他撓性連接子。甘胺酸及甘胺酸-絲胺酸聚合物相對非結構化，從而可充當各組分之間的中性栓鏈(tether)。甘胺酸比丙胺酸明顯擷取更多的phi-psi空間，且比側鏈較長的殘基更不受限制(參見Scheraga，Rev. Computational Chem. 11 173-142 (1992))。在一些實施例中，該連接子包含選自由以下所組成群組之胺基酸：甘胺酸、絲胺酸、精胺酸及丙胺酸。示例性撓性連接子包括但不限於：Gly-Gly、Gly-Gly-Ser-Gly (SEQ ID NO:18)、Gly-Gly-Ser-Gly-Gly (SEQ ID NO:163)、Gly-Ser-Gly-Ser-Gly (SEQ ID NO:164)、Gly-Ser-Gly-Gly-Gly (SEQ ID NO:165)、Gly-Gly-Gly-Ser-Gly (SEQ ID NO:166)、Gly-Ser-Ser-Ser-Gly (SEQ ID NO:167)、Gly-Gly-Ser-Gly-Gly-Ser (SEQ ID NO:168)、Ser-Gly-Gly-Gly-Gly-Ser (SEQ ID NO:169)、Gly-Arg-Ala-Gly-Gly-Gly-Gly-Ala-Gly-Gly-Gly-Gly (SEQ ID NO:170)、Gly-Arg-Ala-Gly-Gly-Gly (SEQ ID NO:171)、GGGGSGGGGSGGGGS (SEQ ID NO:99)、GGGGS (SEQ ID NO:172)、AGGGGSGGGGSGGGGS (SEQ ID NO:224)及其類似物。在一些實施例中，介於(例如)該抗TSLP Fab片段與該抗IL-13 scFv片段之間的連接子為GGGG (SEQ ID NO: 98)。在一些實施例中，介於該抗IL-13 scFv片段或該抗TSLP scFv片段哦VH與VL之間的連接子為GGGGSGGGGSGGGGS (SEQ ID NO:99)。在一些實施例中，連接該Fc域與該抗IL-13 scFv或抗TSLP scFv的連接子為GG或GGGG (SEQ ID NO:98)。具有通常知識者將理解，多特異性構築體之設計可包括全部或部分撓性的連接子，使得該連接子可包括撓性連接子部分以及一或多個賦予較少撓性結構的部分，以提供所需之多特異性構築體結構。The peptide linker can have a naturally occurring sequence or a non-naturally occurring sequence. For example, a sequence derived from the hinge region of an antibody heavy chain can be used as a linker. See, for example, WO 1996/34103. In some embodiments, the peptide linker is a human IgG1 or IgG4 hinge. In some embodiments, the peptide linker is a mutated human IgG1 or IgG4 hinge. In some embodiments, the linker is a flexible linker. Exemplary flexible linkers include glycine polymers (G)n (e.g., GG), glycine-serine polymers (including, for example, (GS)n, (GSGGS)n (SEQ ID NO: 14), (GGGS)n (SEQ ID NO: 16), or (GGGGS)n (SEQ ID NO: 17), where n is an integer of at least 1), glycine-alanine polymers, alanine-serine polymers, and other flexible linkers known in the art. Glycine and glycine-serine polymers are relatively unstructured and thus can serve as neutral tethers between the components. Glycine takes up significantly more phi-psi space than alanine and is less constrained than longer side chain residues (see Scheraga, Rev. Computational Chem. 11 173-142 (1992)). In some embodiments, the linker comprises an amino acid selected from the group consisting of glycine, serine, arginine, and alanine. Exemplary flexible linkers include, but are not limited to: Gly-Gly, Gly-Gly-Ser-Gly (SEQ ID NO: 18), Gly-Gly-Ser-Gly-Gly (SEQ ID NO: 163), Gly-Ser-Gly-Ser-Gly (SEQ ID NO: 164), Gly-Ser-Gly-Gly-Gly (SEQ ID NO: 164) NO:165), Gly-Gly-Gly-Ser-Gly (SEQ ID NO:166), Gly-Ser-Ser-Ser-Gly (SEQ ID NO:167), Gly-Gly-Ser-Gly-Gly-Ser (SEQ ID NO:168), Ser-Gly-Gly-Gly-Gly-Ser (SEQ ID NO: 169), Gly-Arg-Ala-Gly-Gly-Gly-Gly-Ala-Gly-Gly-Gly-Gly (SEQ ID In some embodiments, the linker between the VH and VL of the anti-IL-13 scFv fragment or the anti-TSLP scFv fragment is GGGGSGGGGSGGGGS (SEQ ID NO: 99). In some embodiments, the linker connecting the Fc domain to the anti-IL-13 scFv or anti-TSLP scFv is GG or GGGG (SEQ ID NO: 98). One of ordinary skill in the art will appreciate that the design of a multispecific construct can include a fully or partially flexible linker, such that the linker can include a flexible linker portion and one or more portions that impart less flexible structure to provide the desired multispecific construct structure.

在一些實施例中，該第二抗體部分(例如，抗TSLP抗體部分)與該抗IL-13抗體部分之間的連接子，或該第二抗體部分(例如，抗TSLP抗體部分)及/或該抗IL-13抗體部分之內的連接子為穩定的連接子(不被蛋白酶(尤其是MMP)裂解)。In some embodiments, the linker between the second antibody portion (e.g., anti-TSLP antibody portion) and the anti-IL-13 antibody portion, or the linker within the second antibody portion (e.g., anti-TSLP antibody portion) and/or the anti-IL-13 antibody portion is a stable linker (not cleaved by proteases, particularly MMPs).

在一些實施例中，該連接子可裂解的連接子。在一些實施例中，該第二抗體部分(例如，抗TSLP抗體部分)之VH或VL與該抗IL-13抗體部分之間的連接子包含蛋白酶受質裂解序列，例如MMP受質裂解序列。可被MMP裂解的受質序列已被廣泛研究。舉例而言，PLGLAG之序列(SEQ ID NO:97)可被大多數的MMP裂解。在一些實施例中，蛋白酶裂解位點可被MMP-2、MMP-9或其組合辨識。In some embodiments, the linker is a cleavable linker. In some embodiments, the linker between the VH or VL of the second antibody portion (e.g., the anti-TSLP antibody portion) and the anti-IL-13 antibody portion comprises a protease substrate cleavage sequence, such as a MMP substrate cleavage sequence. Substrate sequences cleavable by MMPs have been extensively studied. For example, the sequence of PLGLAG (SEQ ID NO: 97) is cleavable by most MMPs. In some embodiments, the protease cleavage site is recognized by MMP-2, MMP-9, or a combination thereof.

在一些實施例中，該多特異性構築體(諸如雙特異性構築體)包含各自含有以下結構的第一及第二多肽：N’ - 抗TSLP全長抗體 - L1 - 抗IL-13 VL- L2 - 抗IL-13 VH - C’。在一些實施例中，該多特異性構築體包含各自含有以下結構的第一及第二多肽：N’ - 抗TSLP VH1 - (H1-CH1) - L1 - 抗IL-13 VL1 - L2 - 抗IL-13 VH1 - L3 - Fc域 - C’，以及含有以下結構的第三及第四多肽：N’ - 抗TSLP VL3 - (L3-CL) - C’。在一些實施例中，本文所述之多特異性構築體包含含有以下結構的第一及第二多肽：N’ - 抗IL-13全長抗體 - L1 - 抗TSLP VH - L2 - 抗TSLP VL - C’。在一些實施例中，該多特異性構築體包含各自含有以下結構的第一及第二多肽：N’ - 抗IL-13 VH1 - (H1-CH1) - L1 - 抗TSLP VH1 - L2 - 抗TSLP VL1 - L3 - Fc域 - C’，以及含有以下結構的第三及第四多肽：N’ - 抗IL-13 VL2 - (L2-CL) - C’。在一些實施例中，該多特異性構築體包含各自含有以下結構的第一及第二多肽：N’ - 抗TSLP全長抗體 - L1 - 抗IL-13 VH1 - (H1-CH1) - C’，其中L1連接該抗TSLP全長抗體的C’端與該抗IL-13 Fab的N端，以及各自含有以下結構的第三及第四多肽：N’ - 抗IL-13 VL1-(L1-CL) - C’。在一些實施例中，該多特異性構築體包含各自含有以下結構的第一及第二多肽：N’ - 抗IL-13全長抗體 - L1 - 抗TSLP VH1 - (H1-CH1) - C’，其中L1連接該抗IL-13全長抗體的C’端與該抗TSLP Fab的N端，以及各自含有以下結構的第三及第四多肽：N’ - 抗TSLP VL1 - (L1-CL) - C’。在一些實施例中，該多特異性構築體包含各自含有以下結構的第一及第二多肽：N’ - 抗IL-13 VH1-(H1-CH1) - L1 - 抗TSLP全長抗體 - C’，以及各自含有以下結構的第三及第四多肽：N’ - 抗IL-13 VL1 - (L1-CL) - C’。在一些實施例中，該多特異性構築體包含含有以下結構的第一多肽：N’ - 抗TSLP VH1 - (H1-CH1) - L1 - 抗TSLP全長抗體 - C’，含有以下結構的第二多肽：N’ - 抗TSLP VL1 - (L1-CL) - C’，含有以下結構的第三多肽：N’ - 抗IL-13 VH1 - (H1-CH1) - L1 - 抗IL-13全長抗體 - C’，以及含有以下結構的第四多肽：N’ - 抗IL-13 VL1 - (L1-CL) - C’。在一些實施例中，該多特異性構築體包含含有以下結構的第一多肽：N’ - 抗TSLP VH1 - (H1-CH1) - Fc域 - C’，含有以下結構的第二多肽：N’ - 抗TSLP VL1 - (L1-CL) - C’，含有以下結構的第三多肽：N’ - 抗IL-13 VH1 - (H1-CH1) - Fc域 - C’，以及含有以下結構的第四多肽：N’ - 抗IL-13 VL1 - (L1-CL) - C’。L1、L2及L3為本文所述之任何可能的連接子(諸如肽連接子)。L1-L3可相同或不同。III.製備方法In some embodiments, the multispecific construct (e.g., a bispecific construct) comprises first and second polypeptides, each comprising the following structure: N'-anti-TSLP full-length antibody-L1-anti-IL-13 VL-L2-anti-IL-13 VH-C'. In some embodiments, the multispecific construct comprises first and second polypeptides, each comprising the following structure: N'-anti-TSLP VH1-(H1-CH1)-L1-anti-IL-13 VL1-L2-anti-IL-13 VH1-L3-Fc domain-C', and third and fourth polypeptides comprising the following structure: N'-anti-TSLP VL3-(L3-CL)-C'. In some embodiments, the multispecific constructs described herein comprise first and second polypeptides having the following structure: N'-anti-IL-13 full-length antibody-L1-anti-TSLP VH-L2-anti-TSLP VL-C'. In some embodiments, the multispecific constructs comprise first and second polypeptides each having the following structure: N'-anti-IL-13 VH1-(H1-CH1)-L1-anti-TSLP VH1-L2-anti-TSLP VL1-L3-Fc domain-C', and third and fourth polypeptides having the following structure: N'-anti-IL-13 VL2-(L2-CL)-C'. In some embodiments, the multispecific construct comprises first and second polypeptides, each comprising the following structure: N'-anti-TSLP full-length antibody-L1-anti-IL-13 VH1-(H1-CH1)-C', wherein L1 connects the C'-terminus of the anti-TSLP full-length antibody and the N-terminus of the anti-IL-13 Fab, and third and fourth polypeptides, each comprising the following structure: N'-anti-IL-13 VL1-(L1-CL)-C'. In some embodiments, the multispecific construct comprises first and second polypeptides, each comprising the following structure: N'-anti-IL-13 full-length antibody-L1-anti-TSLP VH1-(H1-CH1)-C', wherein L1 connects the C' terminus of the anti-IL-13 full-length antibody to the N-terminus of the anti-TSLP Fab, and third and fourth polypeptides, each comprising the following structure: N'-anti-TSLP VL1-(L1-CL)-C'. In some embodiments, the multispecific construct comprises first and second polypeptides, each comprising the following structure: N'-anti-IL-13 VH1-(H1-CH1)-L1-anti-TSLP full-length antibody-C', and third and fourth polypeptides, each comprising the following structure: N'-anti-IL-13 VL1-(L1-CL)-C'. In some embodiments, the multispecific construct comprises a first polypeptide comprising the following structure: N'-anti-TSLP VH1-(H1-CH1)-L1-anti-TSLP full-length antibody-C', a second polypeptide comprising the following structure: N'-anti-TSLP VL1-(L1-CL)-C', a third polypeptide comprising the following structure: N'-anti-IL-13 VH1-(H1-CH1)-L1-anti-IL-13 full-length antibody-C', and a fourth polypeptide comprising the following structure: N'-anti-IL-13 VL1-(L1-CL)-C'. In some embodiments, the multispecific construct comprises a first polypeptide having the following structure: N'-anti-TSLP VH1-(H1-CH1)-Fc domain-C', a second polypeptide having the following structure: N'-anti-TSLP VL1-(L1-CL)-C', a third polypeptide having the following structure: N'-anti-IL-13 VH1-(H1-CH1)-Fc domain-C', and a fourth polypeptide having the following structure: N'-anti-IL-13 VL1-(L1-CL)-C'. L1, L2, and L3 are any possible linkers (e.g., peptide linkers) described herein. L1-L3 may be the same or different.III.Preparation Methods

亦提供製造本文所述之抗IL-13抗體構築體及/或多特異性構築體之任一者的方法。Also provided are methods of making any of the anti-IL-13 antibody constructs and/or multispecific constructs described herein.

本文所述之抗IL-13抗體構築體及/或多特異性構築體可各自由本領域已知之任何蛋白質表現及純化方法來製備。可完全合成編碼抗IL-13抗體構築體及/或多特異性構築體的DNA序列。在獲得此類序列之後，將其選殖至適合的表現載體中，隨後轉染至適合的宿主細胞中。培養轉染的宿主細胞，收穫並純化上清液，以獲得本文所述之抗IL-13抗體構築體或多特異性構築體。在一些實施例中，將宿主細胞溶解，以獲得表現的多特異性構築體或分離的抗TSLP抗體構築體。The anti-IL-13 antibody constructs and/or multispecific constructs described herein can each be prepared by any protein expression and purification method known in the art. DNA sequences encoding anti-IL-13 antibody constructs and/or multispecific constructs can be fully synthesized. Once such sequences are obtained, they are cloned into a suitable expression vector and subsequently transfected into a suitable host cell. The transfected host cells are cultured, and the supernatant is harvested and purified to obtain the anti-IL-13 antibody construct or multispecific construct described herein. In some embodiments, the host cells are lysed to obtain the expressed multispecific construct or isolated anti-TSLP antibody construct.

在一些實施例中，本申請案提供分離的核酸，其編碼本文所述之抗IL-13抗體構築體或多特異性構築體之任一者的一或多種多肽。分離的核酸可為DNA或RNA。In some embodiments, the present application provides an isolated nucleic acid encoding one or more polypeptides of any of the anti-IL-13 antibody constructs or multispecific constructs described herein. The isolated nucleic acid can be DNA or RNA.

在一些實施例中，分離的核酸被插入載體中，諸如表現載體、病毒載體或選殖載體。因此，亦提供含有本文所述之任何分離核酸的載體。為了表現核酸，可將載體導入宿主細胞中以允許核酸在宿主細胞內表現。表現載體可含有用於控制表現的多種元件，包括但不限於啟動子序列、轉錄起始序列、增強子序列、選擇標記及訊號序列。本領域之具有通常知識者可適當地選擇此等元件。舉例而言，可選擇啟動子序列以促進載體中多核苷酸的轉錄。合適的啟動子序列包括但不限於：T7啟動子、T3啟動子、SP6啟動子、β-肌動蛋白啟動子、EF1a啟動子、CMV啟動子及SV40啟動子。可選擇增強子序列以增強核酸的轉錄。可選擇可擇的標記以允許從缺乏載體的彼等細胞中選擇插入載體的宿主細胞，例如，可選擇的標記可為賦予抗生素抗性的基因。可選擇訊號序列以允許所表現的多肽被運輸至宿主細胞外部。In some embodiments, the isolated nucleic acid is inserted into a vector, such as an expression vector, a viral vector, or a selection vector. Therefore, a vector containing any of the isolated nucleic acids described herein is also provided. In order to express the nucleic acid, the vector can be introduced into a host cell to allow the nucleic acid to be expressed in the host cell. The expression vector may contain a variety of elements for controlling expression, including but not limited to a promoter sequence, a transcription initiation sequence, an enhancer sequence, a selection marker, and a signal sequence. Those of ordinary skill in the art can appropriately select these elements. For example, a promoter sequence can be selected to promote transcription of the polynucleotide in the vector. Suitable promoter sequences include, but are not limited to, the T7 promoter, the T3 promoter, the SP6 promoter, the β-actin promoter, the EF1a promoter, the CMV promoter, and the SV40 promoter. Enhancer sequences may be selected to enhance transcription of the nucleic acid. Selectable markers may be selected to allow selection of host cells harboring the vector from those lacking the vector. For example, the selectable marker may be a gene that confers antibiotic resistance. Signal sequences may be selected to allow the expressed polypeptide to be exported from the host cell.

在一些實施例中，提供一種分離的宿主細胞，其含有編碼本文所述之抗IL-13抗體構築體或多特異性構築體之任一者之多肽部分的任何分離的核酸或載體。在一些實施例中，提供一種分離的宿主細胞，其表現本文所述之抗IL-13抗體構築體或多特異性構築體之任一者。在一些實施例中，本文所述之抗IL-13抗體構築體(例如，全長抗體或Fab片段)或多特異性構築體之任一者的二或多種多肽由單一載體編碼。在一些實施例中，本文所述之抗IL-13抗體構築體(例如，全長抗體或Fab片段)或多特異性構築體之任一者的二或多種多肽由二或多種載體編碼。含有載體的宿主細胞可用於表現或選殖分離的核酸。合適的宿主細胞可包括但不限於：原核細胞、真菌細胞、酵母細胞或高等真核細胞，諸如哺乳動物細胞。抗體及抗原結合片段在原核細胞(諸如大腸桿菌)中的表現為本領域中充分證實的。相關回顧文獻，參見例如Pluckthun, A. BioTechnology 9: 545-551 (1991)。本領域具有通常知識者亦可使用真核細胞培養表現來作為生產抗體或其抗原結合片段的選項，參見回顧文獻，例如Ref, M. E. (1993) Curr. Opinion Biotech. 4: 573-576；Trill J. J.等人(1995) Curr. Opinion Biotech 6: 553-560。高等真核細胞，特別是源自多細胞生物的細胞可用於表現醣基化多肽。合適的高等真核細胞包括但不限於無脊椎動物細胞與昆蟲細胞以及脊椎動物細胞。在一些實施例中，該宿主細胞為大腸桿菌。在一些實施例中，該宿主細胞為中國倉鼠卵巢(CHO)細胞或HEK293細胞。In some embodiments, an isolated host cell is provided that contains any isolated nucleic acid or vector encoding a polypeptide portion of any of the anti-IL-13 antibody constructs or multispecific constructs described herein. In some embodiments, an isolated host cell is provided that expresses any of the anti-IL-13 antibody constructs or multispecific constructs described herein. In some embodiments, two or more polypeptides of any of the anti-IL-13 antibody constructs (e.g., full-length antibodies or Fab fragments) or multispecific constructs described herein are encoded by a single vector. In some embodiments, two or more polypeptides of any of the anti-IL-13 antibody constructs (e.g., full-length antibodies or Fab fragments) or multispecific constructs described herein are encoded by two or more vectors. Host cells containing the vector can be used to express or clone the isolated nucleic acid. Suitable host cells include, but are not limited to, prokaryotic cells, fungal cells, yeast cells, or higher eukaryotic cells, such as mammalian cells. The expression of antibodies and antigen-binding fragments in prokaryotic cells (such as Escherichia coli) is well established in the art. For a review of this literature, see, for example, Pluckthun, A. BioTechnology 9: 545-551 (1991). Those skilled in the art will also appreciate that eukaryotic cell culture expression is an option for producing antibodies or antigen-binding fragments thereof. For example, see review articles such as Ref, M.E. (1993) Curr. Opinion Biotech. 4: 573-576; Trill J.J. et al. (1995) Curr. Opinion Biotech 6: 553-560. Higher eukaryotic cells, particularly those derived from multicellular organisms, can be used to express glycosylated polypeptides. Suitable higher eukaryotic cells include, but are not limited to, invertebrate and insect cells, as well as vertebrate cells. In some embodiments, the host cell is Escherichia coli. In some embodiments, the host cell is a Chinese hamster ovary (CHO) cell or a HEK293 cell.

可使用本領域已知之任何合適方法將載體導入宿主細胞，包括但不限於：DEAE-葡聚醣媒介的遞輸、磷酸鈣沉澱法、陽離子脂質媒介的遞輸、微質體媒介的轉染、電穿孔、微粒轟擊、受體媒介的基因遞輸、由聚離胺酸、組蛋白、殼聚醣及肽媒介的遞輸。用於轉染及轉形細胞以表現感興趣載體的標準方法為本領域熟習的。在一些實施例中，該宿主細胞含有各自編碼本文所述之抗IL-13抗體構築體或多特異性構築體之任一者之多肽的二或多種載體。可按相同比率或不同比率將二或多種載體導入宿主細胞中。在一些實施例中，該宿主細胞包含含有分離核酸的單一載體，該載體編碼本文所述之抗IL-13抗體構築體或多特異性構築體之任一者的二或多種多肽。編碼多特異性構築體或分離的抗IL-13抗體構築體之二或多種多肽的二或多種核酸可處於相同啟動子控制下或處於不同啟動子控制下。舉例而言，相同啟動子控制下的二或多種核酸可經由IRES序列或編碼自裂解肽(例如，P2A、T2A)的序列連接。Vectors can be introduced into host cells using any suitable method known in the art, including, but not limited to, DEAE-dextran-mediated delivery, calcium phosphate precipitation, cationic lipid-mediated delivery, microplasm-mediated transfection, electroporation, microparticle bombardment, receptor-mediated gene delivery, and delivery mediated by polylysine, histones, chitosan, and peptides. Standard methods for transfecting and transforming cells to express vectors of interest are well known in the art. In some embodiments, the host cells contain two or more vectors each encoding a polypeptide of any of the anti-IL-13 antibody constructs or multispecific constructs described herein. The two or more vectors can be introduced into the host cells in the same ratio or in different ratios. In some embodiments, the host cell contains a single vector containing isolated nucleic acids encoding two or more polypeptides of any of the anti-IL-13 antibody constructs or multispecific constructs described herein. The two or more nucleic acids encoding the two or more polypeptides of the multispecific construct or isolated anti-IL-13 antibody construct can be under the control of the same promoter or different promoters. For example, the two or more nucleic acids under the control of the same promoter can be linked via an IRES sequence or a sequence encoding a self-cleaving peptide (e.g., P2A, T2A).

在一些實施例中，本申請案提供製造本文所述之抗IL-13抗體構築體或多特異性構築體之任一者的方法，其包含i)在適合表現抗IL-13抗體構築體或多特異性構築體的條件下培養含有本文所述之分離的核酸之任一者或本文所述之載體之任一者的分離的宿主細胞，或本文所述之分離的宿主細胞之任一者(例如，含有本文所述之分離的核酸或載體之任一者的宿主細胞)，以及ii)從該宿主細胞(例如，從細胞培養物或藉由溶解宿主細胞)獲得表現的抗IL-13抗體構築體或多特異性構築體。該分離的宿主細胞在允許插入載體之分離的核酸表現的條件下培養。適合多核苷酸表現的條件可包括但不限於：合適的培養基、培養基中合適的宿主細胞密度、必需營養素的存在、補充因子的存在、合適的溫度與濕度及不存在微生物污染物。本領域之具有通常知識者可根據表現目的適當地選擇合適的條件。在一些實施例中，該製造方法進一步包含純化獲得的抗IL-13抗體構築體或多特異性構築體之任一者。In some embodiments, the present application provides a method of making any of the anti-IL-13 antibody constructs or multispecific constructs described herein, comprising i) culturing an isolated host cell containing any of the isolated nucleic acids described herein or any of the vectors described herein, or any of the isolated host cells described herein (e.g., a host cell containing any of the isolated nucleic acids or vectors described herein) under conditions suitable for expression of the anti-IL-13 antibody construct or multispecific construct, and ii) obtaining the expressed anti-IL-13 antibody construct or multispecific construct from the host cell (e.g., from a cell culture or by lysing the host cells). The isolated host cells are cultured under conditions that allow expression of the isolated nucleic acid inserted into the vector. Suitable conditions for polynucleotide expression may include, but are not limited to, a suitable culture medium, a suitable host cell density in the culture medium, the presence of essential nutrients, the presence of supplementary factors, suitable temperature and humidity, and the absence of microbial contaminants. Those skilled in the art can appropriately select appropriate conditions based on the intended expression. In some embodiments, the production method further comprises purifying the obtained anti-IL-13 antibody construct or multispecific construct.

在一些實施例中，由宿主細胞表現的多肽可組裝在一起(例如，形成多肽複合體，諸如二聚體)並產生本文所述之抗IL-13抗體構築體或多特異性構築體之任一者 。在一些實施例中，該多肽複合體可形成在宿主細胞內部。舉例而言，該多肽複合體可藉助相關酵素及/或輔因子而形成在宿主細胞內部。在一些實施例中，該多肽複合體可從細胞中分泌出來。在一些實施例中，單獨的多肽可從宿主細胞中分泌出來，隨後在宿主細胞外部形成多肽複合體(諸如本文所述之抗IL-13抗體構築體或多特異性構築體之任一者 )。In some embodiments, polypeptides expressed by host cells can be assembled together (e.g., to form a polypeptide complex, such as a dimer) to produce any of the anti-IL-13 antibody constructs or multispecific constructs described herein. In some embodiments, the polypeptide complex can be formed inside the host cell. For example, the polypeptide complex can be formed inside the host cell with the help of relevant enzymes and/or cofactors. In some embodiments, the polypeptide complex can be secreted from the cell. In some embodiments, individual polypeptides can be secreted from the host cell and subsequently form a polypeptide complex (such as any of the anti-IL-13 antibody constructs or multispecific constructs described herein) outside the host cell.

在一些實施例中，本文所述之抗IL-13抗體構築體或多特異性構築體之任一者的二或多種多肽可單獨表現，並允許在合適的條件下形成(例如，二聚化)抗IL-13抗體構築體或多特異性構築體。舉例而言，可在合適的緩衝液中將第一多肽與第二多肽組合，以使第一蛋白質單體與第二蛋白質單體經由適當的相互作用(諸如疏水性相互作用)進行二聚化。在一些實施例中，可在含有酵素及/或輔因子的合適緩衝液中將第一多肽與第二多肽組合，該酵素及/或輔因子可促進第一多肽與第二多肽的二聚化。在一些實施例中，可在合適的媒劑中將第一多肽與第二多肽組合，以使其等在合適的試劑及/或催化劑存在下彼此反應。In some embodiments, two or more polypeptides of any of the anti-IL-13 antibody constructs or multispecific constructs described herein can be expressed separately and allowed to form (e.g., dimerize) the anti-IL-13 antibody construct or multispecific construct under suitable conditions. For example, a first polypeptide and a second polypeptide can be combined in a suitable buffer to allow dimerization of the first and second protein monomers via suitable interactions (e.g., hydrophobic interactions). In some embodiments, the first and second polypeptides can be combined in a suitable buffer containing an enzyme and/or cofactor that promotes dimerization of the first and second polypeptides. In some embodiments, the first polypeptide and the second polypeptide can be combined in a suitable medium to react with each other in the presence of a suitable reagent and/or catalyst.

可使用任何合適的方法收集表現的多肽及/或多肽複合體。多肽及/或多肽複合體可表現在細胞內、週質空間中，或分泌至細胞外的培養基中。若多肽及/或多肽複合體表現在細胞內，則可將含有多肽及/或多肽複合體的宿主細胞溶解，並藉由離心或超濾而移除不需要的碎片，從溶解物中分離多肽及/或多肽複合體。若多肽及/或多肽複合體分泌至大腸桿菌的週質空間中，則可在諸如乙酸鈉(pH 3.5)、EDTA及苯甲基磺醯基氟(PMSF)等試劑存在下將細胞漿糊解凍約30分鐘，並可藉由離心移除細胞碎片(Carter等人，BioTechnology 10:163-167 (1992))。若多肽及/或多肽複合體分泌至培養基中，則可收集細胞培養物的上清液，並使用市售蛋白質濃縮過濾器(例如，Amincon或Millipore Pellicon超濾裝置)進行濃縮。可在收集及濃縮步驟中添加蛋白酶抑制劑及/或抗生素，以抑制蛋白質降解及/或污染性微生物的生長。The expressed polypeptide and/or polypeptide complex can be collected using any suitable method. The polypeptide and/or polypeptide complex can be expressed intracellularly, in the periplasmic space, or secreted into the culture medium outside the cell. If the polypeptide and/or polypeptide complex is expressed intracellularly, the host cells containing the polypeptide and/or polypeptide complex can be lysed and the polypeptide and/or polypeptide complex can be isolated from the lysate by centrifugation or ultrafiltration to remove unwanted debris. If the polypeptide and/or polypeptide complex is secreted into the periplasmic space of E. coli, the cell slurry can be thawed in the presence of reagents such as sodium acetate (pH 3.5), EDTA, and phenylmethylsulfonyl fluoride (PMSF) for about 30 minutes, and cell debris can be removed by centrifugation (Carter et al., BioTechnology 10:163-167 (1992)). If the polypeptide and/or polypeptide complex is secreted into the culture medium, the supernatant of the cell culture can be collected and concentrated using commercially available protein concentrator filters (e.g., Amincon or Millipore Pellicon ultrafiltration devices). Protease inhibitors and/or antibiotics may be added during the collection and concentration steps to inhibit protein degradation and/or the growth of contaminating microorganisms.

可藉由合適的方法進一步純化所表現的多肽及/或多肽複合體，諸如但不限於：親和力層析法、羥基磷灰石層析法、尺寸排阻層析法、凝膠電泳、透析、離子交換管柱上的離子交換分離等、乙醇沉澱、逆相HPLC、矽膠層析法、肝素瓊脂糖層析法、陰離子或陽離子交換樹脂層析法(諸如，聚天門冬胺酸管柱)、聚焦層析法、SDS-PAGE及硫酸銨沉澱(回顧文獻，參見Bonner, P. L., Protein purification，由Taylor & Francis出版，2007；Janson, J. C.,等人，Protein purification: principles, high resolution methods and applications，由Wiley-VCH出版，1998)。The expressed polypeptide and/or polypeptide complex can be further purified by suitable methods, such as, but not limited to, affinity chromatography, hydroxyapatite chromatography, size exclusion chromatography, gel electrophoresis, dialysis, ion exchange separation on an ion exchange column, ethanol precipitation, reversed-phase HPLC, silica gel chromatography, heparin-agarose chromatography, anion or cation exchange resin chromatography (e.g., polyaspartic acid column), focusing chromatography, SDS-PAGE, and ammonium sulfate precipitation (for review, see Bonner, P. L., Protein purification, published by Taylor & Francis, 2007; Janson, J. C., et al., Protein purification: principles, High Resolution Methods and Applications, published by Wiley-VCH, 1998).

在一些實施例中，可藉由親和力層析法來純化多肽及/或多肽複合體。在一些實施例中，蛋白A層析法或蛋白A/G (蛋白A與蛋白G的融合蛋白)層析法可用於純化含有源自抗體CH2域及/或CH3域組分的多肽及/或多肽複合體(Lindmark等人，J. Immunol. Meth. 62:1-13 (1983))；Zettlit, K. A., Protein Engineering，第V部分，531-535, 2010)。在一些實施例中，蛋白G層析法可用於純化含有IgG γ3重鏈的多肽及/或多肽複合體(Guss等人，EMBO J. 5:1567 1575 (1986))。在一些實施例中，蛋白L層析法可用於純化含有κ 輕鏈的多肽及/或多肽複合體(Sudhir, P.，Antigen engineering protocols，第26章，由Humana Press出版，1995；Nilson, B. H. K.等人，J. Biol. Chem., 267, 2234-2239 (1992))。親和配體附接的基質通常為瓊脂糖，但亦可使用其他基質。機械穩定的基質(諸如可控孔徑玻璃或聚(苯乙烯二乙烯基)苯)允許達成比瓊脂糖更快的流速及更短的處理時間。當任何抗IL-13抗體構建體包含額外的CH3域時，可使用Bakerbond ABX樹脂(J. T. Baker，Phillipsburg，N.J.)進行純化。IV.醫藥組成物、單位劑量、製品及套組In some embodiments, polypeptides and/or polypeptide complexes can be purified by affinity chromatography. In some embodiments, protein A chromatography or protein A/G (a fusion protein of protein A and protein G) chromatography can be used to purify polypeptides and/or polypeptide complexes containing components derived from antibody CH2 and/or CH3 domains (Lindmark et al., J. Immunol. Meth. 62:1-13 (1983)); Zettlit, KA, Protein Engineering, Part V, 531-535, 2010). In some embodiments, protein G chromatography can be used to purify polypeptides and/or polypeptide complexes containing IgG γ3 heavy chains (Guss et al., EMBO J. 5:1567-1575 (1986)). In some embodiments, protein L chromatography can be used to purify polypeptides and/or polypeptide complexes containing kappa light chains (Sudhir, P., Antigen Engineering Protocols, Chapter 26, Humana Press, 1995; Nilson, BHK et al., J. Biol. Chem., 267, 2234-2239 (1992)). The matrix to which the affinity ligand is attached is typically agarose, but other matrices can also be used. Mechanically stable matrices (such as controlled pore glass or poly(styrenedivinyl)benzene) allow for faster flow rates and shorter processing times than agarose. When any anti-IL-13 antibody construct contains an additional CH3 domain, Bakerbond ABX resin (JT Baker, Phillipsburg, NJ) can be used for purification.IV.Pharmaceutical compositions, unit doses, products, and kits

本申請案進一步提供含有本發明之抗IL-13抗體構築體或多特異性構築體及可選地醫藥上可接受之載體的醫藥組成物。This application further provides a pharmaceutical composition comprising the anti-IL-13 antibody construct or multispecific construct of the present invention and, optionally, a pharmaceutically acceptable carrier.

本文所述之抗IL-13抗體構築體或多特異性構築體或其醫藥組成物可適合本文所述之多種投予方式，包括例如全身或局部投予。在一些實施例中，該醫藥組成物係調配成靜脈內投予。在一些實施例中，該醫藥組成物係調配成皮下注射。The anti-IL-13 antibody constructs or multispecific constructs described herein, or pharmaceutical compositions thereof, can be suitable for a variety of administration routes as described herein, including, for example, systemic or local administration. In some embodiments, the pharmaceutical composition is formulated for intravenous administration. In some embodiments, the pharmaceutical composition is formulated for subcutaneous injection.

如本文所用，「載劑」包括醫藥上可接受之載體賦形劑或穩定劑，其在所用劑量及濃度下對暴露於其之細胞或哺乳動物無毒。通常生理上可接受之載劑為pH緩衝水溶液。可接受之載劑、賦形劑或穩定劑在所用劑量及濃度下對接受者無毒，且包括：緩衝液，諸如磷酸鹽、檸檬酸鹽及其他有機酸；抗氧化劑，包括抗壞血酸及甲硫胺酸；防腐劑(諸如十八烷基二甲基苄基氯化銨；六甲氯銨；氯化苄烷銨；氯化苄甲乙氧銨；酚類、丁醇或苄醇；對羥基苯甲酸烷基酯，諸如對羥基苯甲酸甲酯或對羥基苯甲酸丙酯；兒茶酚；間苯二酚；環己醇；3-戊醇；及間甲酚)；低分子量(小於約10個殘基)多肽；蛋白質，諸如血清白蛋白、明膠或免疫球蛋白；親水聚合物，諸如聚乙烯吡咯啶酮；胺基酸，諸如甘胺酸、麩醯胺酸、天冬醯胺酸、組胺酸、精胺酸或離胺酸；單醣、雙醣及其他碳水化合物，包括葡萄糖、甘露糖或糊精；螯合劑，諸如EDTA；糖，諸如蔗糖、甘露醇、海藻糖或山梨醇；鹽形成反離子，諸如鈉；金屬複合體(例如Zn-蛋白質複合體)；及/或非離子型表面活性劑，諸如TWEEN™、PLURONICS™或聚乙二醇(PEG)。As used herein, "carrier" includes a pharmaceutically acceptable carrier form or stabilizer that is nontoxic to cells or mammals exposed thereto at the dosages and concentrations employed. Typically, a physiologically acceptable carrier is a pH-buffered aqueous solution. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed and include: buffers such as phosphates, citrates, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzylammonium chloride; hexamethylammonium chloride; benzylammonium chloride; benzylmethylethoxyammonium chloride; phenols, butyl or benzyl alcohol; alkyl parahydroxybenzoates such as methyl or propyl parahydroxybenzoate; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, Such as serum albumin, gelatin, or immunoglobulin; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, aspartic acid, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrin; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose, or sorbitol; salt-forming counterions such as sodium; metal complexes (e.g., Zn-protein complexes); and/or non-ionic surfactants such as TWEEN™, PLURONICS™, or polyethylene glycol (PEG).

在一些實施例中，該醫藥組成物係調配成pH範圍為約4.5至約9.0，包括例如pH範圍為約5.0至約8.0、約6.5至約7.5、約6.5至約7.0或約7.0至約7.5之任一者。在一些實施例中，該醫藥組成物亦可藉由添加合適的張力調節劑(諸如甘油)而使其與血液等張。In some embodiments, the pharmaceutical composition is formulated to have a pH range of about 4.5 to about 9.0, including, for example, a pH range of about 5.0 to about 8.0, about 6.5 to about 7.5, about 6.5 to about 7.0, or about 7.0 to about 7.5. In some embodiments, the pharmaceutical composition can also be made isotonic with blood by adding a suitable tonicity adjusting agent (such as glycerol).

用於體內投予的醫藥組成物通常調配成無菌的、實質上等張的且完全符合美國食品藥品管理局的所有藥品優良製造規範(GMP)規定。經由無菌濾膜過濾即可輕易實現無菌。在一些實施例中，該組成物不含任何病原體。為了注射，該醫藥組成物可呈水溶液形式，例如生理學上相容之緩衝液(諸如漢克氏溶液(Hank's solution)或林格氏溶液(Ringer's solution))。此外，該醫藥組成物可呈固體形式，並在使用前立即重新溶解或懸浮。亦包括冷凍乾燥組成物。Pharmaceutical compositions for intravenous administration are typically formulated to be sterile, substantially isotonic, and in full compliance with all Good Manufacturing Practice (GMP) regulations of the U.S. Food and Drug Administration. Sterility is readily achieved by filtration through a sterile filter membrane. In some embodiments, the composition is free of pathogens. For injection, the pharmaceutical composition may be in the form of an aqueous solution, such as a physiologically compatible buffer (such as Hank's solution or Ringer's solution). Alternatively, the pharmaceutical composition may be in solid form and reconstituted or suspended immediately prior to use. Freeze-dried compositions are also included.

在一些實施例中，該醫藥組成物依照常規程序調配為適應於非經口(例如，靜脈內、肌內或皮下)投予的醫藥組成物。通常，注射用組成物為溶於無菌等張緩衝水溶液中的溶液。當必要時，該組成物亦可包括增溶劑及局部麻醉劑(諸如利多卡因(lidocaine))，以減輕注射部位的疼痛。一般而言，各成分可單獨提供或以單位劑量形式混合在一起，例如以凍乾粉末或無水濃縮物形式置於密封容器(諸如安瓿瓶或小袋)中，並標註活性劑的量。當藉由輸注來投予組成物時，可使用含有無菌醫藥級水或鹽水的輸液瓶進行分配。當藉由注射而投予組成物時，可提供無菌注射用水或鹽水的安瓿瓶，以便在投予前將各成分混合。In some embodiments, the pharmaceutical composition is formulated as a pharmaceutical composition suitable for non-oral (e.g., intravenous, intramuscular or subcutaneous) administration according to conventional procedures. Typically, the composition for injection is a solution dissolved in a sterile isotonic buffered aqueous solution. When necessary, the composition may also include a solubilizer and a local anesthetic (such as lidocaine) to reduce pain at the injection site. Generally speaking, the components can be provided separately or mixed together in unit dose form, for example, in the form of a lyophilized powder or anhydrous concentrate in a sealed container (such as an ampoule or a sachet) and labeled with the amount of active agent. When the composition is administered by infusion, it can be dispensed using an infusion bottle containing sterile pharmaceutical grade water or saline. When the composition is administered by injection, an ampoule of sterile water for injection or saline is provided so that the ingredients can be mixed prior to administration.

在一些實施例中，適合向哺乳動物(諸如人類)投予醫藥組成物。在一些實施例中，適合向囓齒類動物(例如，小鼠、大鼠)或非人類靈長類動物(例如，食蟹獼猴)投予醫藥組成物。在一些實施例中，該醫藥組成物係含於一次性小瓶(諸如一次性密封小瓶)中。在一些實施例中，該醫藥組成物係含於多次性小瓶中。在一些實施例中，該醫藥組成物以散裝形式裝在容器中。在一些實施例中，該醫藥組成物係冷凍保存。In some embodiments, the pharmaceutical composition is suitable for administration to mammals (e.g., humans). In some embodiments, the pharmaceutical composition is suitable for administration to rodents (e.g., mice, rats) or non-human primates (e.g., cynomolgus macaques). In some embodiments, the pharmaceutical composition is contained in a disposable vial (e.g., a sealed disposable vial). In some embodiments, the pharmaceutical composition is contained in a multi-use vial. In some embodiments, the pharmaceutical composition is packaged in bulk in a container. In some embodiments, the pharmaceutical composition is stored frozen.

亦提供本文所述之抗IL-13抗體構築體或多特異性構築體或其組成物(諸如醫藥組成物)之任一者的單位劑量形式。術語「單位劑量形式」意指適合作為個體(例如，人類)單位劑量的物理上離散單位，每個單位含有預定量的活性物質，經計算以產生所需治療效果，並結合合適的醫藥載劑、稀釋劑或賦形劑。此等單位劑量形式可以單一或多個單位劑量儲存在合適的包裝中，亦可進一步滅菌及密封。Also provided are unit dosage forms of any of the anti-IL-13 antibody constructs or multispecific constructs described herein, or compositions thereof (e.g., pharmaceutical compositions). The term "unit dosage form" refers to physically discrete units suitable as individual (e.g., human) unit doses, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical carrier, diluent, or formulation. Such unit dosage forms can be stored as single or multiple unit doses in suitable packaging, which may be further sterilized and sealed.

本申請案進一步提供含有適用於包裝的本文所述之組成物(諸如醫藥組成物)的製品。本文所述之組成物(諸如醫藥組成物)的適用包裝為本領域已知的，包括例如小瓶(諸如密封小瓶)、容器、安瓿瓶、瓶子、罐子、撓性包裝(例如，密封聚酯樹脂(Mylar)或塑料袋)及其類似物。此等製品可進一步滅菌及/或密封。This application further provides articles of manufacture containing the compositions described herein (e.g., pharmaceutical compositions) suitable for packaging. Suitable packaging for the compositions described herein (e.g., pharmaceutical compositions) is known in the art and includes, for example, vials (e.g., sealed vials), containers, ampoules, bottles, jars, flexible packaging (e.g., sealed Mylar or plastic bags), and the like. Such articles of manufacture may be further sterilized and/or sealed.

本申請案亦提供含有本文所述之組成物(諸如醫藥組成物)的套組，並可進一步包含使用該組成物之方法(諸如本文所述之用途)的說明書。本文所述之套組可進一步包括從商業及用戶觀點而言所需的其他材料，包括其他緩衝液、稀釋劑、過濾器、針頭、注射器及/或塗藥器以及帶有用於進行本文所述之任何方法的說明書的包裝仿單。VI.治療發炎性疾病的方法This application also provides kits containing the compositions described herein (such as pharmaceutical compositions) and may further include instructions for methods of using the compositions (such as the uses described herein). The kits described herein may further include other materials necessary from a commercial and user perspective, including other buffers, diluents, filters, needles, syringes and/or applicators, and packaging instructions for performing any of the methods described herein.VI.Methods of Treating Inflammatory Diseases

亦提供治療個體(例如，人類)發炎性疾病的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之本文所述之多特異性構築體、分離的抗IL-13抗體構築體或醫藥組成物之任一者。在一些實施例中，該發炎性疾病為氣喘(例如，嚴重氣喘)、異位性皮膚炎(例如，中度至嚴重異位性皮膚炎)或慢性阻塞性肺病(COPD)。在一些實施例中，該方法進一步包含同時或依序投予有效量之皮質類固醇與該多特異性構築體、分離的抗IL-13抗體構築體或其醫藥組成物。Also provided are methods for treating an inflammatory disease in a subject (e.g., a human) comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of any of the multispecific constructs, isolated anti-IL-13 antibody constructs, or pharmaceutical compositions described herein. In some embodiments, the inflammatory disease is asthma (e.g., severe asthma), atopic dermatitis (e.g., moderate to severe atopic dermatitis), or chronic obstructive pulmonary disease (COPD). In some embodiments, the method further comprises administering an effective amount of a corticosteroid concurrently or sequentially with the multispecific construct, isolated anti-IL-13 antibody construct, or pharmaceutical composition thereof.

在一些實施例中，該發炎性疾病為IL-13相關疾病。與IL-13相關的發炎性疾病可為由IL-13之表現引起的或與其相關的疾病，例如與健康狀態相比過度表現，或在不同於健康狀態的位置處(例如，在不同的細胞或組織上)錯誤表現。此類發炎性疾病可進一步與一或多種抗原(例如，TSLP)相關，其中該發炎性疾病進一步由一或多種此等抗原(例如，TSLP)之表現引起或與其相關，例如與健康狀態相比過度表現，或在不同於健康狀態的位置處錯誤表現。In some embodiments, the inflammatory disease is an IL-13-associated disease. An IL-13-associated inflammatory disease can be caused by or associated with the expression of IL-13, such as overexpression compared to a healthy state or misexpression at a different location (e.g., on different cells or tissues) than a healthy state. Such inflammatory diseases can further be associated with one or more antigens (e.g., TSLP), wherein the inflammatory disease is further caused by or associated with the expression of one or more such antigens (e.g., TSLP), such as overexpression compared to a healthy state or misexpression at a different location than a healthy state.

可藉由本文所述之任何治療方法治療的發炎性疾病之實例包括但不限於：異位性皮膚炎、氣喘、過敏性鼻炎(allergic rhinitis)、嗜酸性細胞性食道炎(eosinophilic esophagitis)、類風濕性關節炎(rheumatoid arthritis)、乾癬(psoriasis)、慢性阻塞性肺病(COPD)及乳糜瀉(celiac disease)。在一些實施例中，該發炎性疾病為自體免疫疾病或自體免疫病狀，諸如異位性皮膚炎、狼瘡(lupus)、休格連氏症候群(Sjogren's syndrome)、多發性硬化症(multiple sclerosis)、乾癬、乾癬性關節炎(psoriatic arthritis)、類風濕性關節炎、類肉瘤病(sarcoidosis)、潰瘍性結腸炎(ulcerative colitis)等。在一些實施例中，該發炎性疾病為另一種與免疫功能有關的疾病 (例如，氣喘、子宮內膜異位症(endometriosis)、纖維化(fibrosis))。在一些實施例中，該發炎性疾病過敏反應，諸如氣喘、過敏性鼻炎、嗜酸性細胞性食道炎等。在一些實施例中，該發炎性疾病為纖維化發炎性腸道疾病。在一些實施例中，該發炎性疾病為氣喘或異位性皮膚炎，諸如任何嚴重程度的氣喘或異位性皮膚炎。Examples of inflammatory diseases that may be treated by any of the treatment methods described herein include, but are not limited to, atopic dermatitis, asthma, allergic rhinitis, eosinophilic esophagitis, rheumatoid arthritis, psoriasis, chronic obstructive pulmonary disease (COPD), and celiac disease. In some embodiments, the inflammatory disease is an autoimmune disease or autoimmune condition, such as atopic dermatitis, lupus, Sjögren's syndrome, multiple sclerosis, pityriasis, psoriasis arthritis, rheumatoid arthritis, sarcoidosis, ulcerative colitis, etc. In some embodiments, the inflammatory disease is another disease related to immune function (e.g., asthma, endometriosis, fibrosis). In some embodiments, the inflammatory disease is an allergic reaction, such as asthma, allergic rhinitis, eosinophilic esophagitis, etc. In some embodiments, the inflammatory disease is fibrosing inflammatory bowel disease. In some embodiments, the inflammatory disease is asthma or atopic dermatitis, such as asthma or atopic dermatitis of any severity.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：第一抗體部分，其為scFv (「抗IL-13 scFv」)，以及第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的全長抗體，其中該抗IL-13 scFv係經由視情況的連接子融合至第二抗體部分之重鏈之一者的C端以形成融合多肽。在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)，以及第二抗體部分，其為特異性地結合至第二標靶抗原的全長抗體(例如，TSLP；「抗TSLP全長抗體」)；其中抗IL-13 scFv1係融合至第二抗體部分之第一重鏈的C端以形成第一融合多肽，且抗IL-13 scFv2係融合至第二抗體部分之第二重鏈的C端以形成第二融合多肽。在一些實施例中，該抗TSLP全長抗體包含兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈及兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈，且該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:113之胺基酸序列。In some embodiments, a method is provided for treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or a pharmaceutical composition thereof) comprising: a first antibody portion that is a scFv ("anti-IL-13 scFv"), and a second antibody portion that is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP), wherein the anti-IL-13 scFv is fused to the C-terminus of one of the heavy chains of the second antibody portion via an optional linker to form a fusion polypeptide. In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or a pharmaceutical composition thereof) comprising: two anti-IL-13 antibody portions, each of which is a scFv ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP; "anti-TSLP full-length antibody"); wherein the anti-IL-13 scFv1 is fused to the C-terminus of the first heavy chain of the second antibody portion to form a first fusion polypeptide, and the anti-IL-13 scFv2 is fused to the C-terminus of the second heavy chain of the second antibody portion to form a second fusion polypeptide. In some embodiments, the anti-TSLP full-length antibody comprises two light chains, each comprising the amino acid sequence of SEQ ID NO: 103, and two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 105, and the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of SEQ ID NO: 113.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)、兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-抗IL-13 scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-抗IL-13 scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成Fab2。在一些實施例中，該多特異性構築體包含各自含有SEQ ID NO:103之胺基酸序列的第一多肽及第四多肽，以及各自含有SEQ ID NO:110之胺基酸序列的第二多肽及第三多肽。In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or pharmaceutical composition thereof) comprising: two anti-IL-13 antibody portions, which are scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), two second antibody portions, which are Fabs that specifically bind to a second target antigen (e.g., TSLP), ("Fab1" and "Fab2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optional linker-anti-IL-13 scFv1-optional linker-Fc domain; iii) a second polypeptide comprising N' to C': VH2-(H2-CH1)-optional linker-anti-IL-13 scFv2-optionally, a linker-a third polypeptide of the second subunit of the Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form Fab2. In some embodiments, the multispecific construct comprises the first and fourth polypeptides, each comprising the amino acid sequence of SEQ ID NO: 103, and the second and third polypeptides, each comprising the amino acid sequence of SEQ ID NO: 110.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：第一抗體部分，其為抗IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)、第二抗體部分，其為特異性地結合至第二標靶抗原(例如，TSLP)的scFv，且其中該scFv部分係經由視情況的連接子融合至抗IL-13全長抗體之重鏈之一者的C端以形成融合多肽。在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：第一抗體部分，其為抗IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的scFv (「scFv1」及「scFv2」)；且其中scFv1係經由視情況的連接子融合至抗IL-13全長抗體之第一重鏈的C端以形成第一融合多肽，且scFv2係經由視情況的連接子融合至抗IL-13全長抗體之第二重鏈的C端以形成第二融合多肽。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102之胺基酸序列的輕鏈，其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:111之胺基酸序列。在一些實施例中，該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:220-223之任一者的胺基酸序列。In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or a pharmaceutical composition thereof) comprising: a first antibody portion that is a full-length anti-IL-13 antibody (e.g., any of the full-length anti-IL-13 antibodies described herein); and a second antibody portion that is a scFv that specifically binds to a second target antigen (e.g., TSLP), wherein the scFv portion is fused to the C-terminus of one of the heavy chains of the full-length anti-IL-13 antibody via an optional linker to form a fusion polypeptide. In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or a pharmaceutical composition thereof) comprising: a first antibody portion that is a full-length anti-IL-13 antibody (e.g., any of the full-length anti-IL-13 antibodies described herein), and two second antibody portions that are scFvs that specifically bind to a second target antigen (e.g., TSLP). ("scFv1" and "scFv2"); wherein scFv1 is fused to the C-terminus of the first heavy chain of the anti-IL-13 full-length antibody via a linker, as appropriate, to form a first fusion polypeptide, and scFv2 is fused to the C-terminus of the second heavy chain of the anti-IL-13 full-length antibody via a linker, as appropriate, to form a second fusion polypeptide. In some embodiments, the anti-IL-13 full-length antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 125, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 102, wherein the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of SEQ ID NO: 111. In some embodiments, the full-length anti-IL-13 antibody comprises two heavy chains, each comprising the amino acid sequence of SEQ ID NO: 225, and two light chains, each comprising the amino acid sequence of SEQ ID NO: 102 or 197, and wherein the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of any one of SEQ ID NOs: 220-223.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)、兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的scFv (「scFv1」及「scFv2」)，以及Fc域；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-scFv1-視情況的連接子-Fc域之第一次單元的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及iv)含有N’至C’：VL2-(L2-CL)的第四多肽；且其中VH1-(H1-CH1)及VL1-(L1-CL)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2。在一些實施例中，該多特異性構築體包含各自含有SEQ ID NO:102或197之胺基酸序列的第一多肽及第四多肽，以及各自含有SEQ ID NO:112之胺基酸序列的第二多肽及第三多肽。In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or pharmaceutical composition thereof) comprising: two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2," e.g., any of the anti-IL-13 Fabs described herein); and two second antibody portions, which are scFvs that specifically bind to a second target antigen (e.g., TSLP). ("scFv1" and "scFv2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optional linker-scFv1-optional linker-Fc domain; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optional linker-scFv2-optional linker-Fc domain; and iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); and wherein VH1-(H1-CH1) and VL1-(L1-CL) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-IL-13 Fab2. In some embodiments, the multispecific construct comprises a first polypeptide and a fourth polypeptide, each comprising the amino acid sequence of SEQ ID NO: 102 or 197, and a second polypeptide and a third polypeptide, each comprising the amino acid sequence of SEQ ID NO: 112.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原的全長抗體(例如，TSLP；「抗TSLP全長抗體」)；其中該多特異性構築體包含：i)含有第二抗體部分之第一輕鏈的第一多肽；ii)含有N’至C’：第二抗體部分之第一重鏈-視情況的連接子-VH1-(H1-CH1)的第二多肽；iii)含有N’至C’：第二抗體部分之第二重鏈-視情況的連接子-VH2-(H2-CH1)的第三多肽；及iv)含有第二抗體部分之第二輕鏈的第四多肽；v)含有N’至C’：VL1-(L1-CL)的第五多肽；及vi)含有N’至C’：VL2-(L2-CL)的第六多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VL2-(L2-CL)及VH2-(H2-CH1)形成抗IL-13 Fab2。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該全長抗體之輕鏈係衍生自κ輕鏈。In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or pharmaceutical composition thereof) comprising: two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2," such as the anti-IL-13 described herein; Fab), and a second antibody portion that is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP; "anti-TSLP full-length antibody"); wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain of the second antibody portion; ii) a second polypeptide comprising the first heavy chain of the second antibody portion, optionally a linker, VH1-(H1-CH1); iii) a second polypeptide comprising the first heavy chain of the second antibody portion, optionally a linker, VH1-(H1-CH1); ': the second heavy chain of the second antibody portion - optionally a linker - VH2-(H2-CH1); and iv) a fourth polypeptide comprising the second light chain of the second antibody portion; v) a fifth polypeptide comprising N' to C': VL1-(L1-CL); and vi) a sixth polypeptide comprising N' to C': VL2-(L2-CL); wherein VL1-(L1-CL) and VH1-(H1-CH1) form anti-IL-13 Fab1, and VL2-(L2-CL) and VH2-(H2-CH1) form anti-IL-13 Fab2. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the light chain of the full-length antibody is derived from a kappa light chain.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：抗IL-13抗體部分，其為全長抗體(「抗IL-13全長抗體」；例如本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」))；其中該多特異性構築體包含：i)含有抗IL-13全長抗體之第一輕鏈(L1)的第一多肽；ii)含有N’至C’：抗IL-13全長抗體之第一重鏈(H1)-視情況的連接子-VH3-(H3-CH1)的第二多肽；iii)含有N’至C’：抗IL-13全長抗體之第二重鏈(H2)-視情況的連接子-VH4-(H4-CH1)的第三多肽；及iv)含有抗IL-13全長抗體之第二輕鏈(L2)的第四多肽；v)含有N’至C’：VL3-(L3-CL)的第五多肽；及vi)含有N’至C’：VL4-(L4-CL)的第六多肽；且其中VL3-(L3-CL)及VH3-(H3-CH1)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，L3-CL及L4-CL係衍生自κ輕鏈。In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or a pharmaceutical composition thereof) comprising: an anti-IL-13 antibody portion that is a full-length antibody ("anti-IL-13 full-length antibody"; e.g., any of the anti-IL-13 full-length antibodies described herein); and two second antibody portions that are Fabs that specifically bind to a second target antigen (e.g., TSLP). ("Fab1" and "Fab2")); wherein the multispecific construct comprises: i) a first polypeptide comprising the first light chain (L1) of an anti-IL-13 full-length antibody; ii) a second polypeptide comprising N' to C': the first heavy chain (H1) of an anti-IL-13 full-length antibody - an optional linker - VH3 - (H3-CH1); iii) a second polypeptide comprising N' to C': the second heavy chain (H2) of an anti-IL-13 full-length antibody - an optional linker - VH4 - (H4 -CH1); and iv) a fourth polypeptide containing the second light chain (L2) of the anti-IL-13 full-length antibody; v) a fifth polypeptide containing N' to C': VL3-(L3-CL); and vi) a sixth polypeptide containing N' to C': VL4-(L4-CL); and wherein VL3-(L3-CL) and VH3-(H3-CH1) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, L3-CL and L4-CL are derived from a kappa light chain.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」，例如本文所述之抗IL-13 Fab之任一者)，以及第二抗體部分，其為特異性地結合至第二標靶抗原的全長抗體(例如，TSLP；「抗TSLP全長抗體」)；其中該多特異性構築體包含：i)含有N’至C’：VL1-(L1-CL)的第一多肽；ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-第二抗體部分之第一重鏈的第二多肽；iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-第二抗體部分之第二重鏈的第三多肽；iv)含有N’至C’：VL2-(L2-CL)的第四多肽；v)含有第二抗體部分之第一輕鏈的第五多肽；及vi)含有第二抗體部分之第二輕鏈的第六多肽；且其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2。在一些實施例中，該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中：(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，該全長抗體之輕鏈係衍生自κ輕鏈。In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or pharmaceutical composition thereof) comprising: two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2," such as the anti-IL-13 described herein; Fab), and a second antibody portion that is a full-length antibody that specifically binds to a second target antigen (e.g., TSLP; an "anti-TSLP full-length antibody"); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-a first heavy chain of the second antibody portion; iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optionally a linker-the second heavy chain of the second antibody portion; iv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); v) a fifth polypeptide comprising the first light chain of the second antibody portion; and vi) a sixth polypeptide comprising the second light chain of the second antibody portion; wherein VL1-(L1-CL) and VH1-(H1-CH1) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form an anti-IL-13 Fab2. In some embodiments, the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein: (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, the light chain of the full-length antibody is derived from a kappa light chain.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：第一抗體部分，其為IL-13全長抗體(例如，本文所述之抗IL-13全長抗體之任一者)，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原(例如，TSLP)的Fab (「Fab1」及「Fab2」)；其中該多特異性構築體包含：i)含有N’至C’：VL3-(L3-CL)的第一多肽；ii)含有N’至C’：VH3-(H3-CH1)-視情況的連接子-抗IL-13全長抗體之第一重鏈(H1)的第二多肽；iii)含有N’至C’：VH4-(H4-CH1)-視情況的連接子-抗IL-13全長抗體之第二重鏈(H2)的第三多肽；iv)含有N’至C’：VL4-(L4-CL)的第四多肽；v)含有N’至C’：抗IL-13全長抗體之第一輕鏈(L1)的第五多肽；及vi)含有N’至C’：抗IL-13全長抗體之第二輕鏈(L2)的第六多肽；且其中VL3-(L3-CL)及VH3-(H3-CH1)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2。在一些實施例中，(a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代；(b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代；(c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；(d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C (或Q124C)、L135F及C214S取代；或e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C (或Q124C)及C214S取代；且其中該胺基酸位置係根據EU編號。在一些實施例中，L3-CL及L4-CL係衍生自κ輕鏈。In some embodiments, a method of treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or a pharmaceutical composition thereof) comprising: a first antibody portion that is a full-length IL-13 antibody (e.g., any of the full-length anti-IL-13 antibodies described herein), and two second antibody portions that are Fabs that specifically bind to a second target antigen (e.g., TSLP). ("Fab1" and "Fab2"); wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL3-(L3-CL); ii) a second polypeptide comprising N' to C': VH3-(H3-CH1)-optionally a linker-a first heavy chain (H1) of an anti-IL-13 full-length antibody; iii) a third polypeptide comprising N' to C': VH4-(H4-CH1)-optionally a linker-a second heavy chain (H2) of an anti-IL-13 full-length antibody; polypeptide; iv) a fourth polypeptide comprising N' to C': VL4-(L4-CL); v) a fifth polypeptide comprising N' to C': the first light chain (L1) of the anti-IL-13 full-length antibody; and vi) a sixth polypeptide comprising N' to C': the second light chain (L2) of the anti-IL-13 full-length antibody; and wherein VL3-(L3-CL) and VH3-(H3-CH1) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2. In some embodiments, (a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions; (c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; (d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C), L135F, and C214S substitutions; or e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C (or Q124C) and C214S substitutions; and wherein the amino acid positions are according to EU numbering. In some embodiments, L3-CL and L4-CL are derived from a kappa light chain.

在一些實施例中，提供一種治療個體(諸如人類)發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法，其包含向個體投予(例如，靜脈內或皮下)有效量之多特異性構築體(或其醫藥組成物)，其包含：異二聚體雙特異性抗體，其包含：i)含有VL1及L1-CL的第一輕鏈(L1)；ii)含有VH1及H1-CH1以及Fc域之第一次單元的第一重鏈(H1)；iii)含有VH2及H2-CH1以及Fc域之第二次單元的第二重鏈(H2)；及iv)含有VL2及L2-CL的第二輕鏈(L2)；其中H1及L1形成抗IL-13抗體部分，且H2及L2形成特異性地結合至第二標靶抗原(例如，TSLP)的第二抗體部分。在一些實施例中，該多特異性構築體包含：(i)含有SEQ ID NO:139之胺基酸序列的H1、含有SEQ ID NO:122或207之胺基酸序列的L1、含有SEQ ID NO:136之胺基酸序列的H2及含有SEQ ID NO:103之胺基酸序列的L2；(ii)含有SEQ ID NO:138之胺基酸序列的H1、含有SEQ ID NO:122或207之胺基酸序列的L1、含有SEQ ID NO:137之胺基酸序列的H2及含有SEQ ID NO:103之胺基酸序列的L2；(iii)含有SEQ ID NO:141之胺基酸序列的H1、含有SEQ ID NO:122或207之胺基酸序列的L1、含有SEQ ID NO:136之胺基酸序列的H2及含有SEQ ID NO:103之胺基酸序列的L2；(vi)含有SEQ ID NO:140之胺基酸序列的H1、含有SEQ ID NO:122或207之胺基酸序列的L1、含有SEQ ID NO:137之胺基酸序列的H2及含有SEQ ID NO:103之胺基酸序列的L2；(v)含有SEQ ID NO:126之胺基酸序列的H1、含有SEQ ID NO:120或208之胺基酸序列的L1、含有SEQ ID NO:136之胺基酸序列的H2及含有SEQ ID NO:103之胺基酸序列的L2；(vi)含有SEQ ID NO:127之胺基酸序列的H1、含有SEQ ID NO:120或208之胺基酸序列的L1、含有SEQ ID NO:137之胺基酸序列的H2及含有SEQ ID NO:103之胺基酸序列的L2；(vii)含有SEQ ID NO:128之胺基酸序列的H1、含有SEQ ID NO:120或208之胺基酸序列的L1、含有SEQ ID NO:136之胺基酸序列的H2及含有SEQ ID NO:103之胺基酸序列的L2；或(viii)含有SEQ ID NO:129之胺基酸序列的H1、含有SEQ ID NO:120或208之胺基酸序列的L1、含有SEQ ID NO:137之胺基酸序列的H2及含有SEQ ID NO:103之胺基酸序列的L2。In some embodiments, a method for treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) in a subject (e.g., a human) is provided, comprising administering to the subject (e.g., intravenously or subcutaneously) an effective amount of a multispecific construct (or a pharmaceutical composition thereof) comprising: a heterodimeric bispecific antibody comprising: i) a first light chain (L1) comprising VL1 and L1-CL; ii) a first light chain (L2) comprising VH1 and H1-CL; The invention relates to a first heavy chain (H1) of a first subunit comprising a CH1 and an Fc domain; iii) a second heavy chain (H2) of a second subunit comprising a VH2 and H2-CH1 and an Fc domain; and iv) a second light chain (L2) comprising VL2 and L2-CL; wherein H1 and L1 form the anti-IL-13 antibody portion, and H2 and L2 form the second antibody portion that specifically binds to a second target antigen (e.g., TSLP). In some embodiments, the multispecific construct comprises: (i) H1 comprising the amino acid sequence of SEQ ID NO: 139, L1 comprising the amino acid sequence of SEQ ID NO: 122 or 207, H2 comprising the amino acid sequence of SEQ ID NO: 136, and L2 comprising the amino acid sequence of SEQ ID NO: 103; (ii) H1 comprising the amino acid sequence of SEQ ID NO: 138, L1 comprising the amino acid sequence of SEQ ID NO: 122 or 207, H2 comprising the amino acid sequence of SEQ ID NO: 137, and L2 comprising the amino acid sequence of SEQ ID NO: 103; (iii) H1 comprising the amino acid sequence of SEQ ID NO: 141, L1 comprising the amino acid sequence of SEQ ID NO: 122 or 207, H2 comprising the amino acid sequence of SEQ ID NO: 136, and L2 comprising the amino acid sequence of SEQ ID NO: NO: 103; (vi) H1 containing the amino acid sequence of SEQ ID NO: 140, L1 containing the amino acid sequence of SEQ ID NO: 122 or 207, H2 containing the amino acid sequence of SEQ ID NO: 137, and L2 containing the amino acid sequence of SEQ ID NO: 103; (v) H1 containing the amino acid sequence of SEQ ID NO: 126, L1 containing the amino acid sequence of SEQ ID NO: 120 or 208, H2 containing the amino acid sequence of SEQ ID NO: 136, and L2 containing the amino acid sequence of SEQ ID NO: 103; (vi) H1 containing the amino acid sequence of SEQ ID NO: 127, L1 containing the amino acid sequence of SEQ ID NO: 120 or 208, H2 containing the amino acid sequence of SEQ ID NO: 137, and L2 containing the amino acid sequence of SEQ ID NO: 103; (vii) H1 containing the amino acid sequence of SEQ ID NO: 127, L1 containing the amino acid sequence of SEQ ID NO: 120 or 208, H2 containing the amino acid sequence of SEQ ID NO: 137, and L2 containing the amino acid sequence of SEQ ID NO: 103 (viii) H1 containing the amino acid sequence of SEQ ID NO: 129, L1 containing the amino acid sequence of SEQ ID NO: 120 or 208, H2 containing the amino acid sequence of SEQ ID NO: 137, and L2 containing the amino acid sequence of SEQ ID NO: 103.

在一些實施例中，本文所述之治療發炎性疾病(例如，異位性皮膚炎、COPD或氣喘)的方法可達到一或多個以下生物活性：(1)抑制(例如，至少約10%、20%、30%、40%、50%、60%、70%、80%、90%或100%之任一者)免疫細胞成熟及/或活化(例如，B細胞、T輔助2型細胞、ILC2或APC (例如，樹突細胞))；(2)抑制(例如，至少約10%、20%、30%、40%、50%、60%、70%、80%、90%或100%之任一者)免疫細胞增生；(3)降低(例如，至少約10%、20%、30%、40%、50%、60%、70%、80%、90%或100%之任一者)全身或局部發炎水平；(4)緩解(例如，至少約10%、20%、30%、40%、50%、60%、70%、80%、90%或100%之任一者)發炎性疾病個體的一或多種症狀；(5)降低(例如，至少約10%、20%、30%、40%、50%、60%、70%、80%、90%或100%之任一者)全身或局部促發炎細胞激素水平(例如，TSLP、IL-13、CCL7、IL-6、IFN-γ、TNF-α)；(6)降低(例如，至少約10%、20%、30%、40%、50%、60%、70%、80%、90%或100%之任一者)發炎性疾病之表現、發生率或負荷；(7)延長發炎性免疫爆發的時間，諸如將個體發炎性疾病症狀消退後直至症狀重新出現的時間延長至少約1、2、4、6、12、18、20或更多小時，1、2、3、4、5，6、7或更多天，1、2、3、4或更多週，1、2、3、4、5、6、7、8、9、10、11、12、18或24個月或更長時間之任一者；(8)防止發炎性疾病的症狀再次出現持續至少約1、2、4、6、12、18、20或更多小時，1、2、3、4、5，6、7或更多天，1週、2週、3週、4週、5週、6週、7週、8週、9週、10週、11週、12週、1個月、2個月、3個月、4個月、5個月、6個月、7個月、8個月、9個月、10個月、11個月、12個月或更長時間之任一者；及(9)預防、抑制或降低(例如，至少約10%、20%、30%、40%、50%、60%、70%、80%、90%或100%之任一者)發炎性疾病復發的可能性。In some embodiments, the methods described herein for treating an inflammatory disease (e.g., atopic dermatitis, COPD, or asthma) can achieve one or more of the following biological activities: (1) inhibiting (e.g., at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) immune cell maturation and/or activation (e.g., B cells, T helper type 2 cells, ILC2, or APCs); (e.g., dendritic cells); (2) inhibit (e.g., by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) immune cell proliferation; (3) reduce (e.g., by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) systemic or local inflammation; (4) alleviate (e.g., by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) (5) reduction (e.g., by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) of systemic or local pro-inflammatory cytokine levels (e.g., TSLP, IL-13, CCL7, IL-6, IFN-γ, TNF-α); (6) reduction (e.g., by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or 100%) of the manifestations of an inflammatory disease , incidence, or burden; (7) prolonging the duration of an inflammatory immune outbreak, such as extending the time from the resolution of symptoms of an inflammatory disease in a subject until the reappearance of symptoms by at least about 1, 2, 4, 6, 12, 18, 20 or more hours, 1, 2, 3, 4, 5, 6, 7 or more days, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 18, or 24 months or longer; (8) preventing the reappearance of symptoms of an inflammatory disease for at least about 1, 2, 4, 6, 12, 18, 20 or more hours , 1, 2, 3, 4, 5, 6, 7 or more days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months or longer; and (9) preventing, inhibiting or reducing (e.g., by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90% or 100%) the likelihood of a recurrence of an inflammatory disease.

本文所提供之方法可在主要治療環境中實踐，亦即其中所進行之方法為主要/主程性治療(primary/definitive therapy)。在一些實施例中，該方法可在主要/主程性治療之前或與其結合進行。在一些實施例中，該方法用於治療先前接受過治療的個體(諸如人類)。本文所提供之任何治療方法皆可用於治療先前未接受過治療的個體(諸如人類)。在一些實施例中，該方法係用作一線療法。在一些實施例中，該方法係用作二線療法。The methods provided herein can be practiced in a primary treatment setting, i.e., where the method is performed as a primary/definitive therapy. In some embodiments, the methods can be performed prior to or in conjunction with a primary/definitive therapy. In some embodiments, the methods are used to treat a subject (e.g., a human) who has previously received treatment. Any of the treatment methods provided herein can be used to treat a subject (e.g., a human) who has not previously received treatment. In some embodiments, the methods are used as a first-line therapy. In some embodiments, the methods are used as a second-line therapy.

本文所述之方法適合治療多種發炎性疾病。該方法適用於所有階段的發炎性疾病，包括症狀出現後不久或處於緩解期的發炎性疾病。本文所述之方法可用作第一療法、第二療法、第三療法或與本領域已知之其他類型療法的組合療法，諸如抗發炎劑(例如，皮質類固醇)、基因療法、免疫療法、骨髓移植、幹細胞移植、標靶療法、營養療法等。在一些實施例中，該發炎性疾病對先前的療法沒有反應。The methods described herein are suitable for treating a variety of inflammatory diseases. The methods are applicable to inflammatory diseases at all stages, including those occurring shortly after symptoms appear or in remission. The methods described herein can be used as a primary therapy, a secondary therapy, a tertiary therapy, or in combination with other types of therapy known in the art, such as anti-inflammatory agents (e.g., corticosteroids), gene therapy, immunotherapy, bone marrow transplantation, stem cell transplantation, targeted therapy, nutritional therapy, and the like. In some embodiments, the inflammatory disease has not responded to previous treatments.

本文所述之多特異性構築體或分離的抗IL-13抗體構築體(或其醫藥組成物)之任一者的示例性投予途徑包括但不限於：靜脈內、腔內、動脈內、肌內、皮下、非經口、透皮或腹膜內途徑，或遞輸至受到發炎性疾病影響的淋巴腺、體腔、器官或組織。在一些實施例中，藉由靜脈內(諸如輸注或推注)來投予多特異性構築體或分離的抗IL-13抗體構築體或其醫藥組成物。在一些實施例中，藉由皮下投予多特異性構築體或分離的抗IL-13抗體構築體或其醫藥組成物。Exemplary routes of administration of any of the multispecific constructs or isolated anti-IL-13 antibody constructs (or pharmaceutical compositions thereof) described herein include, but are not limited to, intravenous, intracavitary, intraarterial, intramuscular, subcutaneous, parenteral, transdermal, or intraperitoneal routes, or delivery to a lymph node, body cavity, organ, or tissue affected by an inflammatory disease. In some embodiments, the multispecific construct or isolated anti-IL-13 antibody construct or pharmaceutical composition thereof is administered intravenously (e.g., by infusion or bolus injection). In some embodiments, the multispecific construct or isolated anti-IL-13 antibody construct or pharmaceutical composition thereof is administered subcutaneously.

在一些實施例中，藉由靜脈內輸注，以任何合適速率投予本文所述之多特異性構築體或分離的抗IL-13抗體構築體(或其醫藥組成物)。In some embodiments, a multispecific construct or isolated anti-IL-13 antibody construct described herein (or pharmaceutical composition thereof) is administered by intravenous infusion at any suitable rate.

向個體(諸如人類)投予的本文所述之多特異性構築體或分離的抗IL-13抗體構築體或其醫藥組成物的給藥方案可能因特定組成物、投予方法及待治療之發炎性疾病的特定類型而變。在一些實施例中，該多特異性構築體或分離的抗IL-13抗體構築體或其醫藥組成物之有效量低於引發毒性作用的量(亦即，高於臨床上可接受之毒性水平的作用)或當向個體投予組成物時其潛在副作用可處於可控或耐受的水平。The dosing regimen of a multispecific construct or isolated anti-IL-13 antibody construct described herein, or a pharmaceutical composition thereof, administered to a subject (e.g., a human) may vary depending on the specific composition, the method of administration, and the specific type of inflammatory disease to be treated. In some embodiments, the effective amount of the multispecific construct or isolated anti-IL-13 antibody construct or a pharmaceutical composition thereof is below the amount that causes toxic effects (i.e., effects above a clinically acceptable level of toxicity) or the potential side effects are manageable or tolerable when the composition is administered to a subject.

本文所述之多特異性構築體或分離的抗IL-13抗體構築體(或其醫藥組成物)的有效量可以單劑量或多劑量方式投予(例如，靜脈內或皮下)。針對包含以多劑量投予多特異性構築體或分離的抗IL-13抗體構築體(或其醫藥組成物)的方法，示例性給藥頻率包括但不限於：每小時、每天、每天不間斷、每週、每週不間斷、三週中的兩週每週一次、四週中的三週每週一次、每三週一次、每兩週一次、每月一次、每六個月、每年等。在一些實施例中，該多特異性構築體或分離的抗IL-13抗體構築體(或其醫藥組成物)的投予為約每2週一次、每3週一次，每4週一次，每6週一次或每8週一次。在一些實施例中，該多特異性構築體或分離的抗IL-13抗體構築體(或其醫藥組成物)的投予為每週至少約1次、2次、3次、4次、5次、6次或7次(亦即，每天)之任一者。在一些實施例中，每次投予之間的間隔為小於約3年、2年、1年、12個月、11個月、10個月、9個月、8個月、7個月、6個月、5個月、4個月、3個月、2個月、1個月、5週、4週、3週、2週、1週、7天、6天、5天、4天、3天、2天或1天之任一者。在一些實施例中，每次投予之間的間隔為大於約每次投予之間的間隔為小於1週、2週、3週、4週、5週、1個月、2個月、3個月、4個月、5個月、6個月、7個月、8個月、9個月、10個月、11個月、12個月、1年、2年或3年之任一者。在一些實施例中，給藥時間表沒有間斷。An effective amount of a multispecific construct or isolated anti-IL-13 antibody construct (or pharmaceutical composition thereof) described herein can be administered in a single dose or in multiple doses (e.g., intravenously or subcutaneously). For methods comprising administering a multispecific construct or isolated anti-IL-13 antibody construct (or pharmaceutical composition thereof) in multiple doses, exemplary dosing frequencies include, but are not limited to, hourly, daily, continuously daily, weekly, continuously weekly, once a week for two out of three weeks, once a week for three out of four weeks, once every three weeks, once every two weeks, once a month, every six months, annually, and the like. In some embodiments, the multispecific construct or isolated anti-IL-13 antibody construct (or pharmaceutical composition thereof) is administered about once every 2 weeks, once every 3 weeks, once every 4 weeks, once every 6 weeks, or once every 8 weeks. In some embodiments, the multispecific construct or isolated anti-IL-13 antibody construct (or pharmaceutical composition thereof) is administered at least about once, twice, three times, four times, five times, six times, or seven times per week (i.e., every day). In some embodiments, the intervals between each administration are less than about any of 3 years, 2 years, 1 year, 12 months, 11 months, 10 months, 9 months, 8 months, 7 months, 6 months, 5 months, 4 months, 3 months, 2 months, 1 month, 5 weeks, 4 weeks, 3 weeks, 2 weeks, 1 week, 7 days, 6 days, 5 days, 4 days, 3 days, 2 days, or 1 day. In some embodiments, the intervals between each administration are greater than about any of 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 2 years, or 3 years. In some embodiments, there are no breaks in the dosing schedule.

本文所述之多特異性構築體或分離的抗IL-13抗體構築體(或其醫藥組成物)的投予可延長一段較長的時間，諸如1天至約一週、約一週至約一個月、約一個月至約一年、約一年至約幾年。在一些實施例中，該多特異性構築體或分離的抗IL-13抗體構築體(或其醫藥組成物)係投予(例如，靜脈內或皮下)持續至少約1天、2天、3天、4天、5天、6天、7天、1週、2週、3週、4週、5週、1個月、2個月、3個月、4個月、5個月、6個月、7個月、8個月、9個月、10個月、11個月、12個月、1年、2年、3年、4年或更多之任一者的時間。 示例性實施例Administration of the multispecific constructs or isolated anti-IL-13 antibody constructs described herein (or pharmaceutical compositions thereof) can be extended over a prolonged period of time, such as from 1 day to about 1 week, from about 1 week to about 1 month, from about 1 month to about 1 year, or from about 1 year to about several years. In some embodiments, the multispecific construct or isolated anti-IL-13 antibody construct (or pharmaceutical composition thereof) is administered (e.g., intravenously or subcutaneously) for at least about 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, 1 year, 2 years, 3 years, 4 years, or more.Exemplary Embodiments

實施例1. 一種多特異性構築體，其包含： (1)特異性地結合至介白素-13 (IL-13)的第一抗體部分，以及 (2)特異性地結合至第二抗原的第二抗體部分。Example 1. A multispecific construct comprising:(1) a first antibody portion that specifically binds to interleukin-13 (IL-13), and(2) a second antibody portion that specifically binds to a second antigen.

實施例2. 如實施例1之多特異性構築體，其中該第二抗原為由免疫細胞產生的蛋白質。Example 2. The multispecific construct of Example 1, wherein the second antigen is a protein produced by an immune cell.

實施例3. 如實施例1或2之多特異性構築體，其中該第二抗原為胸腺基質淋巴球生成素(TSLP)。Embodiment 3. The multispecific construct of embodiment 1 or 2, wherein the second antigen is thymic stromal lymphopoietin (TSLP).

實施例4. 如實施例1至3中任一者之多特異性構築體，其中該第一抗體部分包含重鏈可變區(VH1)及輕鏈可變區(VL1)，其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多3個胺基酸變異的變體，(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多3個胺基酸變異的變體，以及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多3個胺基酸變異的變體，且該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1或其含有至多3個胺基酸變異的變體，(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多3個胺基酸變異的變體，以及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多3個胺基酸變異的變體。Embodiment 4. The multispecific construct of any one of Embodiments 1 to 3, wherein the first antibody portion comprises a heavy chain variable region (VH1) and a light chain variable region (VL1), wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66 or a variant thereof containing up to 3 amino acid variations, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67 or a variant thereof containing up to 3 amino acid variations, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68 or a variant thereof containing up to 3 amino acid variations, and the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70 or a variant thereof containing up to 3 amino acid variations, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67 or a variant thereof containing up to 3 amino acid variations, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68 or a variant thereof containing up to 3 amino acid variations. (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to 3 amino acid variations, and (iv) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to 3 amino acid variations.

實施例5. 如實施例4之多特異性構築體，其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1，(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2，以及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3，且該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1，(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3。Example 5. The multispecific construct of Example 4, wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, and the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70, (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72.

實施例6. 如實施例4或5之多特異性構築體，其中該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80%序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80%序列同一性的變體。Embodiment 6. The multispecific construct of Embodiment 4 or 5, wherein the VH1 comprises the amino acid sequence of SEQ ID NO: 65 or a variant thereof having at least about 80% sequence identity with SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69 or a variant thereof having at least about 80% sequence identity with SEQ ID NO: 69.

實施例7. 如實施例4至6中任一者之多特異性構築體，其中該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。Embodiment 7. The multispecific construct of any one of Embodiments 4 to 6, wherein the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69.

實施例8. 如實施例1至7中任一者之多特異性構築體，其中該第一抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、sdAb及scFv。Embodiment 8. The multispecific construct of any one of embodiments 1 to 7, wherein the first antibody portion is selected from the group consisting of: full-length antibody, Fab, Fab', F(ab')2, sdAb and scFv.

實施例9. 如實施例8之多特異性構築體，其中該第一抗體部分為scFv (「抗IL-13 scFv」)。Embodiment 9. The multispecific construct of embodiment 8, wherein the first antibody portion is a scFv ("anti-IL-13 scFv").

實施例10.    如實施例9之多特異性構築體，其中該抗IL-13 scFv含有SEQ ID NO:108之胺基酸序列。Example 10. The multispecific construct of Example 9, wherein the anti-IL-13 scFv comprises the amino acid sequence of SEQ ID NO: 108.

實施例11.    如實施例8之多特異性構築體，其中該第一抗體部分為Fab (「抗IL-13 Fab」)。Example 11. The multispecific construct of Example 8, wherein the first antibody portion is Fab ("anti-IL-13 Fab").

實施例12.    如實施例11之多特異性構築體，其中該抗IL-13 Fab包含含有SEQ ID NO:124之胺基酸序列的第一多肽及含有SEQ ID NO:102或197之胺基酸序列的第二多肽。Example 12. The multispecific construct of Example 11, wherein the anti-IL-13 Fab comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197.

實施例13.    如實施例8之多特異性構築體，其中該第一抗體部分為全長抗體(「抗IL-13全長抗體」)。Example 13. A multispecific construct as in Example 8, wherein the first antibody portion is a full-length antibody ("anti-IL-13 full-length antibody").

實施例14.    如實施例13之多特異性構築體，其中該抗IL-13全長抗體包含衍生自人類IgG1、IgG2或IgG4的Fc域。Example 14. The multispecific construct of Example 13, wherein the anti-IL-13 full-length antibody comprises an Fc domain derived from human IgG1, IgG2 or IgG4.

實施例15.    如實施例14之多特異性構築體，其中該Fc域係衍生自人類IgG1，且其中： i)該Fc域之第一次單元及第二次單元各自含有SEQ ID NO:76-79之任一者的胺基酸序列； ii)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列； iii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列； iv)該Fc域之第一次單元含有SEQ ID NO:95之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:96之胺基酸序列；或 v)該Fc域之第一次單元含有SEQ ID NO:96之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:95之胺基酸序列。Example 15. The multispecific construct of Example 14, wherein the Fc domain is derived from human IgG1, and wherein:i) the first and second subunits of the Fc domain each contain the amino acid sequence of any one of SEQ ID NOs: 76-79;ii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81;iii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80;iv) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 95, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 96; orv) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: The amino acid sequence of SEQ ID NO:96 is present, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO:95.

實施例16.    如實施例13至15中任一者之多特異性構築體，其中該抗IL-13全長抗體包含： i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； ii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； iii)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈； iv)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； v)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； vi)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈； vii)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； viii)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； ix)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或 x)含有SEQ ID NO:130之胺基酸序列的第一重鏈、含有SEQ ID NO:131之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。Example 16. The multispecific construct of any one of Examples 13 to 15, wherein the full-length anti-IL-13 antibody comprises:i) two heavy chains each containing the amino acid sequence of SEQ ID NO: 125 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197;ii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 101 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197;iii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 123 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206;iv) two heavy chains each containing the amino acid sequence of SEQ ID NO: 209 and two light chains each containing the amino acid sequence of SEQ ID NO: NO:122 or 207;v) two heavy chains each containing the amino acid sequence of SEQ ID NO:210 and two light chains each containing the amino acid sequence of SEQ ID NO:122 or 207;vi) two heavy chains each containing the amino acid sequence of SEQ ID NO:211 and two light chains each containing the amino acid sequence of SEQ ID NO:121 or 206;vii) two heavy chains each containing the amino acid sequence of SEQ ID NO:212 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208;viii) two heavy chains each containing the amino acid sequence of SEQ ID NO:213 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208;ix) two heavy chains each containing the amino acid sequence of SEQ ID NO:214 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208; A heavy chain containing the amino acid sequence of SEQ ID NO: 225 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197; orx) a first heavy chain containing the amino acid sequence of SEQ ID NO: 130, a second heavy chain containing the amino acid sequence of SEQ ID NO: 131, and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197.

實施例17.    如實施例3至16中任一者之多特異性構築體，其中該第二抗體部分包含重鏈可變區(VH2)及輕鏈可變區(VL2)，其中： (a)該VH2包含(i)含有SEQ ID NO:2之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:3之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3； (b)該VH2包含(i)含有SEQ ID NO:12之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:13之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3； (c)該VH2包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3； (d)該VH2包含(i)含有SEQ ID NO:28之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:29之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:30之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:32之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:33之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:34之胺基酸序列的CDR-L3； (e)該VH2包含(i)含有SEQ ID NO:36之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:37之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:38之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:40之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:41之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:42之胺基酸序列的CDR-L3；或 (f)該VH2包含(i)含有SEQ ID NO:44之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:45之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:46之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:48之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:49之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:50之胺基酸序列的CDR-L3。Example 17. The multispecific construct of any one of Examples 3 to 16, wherein the second antibody portion comprises a heavy chain variable region (VH2) and a light chain variable region (VL2), wherein:(a) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 2; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 3; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 4; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 6; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 7; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 8;(b) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 12; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 13; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 14. NO:13; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:4; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:6; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:7; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:8;(c) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:22; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:25; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: NO:26;(d) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:28; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:29; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:30; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:32; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:33; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:34;(e) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:36; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:37; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:38. (i) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:38; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:40; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:41; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:42; or(f) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:44; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:45; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:46; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:48; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:49; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:50.

實施例18.    如實施例之多特異性構築體17，其中： (a)該VH2含有SEQ ID NO:1之胺基酸序列，或與SEQ ID NO:1具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:5之胺基酸序列，或與SEQ ID NO:5具有至少約80%序列同一性的胺基酸序列； (b)該VH2含有SEQ ID NO:9之胺基酸序列，或與SEQ ID NO:9具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:10之胺基酸序列，或與SEQ ID NO:10具有至少約80%序列同一性的胺基酸序列； (c)該VH2含有SEQ ID NO:11之胺基酸序列，或與SEQ ID NO:11具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:15之胺基酸序列，或與SEQ ID NO:15具有至少約80%序列同一性的胺基酸序列； (d)該VH2含有SEQ ID NO:19之胺基酸序列，或與SEQ ID NO:19具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:23之胺基酸序列，或與SEQ ID NO:23具有至少約80%序列同一性的胺基酸序列； (e)該VH2含有SEQ ID NO:27之胺基酸序列，或與SEQ ID NO:27具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:31之胺基酸序列，或與SEQ ID NO:31具有至少約80%序列同一性的胺基酸序列； (f)該VH2含有SEQ ID NO:35之胺基酸序列，或與SEQ ID NO:35具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:39之胺基酸序列，或與SEQ ID NO:39具有至少約80%序列同一性的胺基酸序列； (g)該VH2含有SEQ ID NO:43之胺基酸序列，或與SEQ ID NO:43具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:47之胺基酸序列，或與SEQ ID NO:47具有至少約80%序列同一性的胺基酸序列； (h)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (i)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (j)該VH2含有SEQ ID NO:56之胺基酸序列，或與SEQ ID NO:56具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (k)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列； (l)該VH2含有SEQ ID NO:56之胺基酸序列，或與SEQ ID NO:56具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列； (m)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列； (n)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列； (o)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (p)該VH2含有SEQ ID NO:58之胺基酸序列，或與SEQ ID NO:58具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (q)該VH2含有SEQ ID NO:60之胺基酸序列，或與SEQ ID NO:60具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (r)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (s)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (t)該VH2含有SEQ ID NO:57之胺基酸序列，或與SEQ ID NO:57具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (u)該VH2含有SEQ ID NO:58之胺基酸序列，或與SEQ ID NO:58具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (v)該VH2含有SEQ ID NO:62之胺基酸序列，或與SEQ ID NO:62具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (w)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (x)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (y)該VH2含有SEQ ID NO:61之胺基酸序列，或與SEQ ID NO:61具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (z)該VH2含有SEQ ID NO:62之胺基酸序列，或與SEQ ID NO:62具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (aa)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (bb)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (cc)該VH2含有SEQ ID NO:188之胺基酸序列，或與SEQ ID NO:188具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:189之胺基酸序列，或與SEQ ID NO:189具有至少約80%序列同一性的胺基酸序列； (dd)該VH2含有SEQ ID NO:188之胺基酸序列，或與SEQ ID NO:188具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:190之胺基酸序列，或與SEQ ID NO:190具有至少約80%序列同一性的胺基酸序列；或 (ee)該VH2含有SEQ ID NO:228之胺基酸序列，或與SEQ ID NO:228具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:190之胺基酸序列，或與SEQ ID NO:190具有至少約80%序列同一性的胺基酸序列。Example 18. The multispecific construct 17 of Example 1, wherein:(a) the VH2 comprises the amino acid sequence of SEQ ID NO:1, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:1, and the VL2 comprises the amino acid sequence of SEQ ID NO:5, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:5;(b) the VH2 comprises the amino acid sequence of SEQ ID NO:9, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:9, and the VL2 comprises the amino acid sequence of SEQ ID NO:10, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:10;(c) the VH2 comprises the amino acid sequence of SEQ ID NO:11, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:11, and the VL2 comprises the amino acid sequence of SEQ ID NO:11. NO:15, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:15;(d) the VH2 comprises the amino acid sequence of SEQ ID NO:19, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:19, and the VL2 comprises the amino acid sequence of SEQ ID NO:23, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:23;(e) the VH2 comprises the amino acid sequence of SEQ ID NO:27, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:27, and the VL2 comprises the amino acid sequence of SEQ ID NO:31, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:31;(f) the VH2 comprises the amino acid sequence of SEQ ID NO:35, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:36. (g) the VH2 comprises the amino acid sequence of SEQ ID NO:43, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:43, and the VL2 comprises the amino acid sequence of SEQ ID NO:47, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:47; (h) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:53; (i) the VH2 comprises the amino acid sequence of SEQ ID NO: (i) the VH2 comprises the amino acid sequence of SEQ ID NO:56, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:56, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:52; (ii) the VH2 comprises the amino acid sequence of SEQ ID NO:55, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:55, and the VL2 comprises the amino acid sequence of SEQ ID NO:51, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:51; (l) the VH2 comprises the amino acid sequence of SEQ ID NO:56, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:56, and the VL2 comprises the amino acid sequence of SEQ ID NO:51, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:51;(m) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:51, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:51;(n) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:51, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: NO:51 has an amino acid sequence with at least about 80% sequence identity;(o) the VH2 contains the amino acid sequence of SEQ ID NO:55, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:55, and the VL2 contains the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:52;(p) the VH2 contains the amino acid sequence of SEQ ID NO:58, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:58, and the VL2 contains the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:52;(q) the VH2 contains the amino acid sequence of SEQ ID NO:60, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:60, and the VL2 contains the amino acid sequence of SEQ ID NO: NO:52, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:52;(r) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:52;(s) the VH2 comprises the amino acid sequence of SEQ ID NO:55, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:55, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:53;(t) the VH2 comprises the amino acid sequence of SEQ ID NO:57, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:58. NO:57 has an amino acid sequence with at least about 80% sequence identity, and the VL2 contains the amino acid sequence of SEQ ID NO:53, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:53;(u) the VH2 contains the amino acid sequence of SEQ ID NO:58, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:58, and the VL2 contains the amino acid sequence of SEQ ID NO:53, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:53;(v) the VH2 contains the amino acid sequence of SEQ ID NO:62, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:62, and the VL2 contains the amino acid sequence of SEQ ID NO:53, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:53;(w) the VH2 contains the amino acid sequence of SEQ ID NO: (x) the VH2 comprises the amino acid sequence of SEQ ID NO:55, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:55, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:54; (y) the VH2 comprises the amino acid sequence of SEQ ID NO:61, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:61, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:54; (z) the VH2 comprises the amino acid sequence of SEQ ID NO:62, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:62, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:54;(aa) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:54;(bb) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: NO:52 has an amino acid sequence with at least about 80% sequence identity;(cc) the VH2 contains the amino acid sequence of SEQ ID NO:188, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:188, and the VL2 contains the amino acid sequence of SEQ ID NO:189, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:189;(dd) the VH2 contains the amino acid sequence of SEQ ID NO:188, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:188, and the VL2 contains the amino acid sequence of SEQ ID NO:190, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:190; or(ee) the VH2 contains the amino acid sequence of SEQ ID NO:228, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:228, and the VL2 contains the amino acid sequence of SEQ ID NO: An amino acid sequence of SEQ ID NO: 190, or an amino acid sequence having at least about 80% sequence identity with SEQ ID NO: 190.

實施例19.    如實施例1至18中任一者之多特異性構築體，其中該第二抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、sdAb及scFv。Embodiment 19. The multispecific construct of any one of embodiments 1 to 18, wherein the second antibody portion is selected from the group consisting of: a full-length antibody, Fab, Fab', F(ab')2 , sdAb, and scFv.

實施例20.    如實施例19之多特異性構築體，其中該第二抗體部分為全長抗體。Example 20. The multispecific construct of Example 19, wherein the second antibody portion is a full-length antibody.

實施例21.    如實施例20之多特異性構築體，其中該全長抗體包含衍生自人類IgG1、IgG2或IgG4的Fc域。Example 21. The multispecific construct of Example 20, wherein the full-length antibody comprises an Fc domain derived from human IgG1, IgG2 or IgG4.

實施例22.    如實施例21之多特異性構築體，其中該Fc域係衍生自人類IgG1，且其中： i)該Fc域之第一次單元及第二次單元各自含有SEQ ID NO:76-79之任一者的胺基酸序列； ii)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列；或 iii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列。Example 22. The multispecific construct of Example 21, wherein the Fc domain is derived from human IgG1, and wherein:i) the first and second subunits of the Fc domain each contain the amino acid sequence of any one of SEQ ID NOs: 76-79;ii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81; oriii) the first subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 81, and the second subunit of the Fc domain contains the amino acid sequence of SEQ ID NO: 80.

實施例23.    如實施例20至22中任一者之多特異性構築體，其中該全長抗體為抗TSLP全長抗體，其包含： i)兩條各自含有SEQ ID NO:104之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈； ii)兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈；或 iii)含有SEQ ID NO:136之胺基酸序列的第一重鏈、含有SEQ ID NO:137之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈。Example 23. The multispecific construct of any one of Examples 20 to 22, wherein the full-length antibody is an anti-TSLP full-length antibody comprising:i) two heavy chains each containing the amino acid sequence of SEQ ID NO: 104 and two light chains each containing the amino acid sequence of SEQ ID NO: 103;ii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 105 and two light chains each containing the amino acid sequence of SEQ ID NO: 103; oriii) a first heavy chain each containing the amino acid sequence of SEQ ID NO: 136, a second heavy chain each containing the amino acid sequence of SEQ ID NO: 137, and two light chains each containing the amino acid sequence of SEQ ID NO: 103.

實施例24.    如實施例19之多特異性構築體，其中該第二抗體部分為scFv。Example 24. The multispecific construct of Example 19, wherein the second antibody portion is a scFv.

實施例25.    如實施例24之多特異性構築體，其中該第二抗體部分為含有SEQ ID NO:106、107、218、219、226、227及229之任一者之胺基酸序列的抗TSLP scFv。Example 25. The multispecific construct of Example 24, wherein the second antibody portion is an anti-TSLP scFv comprising the amino acid sequence of any one of SEQ ID NOs: 106, 107, 218, 219, 226, 227, and 229.

實施例26.    如實施例19之多特異性構築體，其中該第二抗體部分為Fab。Example 26. The multispecific construct of Example 19, wherein the second antibody portion is Fab.

實施例27.    如實施例26之多特異性構築體，其中該第二抗體部分為抗TSLP Fab，其中該抗TSLP Fab包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。Example 27. The multispecific construct of Example 26, wherein the second antibody portion is an anti-TSLP Fab, wherein the anti-TSLP Fab comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103.

實施例28.    如實施例1至7及17至27中任一者之多特異性構築體，其中該第一抗體部分包含重鏈(H1)及輕鏈(L1)，其中該H1包含VH1及H1-CH1，其中該L1包含VL1及L1-CL，且其中： (i)該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)；及/或 (ii)該H1包含位於位置126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。Embodiment 28. The multispecific construct of any one of Embodiments 1 to 7 and 17 to 27, wherein the first antibody portion comprises a heavy chain (H1) and a light chain (L1), wherein the H1 comprises VH1 and H1-CH1, wherein the L1 comprises VL1 and L1-CL, and wherein:(i) the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering); and/or(ii) the H1 comprises substitutions at positions 126 and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering).

實施例29.    如實施例28之多特異性構築體，其中該H1包含位於位置F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。Example 29. The multispecific construct of Example 28, wherein the H1 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering).

實施例30.    如實施例29之多特異性構築體，其中該F170取代為F170I或F170V、該S183取代為S183L或S183I且該V185取代為V185L。Example 30. The multispecific construct of Example 29, wherein the F170 is replaced by F170I or F170V, the S183 is replaced by S183L or S183I, and the V185 is replaced by V185L.

實施例31.    如實施例29或30之多特異性構築體，其中該L135取代為L135F。Example 31. The multispecific construct of Example 29 or 30, wherein L135 is replaced by L135F.

實施例32.    如實施例29至31中任一者之多特異性構築體，其中： (a)  該H1包含F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)；或 (b)  該H1包含F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。Embodiment 32. The multispecific construct of any one of Embodiments 29 to 31, wherein:(a) H1 comprises F170I, S183L, and V185L substitutions (EU numbering), and L1 comprises L135F substitution (EU numbering); or(b) H1 comprises F170V, S183I, and V185L substitutions (EU numbering), and L1 comprises L135F substitution (EU numbering).

實施例33.    如實施例28至32中任一者之多特異性構築體，其中該L1係衍生自λ輕鏈。Embodiment 33. The multispecific construct of any one of embodiments 28 to 32, wherein L1 is derived from a lambda light chain.

實施例34.    如實施例33之多特異性構築體，其中該λ輕鏈含有SEQ ID NO:74或75之胺基酸序列。Example 34. The multispecific construct of Example 33, wherein the λ light chain contains the amino acid sequence of SEQ ID NO: 74 or 75.

實施例35.    如實施例33之多特異性構築體，其中該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。Example 35. The multispecific construct of Example 33, wherein the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering).

實施例36.    如實施例35之多特異性構築體，其中該H1包含F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。Example 36. The multispecific construct of Example 35, wherein the H1 comprises F126C and C220S substitutions (EU numbering), and the L1 comprises E124C and C214S substitutions (EU numbering).

實施例37.    如實施例28至32中任一者之多特異性構築體，其中該L1係衍生自κ輕鏈。Embodiment 37. The multispecific construct of any one of embodiments 28 to 32, wherein L1 is derived from a kappa light chain.

實施例38.    如實施例37之多特異性構築體，其中該κ輕鏈含有SEQ ID NO:73之胺基酸序列。Example 38. The multispecific construct of Example 37, wherein the κ light chain contains the amino acid sequence of SEQ ID NO:73.

實施例39.    如實施例37之多特異性構築體，其中該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。Example 39. The multispecific construct of Example 37, wherein the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering).

實施例40.    如實施例39之多特異性構築體，其中該H1包含F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。Example 40. The multispecific construct of Example 39, wherein the H1 comprises F126C and C220S substitutions (EU numbering), and the L1 comprises Q124C and C214S substitutions (EU numbering).

實施例41.    如實施例28至33、35至37、39及40中任一者之多特異性構築體，其中： (i)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代； (ii)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代； (iii)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含Q124C、L135F及C214S取代；或 (iv)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含Q124C、L135F及C214S取代；且 其中該位置係根據EU編號。Embodiment 41. The multispecific construct of any one of Embodiments 28 to 33, 35 to 37, 39, and 40, wherein:(i) H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and L1 comprises E124C, L135F, and C214S substitutions;(ii) H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and L1 comprises E124C, L135F, and C214S substitutions;(iii) H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and L1 comprises Q124C, L135F, and C214S substitutions; or (iv) the H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and the L1 comprises Q124C, L135F, and C214S substitutions; andwherein the positions are according to EU numbering.

實施例42.    如實施例1至41中任一者之多特異性構築體，其進一步包含Fc域，其中該Fc域包含第一次單元及第二次單元。Embodiment 42. The multispecific construct of any one of Embodiments 1 to 41, further comprising an Fc domain, wherein the Fc domain comprises a first unit and a second unit.

實施例43.    如實施例42之多特異性構築體，其中該Fc域係衍生自IgG，該IgG選自由以下組成之群組：IgG1、IgG2、IgG3及IgG4。Example 43. The multispecific construct of Example 42, wherein the Fc domain is derived from IgG, and the IgG is selected from the group consisting of: IgG1, IgG2, IgG3 and IgG4.

實施例44.    如實施例43之多特異性構築體，其中該Fc域係衍生自IgG1。Example 44. The multispecific construct of Example 43, wherein the Fc domain is derived from IgG1.

實施例45.    如實施例44之多特異性構築體，其中該Fc域之每一次單元包含L234A及L235A取代(EU編號)。Example 45. The multispecific construct of Example 44, wherein each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering).

實施例46.    如實施例45之多特異性構築體，其中該Fc域之每一次單元含有SEQ ID NO:77之胺基酸序列。Example 46. The multispecific construct of Example 45, wherein each subunit of the Fc domain contains the amino acid sequence of SEQ ID NO:77.

實施例47.    如實施例44或45之多特異性構築體，其中該Fc域包含M428L及N434S取代(EU編號)。Embodiment 47. The multispecific construct of embodiment 44 or 45, wherein the Fc domain comprises M428L and N434S substitutions (EU numbering).

實施例48.    如實施例47之多特異性構築體，其中該Fc域之每一次單元含有SEQ ID NO:78之胺基酸序列。Example 48. The multispecific construct of Example 47, wherein each subunit of the Fc domain contains the amino acid sequence of SEQ ID NO:78.

實施例49.    如實施例44、45及47中任一者之多特異性構築體，其中該Fc域包含M252Y、S254T及T256E取代(EU編號)。Embodiment 49. The multispecific construct of any one of Embodiments 44, 45 and 47, wherein the Fc domain comprises M252Y, S254T and T256E substitutions (EU numbering).

實施例50.    如實施例49之多特異性構築體，其中該Fc域之每一次單元含有SEQ ID NO:79之胺基酸序列。Example 50. The multispecific construct of Example 49, wherein each subunit of the Fc domain contains the amino acid sequence of SEQ ID NO:79.

實施例51.     如實施例44、45、47及49中任一者之多特異性構築體，其中： (i)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或 (ii)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。Embodiment 51. The multispecific construct of any one of Embodiments 44, 45, 47, and 49, wherein:(i) the first Fc domain subunit comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second Fc domain subunit comprises T366W substitution (EU numbering); or(ii) the second Fc domain subunit comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first Fc domain subunit comprises T366W substitution (EU numbering).

實施例52.    如實施例51之多特異性構築體，其中： (i)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列； (ii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列； (iii)該Fc域之第一次單元含有SEQ ID NO:95之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:96之胺基酸序列；或 (iv)該Fc域之第一次單元含有SEQ ID NO:96之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:95之胺基酸序列。Example 52. The multispecific construct of Example 51, wherein:(i) the first subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:80, and the second subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:81;(ii) the first subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:81, and the second subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:80;(iii) the first subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:95, and the second subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:96; or(iv) the first subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:96, and the second subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:95.

實施例53.    如實施例1至7、17、18及28至52中任一者之多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含： i)含有VL1及L1-CL的第一輕鏈(L1)； ii)含有VH1、H1-CH1及Fc域之第一次單元的第一重鏈(H1)； iii)含有VH2、H2-CH1及該Fc域之第二次單元的第二重鏈(H2)；及 iv)含有VL2及L2-CL的第二輕鏈(L2)；且 其中H1及L1形成抗IL-13抗體部分，且H2及L2形成該第二抗體部分。Embodiment 53. The multispecific construct of any one of Embodiments 1 to 7, 17, 18, and 28 to 52, wherein the multispecific construct is a heterodimeric bispecific antibody comprising:i) a first light chain (L1) comprising VL1 and L1-CL;ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain;iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and the Fc domain; andiv) a second light chain (L2) comprising VL2 and L2-CL; andwherein H1 and L1 form the anti-IL-13 antibody portion, and H2 and L2 form the second antibody portion.

實施例54.    如實施例53之多特異性構築體，其中該Fc域係衍生自人類IgG1，其中該L1-CL係衍生自人類λ輕鏈，且其中： (a)該H1包含F170I、S183L及V185L取代，且該L1包含L135F取代； (b)該H1包含F170V、S183I及V185L取代，且該L1包含L135F取代； (c)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代； (d)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代；或 (e)該H1包含F126C及C220S取代，且該L1包含E124C及C214S取代；且 其中該胺基酸位置係根據EU編號。Example 54. The multispecific construct of Example 53, wherein the Fc domain is derived from human IgG1, wherein the L1-CL is derived from a human lambda light chain, and wherein:(a) the H1 comprises F170I, S183L, and V185L substitutions, and the L1 comprises an L135F substitution;(b) the H1 comprises F170V, S183I, and V185L substitutions, and the L1 comprises an L135F substitution;(c) the H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and the L1 comprises E124C, L135F, and C214S substitutions; (d) H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and L1 comprises E124C, L135F, and C214S substitutions; or(e) H1 comprises F126C and C220S substitutions, and L1 comprises E124C and C214S substitutions; andwherein the amino acid positions are according to EU numbering.

實施例55.    如實施例53或54之多特異性構築體，其中該Fc域係衍生自人類IgG1，其中該L1-CL係衍生自人類λ輕鏈，且其中： (a)該H1包含F170I、S183L、V185L、T366S、L368A及Y407V取代，該L1包含L135F取代，且該H2包含T366W取代； (b)該H1包含F170I、S183L、V185L及T366W取代，該L1包含L135F取代，且該H2包含T366S、L368A及Y407V取代； (c)該H1包含F170V、S183I、V185L、T366S、L368A及Y407V取代，該L1包含L135F取代，且該H2包含T366W取代； (d)該H1包含F170V、S183I、V185L及T366W取代，該L1包含L135F取代，且該H2包含T366S、L368A及Y407V取代； (e)該H1包含F126C、F170I、S183L、V185L、C220S、T366S、L368A及Y407V取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366W取代； (f)該H1包含F126C、F170I、S183L、V185L、C220S及T366W取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366S、L368A及Y407V取代； (g)該H1包含F126C、F170V、S183I、V185L、C220S、T366S、L368A及Y407V取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366W取代； (h)該H1包含F126C、F170V、S183I、V185L、C220S及T366W取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366S、L368A及Y407V取代； (i)該H1包含F126C、C220S、T366S、L368A及Y407V取代，該L1包含E124C及C214S取代，且該H2包含T366W取代；或 (j)該H1包含F126C、C220S及T366W取代，該L1包含E124C及C214S取代，且該H2包含T366S、L368A及Y407V取代；且 其中該胺基酸位置係根據EU編號。Example 55. The multispecific construct of Example 53 or 54, wherein the Fc domain is derived from human IgG1, wherein the L1-CL is derived from a human lambda light chain, and wherein:(a) H1 comprises F170I, S183L, V185L, T366S, L368A, and Y407V substitutions, L1 comprises L135F substitution, and H2 comprises T366W substitution;(b) H1 comprises F170I, S183L, V185L, and T366W substitutions, L1 comprises L135F substitution, and H2 comprises T366S, L368A, and Y407V substitutions; (c) H1 comprises F170V, S183I, V185L, T366S, L368A, and Y407V substitutions, L1 comprises L135F substitutions, and H2 comprises T366W substitutions;(d) H1 comprises F170V, S183I, V185L, and T366W substitutions, L1 comprises L135F substitutions, and H2 comprises T366S, L368A, and Y407V substitutions;(e) H1 comprises F126C, F170I, S183L, V185L, C220S, T366S, L368A, and Y407V substitutions, L1 comprises E124C, L135F, and C214S substitutions, and H2 comprises T366W substitutions; (f) H1 comprises F126C, F170I, S183L, V185L, C220S, and T366W substitutions, L1 comprises E124C, L135F, and C214S substitutions, and H2 comprises T366S, L368A, and Y407V substitutions;(g) H1 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A, and Y407V substitutions, L1 comprises E124C, L135F, and C214S substitutions, and H2 comprises T366W substitutions; (h) H1 comprises F126C, F170V, S183I, V185L, C220S, and T366W substitutions, L1 comprises E124C, L135F, and C214S substitutions, and H2 comprises T366S, L368A, and Y407V substitutions;(i) H1 comprises F126C, C220S, T366S, L368A, and Y407V substitutions, L1 comprises E124C and C214S substitutions, and H2 comprises T366W substitutions; or(j) H1 comprises F126C, C220S, and T366W substitutions, L1 comprises E124C and C214S substitutions, and H2 comprises T366S, L368A, and Y407V substitutions; andwherein the amino acid positions are according to EU numbering.

實施例56.    如實施例53至55中任一者之多特異性構築體，其中該Fc域係衍生自人類IgG1，其中該L1-CL係衍生自人類λ輕鏈，且其中： (a)該H1包含F170I、S183L、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代； (b)該H1包含F170I、S183L、V185L、L234A、L235A、T366W、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (c)該H1包含F170V、S183I、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代； (d)該H1包含F170V、S183I、V185L、L234A、L235A、T366W、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (e)該H1包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代； (f)該H1包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (g)該H1包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代； (h)該H1包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (i)該H1包含F126C、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代；或 (j)該H1包含F126C、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；且 其中該胺基酸位置係根據EU編號。Embodiment 56. The multispecific construct of any one of Embodiments 53 to 55, wherein the Fc domain is derived from human IgG1, wherein the L1-CL is derived from a human lambda light chain, and wherein:(a) the H1 comprises F170I, S183L, V185L, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, the L1 comprises an L135F substitution, and the H2 comprises L234A, L235A, T366W, M428L, and N434S substitutions; (b) H1 comprises F170I, S183L, V185L, L234A, L235A, T366W, M428L, and N434S substitutions, L1 comprises L135F substitutions, and H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions;(c) H1 comprises F170V, S183I, V185L, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, L1 comprises L135F substitutions, and H2 comprises L234A, L235A, T366W, M428L, and N434S substitutions; (d) H1 comprises F170V, S183I, V185L, L234A, L235A, T366W, M428L and N434S substitutions, L1 comprises L135F substitution, and H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (e) H1 comprises F126C, F170I, S183L, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, L1 comprises E124C, L135F and C214S substitutions, and H2 comprises L234A, L235A, T366W, M428L and N434S substitutions; (f) the H1 comprises F126C, F170I, S183L, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (g) H1 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, L1 comprises E124C, L135F and C214S substitutions, and H2 comprises L234A, L235A, T366W, M428L and N434S substitutions; (h) H1 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, L1 comprises E124C, L135F and C214S substitutions, and H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (i) H1 comprises F126C, C220S, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, L1 comprises E124C and C214S substitutions, and H2 comprises L234A, L235A, T366W, M428L, and N434S substitutions; or(j) H1 comprises F126C, C220S, L234A, L235A, T366W, M428L, and N434S substitutions, L1 comprises E124C and C214S substitutions, and H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions; and The amino acid positions are based on EU numbering.

實施例57.    如實施例54至56中任一者之多特異性構築體，其中： (a)該H1含有SEQ ID NO:87之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列； (b)該H1含有SEQ ID NO:88之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列； (c)該H1含有SEQ ID NO:89之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列； (d)該H1含有SEQ ID NO:90之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列； (e)該H1含有SEQ ID NO:91之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列； (f)該H1含有SEQ ID NO:92之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列； (g)該H1含有SEQ ID NO:93之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列； (h)該H1含有SEQ ID NO:94之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列； (i)該H1含有SEQ ID NO:198之胺基酸序列，該L1含有SEQ ID NO:204或205之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列；或 (j)該H1含有SEQ ID NO:199之胺基酸序列，該L1含有SEQ ID NO:204或205之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列。Example 57. The multispecific construct of any one of Examples 54 to 56, wherein:(a) H1 contains the amino acid sequence of SEQ ID NO: 87, L1 contains the amino acid sequence of SEQ ID NO: 173 or 202, and H2 contains the amino acid sequence of SEQ ID NO: 85;(b) H1 contains the amino acid sequence of SEQ ID NO: 88, L1 contains the amino acid sequence of SEQ ID NO: 173 or 202, and H2 contains the amino acid sequence of SEQ ID NO: 86;(c) H1 contains the amino acid sequence of SEQ ID NO: 89, L1 contains the amino acid sequence of SEQ ID NO: 173 or 202, and H2 contains the amino acid sequence of SEQ ID NO: 85;(d) H1 contains the amino acid sequence of SEQ ID NO: 90, L1 contains the amino acid sequence of SEQ ID NO: NO:173 or 202, and H2 contains the amino acid sequence of SEQ ID NO:86;(e) H1 contains the amino acid sequence of SEQ ID NO:91, L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and H2 contains the amino acid sequence of SEQ ID NO:85;(f) H1 contains the amino acid sequence of SEQ ID NO:92, L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and H2 contains the amino acid sequence of SEQ ID NO:86;(g) H1 contains the amino acid sequence of SEQ ID NO:93, L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and H2 contains the amino acid sequence of SEQ ID NO:85;(h) H1 contains the amino acid sequence of SEQ ID NO:94, L1 contains the amino acid sequence of SEQ ID NO: (i) the H1 contains the amino acid sequence of SEQ ID NO: 198, the L1 contains the amino acid sequence of SEQ ID NO: 204 or 205, and the H2 contains the amino acid sequence of SEQ ID NO: 85; or (j) the H1 contains the amino acid sequence of SEQ ID NO: 199, the L1 contains the amino acid sequence of SEQ ID NO: 204 or 205, and the H2 contains the amino acid sequence of SEQ ID NO: 86.

實施例58.    如實施例54至57中任一者之多特異性構築體，其中該L2-CL係衍生自人類κ輕鏈。Embodiment 58. The multispecific construct of any one of embodiments 54 to 57, wherein the L2-CL is derived from a human kappa light chain.

實施例59.    如實施例53至58中任一者之多特異性構築體，其中該第二抗原為TSLP。Embodiment 59. The multispecific construct of any one of Embodiments 53 to 58, wherein the second antigen is TSLP.

實施例60.    如實施例59之多特異性構築體，其中： (a)該H1含有SEQ ID NO:139之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (b)該H1含有SEQ ID NO:138之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (c)該H1含有SEQ ID NO:141之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (d)該H1含有SEQ ID NO:140之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (e)該H1含有SEQ ID NO:126之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (f)該H1含有SEQ ID NO:127之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (g)該H1含有SEQ ID NO:128之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (h)該H1含有SEQ ID NO:129之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (i)該H1含有SEQ ID NO:200之胺基酸序列，該L1含有SEQ ID NO:121或206之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；或 (j)該H1含有SEQ ID NO:201之胺基酸序列，該L1含有SEQ ID NO:121或206之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。Example 60. The multispecific construct of Example 59, wherein:(a) H1 contains the amino acid sequence of SEQ ID NO: 139, L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, H2 contains the amino acid sequence of SEQ ID NO: 136, and L2 contains the amino acid sequence of SEQ ID NO: 103;(b) H1 contains the amino acid sequence of SEQ ID NO: 138, L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, H2 contains the amino acid sequence of SEQ ID NO: 137, and L2 contains the amino acid sequence of SEQ ID NO: 103;(c) H1 contains the amino acid sequence of SEQ ID NO: 141, L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, and H2 contains the amino acid sequence of SEQ ID NO: (d) the H1 contains the amino acid sequence of SEQ ID NO: 140, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (e) the H1 contains the amino acid sequence of SEQ ID NO: 126, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (f) the H1 contains the amino acid sequence of SEQ ID NO: 127, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, and the H2 contains the amino acid sequence of SEQ ID NO: 137. (g) the H1 contains the amino acid sequence of SEQ ID NO: 128, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (h) the H1 contains the amino acid sequence of SEQ ID NO: 129, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (i) the H1 contains the amino acid sequence of SEQ ID NO: 200, the L1 contains the amino acid sequence of SEQ ID NO: 121 or 206, and the H2 contains the amino acid sequence of SEQ ID NO: 137. (j) the H1 contains the amino acid sequence of SEQ ID NO: 201, the L1 contains the amino acid sequence of SEQ ID NO: 121 or 206, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.

實施例61.    如實施例53之多特異性構築體，其中該Fc域係衍生自人類IgG1，其中該L2-CL係衍生自人類λ輕鏈，且其中： (a)該H2包含F170I、S183L及V185L取代，且該L2包含L135F取代； (b)該H2包含F170V、S183I及V185L取代，且該L2包含L135F取代； (c)該H2包含F126C、F170I、S183L、V185L及C220S取代，且該L2包含E124C、L135F及C214S取代； (d)該H2包含F126C、F170V、S183I、V185L及C220S取代，且該L2包含E124C、L135F及C214S取代；或 (e)該H2包含F126C及C220S取代，且該L2包含E124C及C214S取代；且 其中該胺基酸位置係根據EU編號。Example 61. The multispecific construct of Example 53, wherein the Fc domain is derived from human IgG1, wherein the L2-CL is derived from a human lambda light chain, and wherein:(a) the H2 comprises F170I, S183L, and V185L substitutions, and the L2 comprises an L135F substitution;(b) the H2 comprises F170V, S183I, and V185L substitutions, and the L2 comprises an L135F substitution;(c) the H2 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and the L2 comprises E124C, L135F, and C214S substitutions; (d) H2 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and L2 comprises E124C, L135F, and C214S substitutions; or(e) H2 comprises F126C and C220S substitutions, and L2 comprises E124C and C214S substitutions; andwherein the amino acid positions are according to EU numbering.

實施例62.    如實施例53或61之多特異性構築體，其中該Fc域係衍生自人類IgG1，其中該L2-CL係衍生自人類λ輕鏈，其中： (a)該H2包含F170I、S183L、V185L、T366S、L368A及Y407V取代，該L2包含L135F取代，且該H1包含T366W取代； (b)該H2包含F170I、S183L、V185L及T366W取代，該L2包含L135F取代，且該H1包含T366S、L368A及Y407V取代； (c)該H2包含F170V、S183I、V185L、T366S、L368A及Y407V取代，該L2包含L135F取代，且該H1包含T366W取代； (d)該H2包含F170V、S183I、V185L及T366W取代，該L2包含L135F取代，且該H1包含T366S、L368A及Y407V取代； (e)該H2包含F126C、F170I、S183L、V185L、C220S、T366S、L368A及Y407V取代，該L2包含E124C、L135F及C214S取代，且該H1包含T366W取代； (f)該H2包含F126C、F170I、S183L、V185L、C220S及T366W取代，該L2包含E124C、L135F及C214S取代，且該H1包含T366S、L368A及Y407V取代； (g)該H2包含F126C、F170V、S183I、V185L、C220S、T366S、L368A及Y407V取代，該L2包含E124C、L135F及C214S取代，且該H1包含T366W取代； (h)該H2包含F126C、F170V、S183I、V185L、C220S及T366W取代，該L2包含E124C、L135F及C214S取代，且該H1包含T366S、L368A及Y407V取代； (i)該H2包含F126C、C220S、T366S、L368A及Y407V取代，該L2包含E124C及C214S取代，且該H1包含T366W取代；或(j)該H2包含F126C、C220S及T366W取代，該L2包含E124C及C214S取代，且該H1包含T366S、L368A及Y407V取代；且 其中該胺基酸位置係根據EU編號。Example 62. The multispecific construct of Example 53 or 61, wherein the Fc domain is derived from human IgG1, wherein the L2-CL is derived from a human lambda light chain, wherein:(a) H2 comprises F170I, S183L, V185L, T366S, L368A, and Y407V substitutions, L2 comprises L135F substitution, and H1 comprises T366W substitution;(b) H2 comprises F170I, S183L, V185L, and T366W substitutions, L2 comprises L135F substitution, and H1 comprises T366S, L368A, and Y407V substitutions; (c) H2 comprises F170V, S183I, V185L, T366S, L368A, and Y407V substitutions, L2 comprises L135F substitutions, and H1 comprises T366W substitutions;(d) H2 comprises F170V, S183I, V185L, and T366W substitutions, L2 comprises L135F substitutions, and H1 comprises T366S, L368A, and Y407V substitutions;(e) H2 comprises F126C, F170I, S183L, V185L, C220S, T366S, L368A, and Y407V substitutions, L2 comprises E124C, L135F, and C214S substitutions, and H1 comprises T366W substitutions; (f) H2 comprises F126C, F170I, S183L, V185L, C220S, and T366W substitutions, L2 comprises E124C, L135F, and C214S substitutions, and H1 comprises T366S, L368A, and Y407V substitutions;(g) H2 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A, and Y407V substitutions, L2 comprises E124C, L135F, and C214S substitutions, and H1 comprises T366W substitutions; (h) H2 comprises F126C, F170V, S183I, V185L, C220S, and T366W substitutions, L2 comprises E124C, L135F, and C214S substitutions, and H1 comprises T366S, L368A, and Y407V substitutions;(i) H2 comprises F126C, C220S, T366S, L368A, and Y407V substitutions, L2 comprises E124C and C214S substitutions, and H1 comprises T366W substitutions; or (j) H2 comprises F126C, C220S, and T366W substitutions, L2 comprises E124C and C214S substitutions, and H1 comprises T366S, L368A, and Y407V substitutions; andwherein the amino acid positions are according to EU numbering.

實施例63.    如實施例53、61及62中任一者之多特異性構築體，其中該Fc域係衍生自人類IgG1，其中該L2-CL係衍生自人類λ輕鏈，且其中： (a)該H2包含F170I、S183L、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含L135F取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代； (b)該H2包含F170I、S183L、V185L、L234A、L235A、T366W、M428L及N434S取代，該L2包含L135F取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (c)該H2包含F170V、S183I、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含L135F取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代； (d)該H2包含F170V、S183I、V185L、L234A、L235A、T366W、M428L及N434S取代，該L2包含L135F取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (e)該H2包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含E124C、L135F及C214S取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代； (f)該H2包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L2包含E124C、L135F及C214S取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (g)該H2包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含E124C、L135F及C214S取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代； (h)該H2包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L2包含E124C、L135F及C214S取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (i)該H2包含F126C、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L2包含E124C及C214S取代，且該H1包含L234A、L235A、T366W、M428L及N434S取代；或 (j)該H2包含F126C、C220S、L234A、L235A、T366W、M428L及N434S取代，該L2包含E124C及C214S取代，且該H1包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；且 其中該胺基酸位置係根據EU編號。Embodiment 63. The multispecific construct of any one of Embodiments 53, 61, and 62, wherein the Fc domain is derived from human IgG1, wherein the L2-CL is derived from a human lambda light chain, and wherein:(a) the H2 comprises F170I, S183L, V185L, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, the L2 comprises an L135F substitution, and the H1 comprises L234A, L235A, T366W, M428L, and N434S substitutions; (b) H2 comprises F170I, S183L, V185L, L234A, L235A, T366W, M428L, and N434S substitutions, L2 comprises L135F substitutions, and H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions;(c) H2 comprises F170V, S183I, V185L, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, L2 comprises L135F substitutions, and H1 comprises L234A, L235A, T366W, M428L, and N434S substitutions; (d) H2 comprises F170V, S183I, V185L, L234A, L235A, T366W, M428L and N434S substitutions, L2 comprises L135F substitution, and H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (e) H2 comprises F126C, F170I, S183L, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, L2 comprises E124C, L135F and C214S substitutions, and H1 comprises L234A, L235A, T366W, M428L and N434S substitutions; (f) H2 comprises F126C, F170I, S183L, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, L2 comprises E124C, L135F and C214S substitutions, and H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (g) H2 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, L2 comprises E124C, L135F and C214S substitutions, and H1 comprises L234A, L235A, T366W, M428L and N434S substitutions; (h) H2 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, L2 comprises E124C, L135F and C214S substitutions, and H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (i) H2 comprises F126C, C220S, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, L2 comprises E124C and C214S substitutions, and H1 comprises L234A, L235A, T366W, M428L, and N434S substitutions; or(j) H2 comprises F126C, C220S, L234A, L235A, T366W, M428L, and N434S substitutions, L2 comprises E124C and C214S substitutions, and H1 comprises L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions; and The amino acid positions are based on EU numbering.

實施例64.    如實施例61至63中任一者之多特異性構築體，其中： (a)該H2含有SEQ ID NO:87之胺基酸序列，該L2含有SEQ ID NO:173或202之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列； (b)該H2含有SEQ ID NO:88之胺基酸序列，該L2含有SEQ ID NO:173或202之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列； (c)該H2含有SEQ ID NO:89之胺基酸序列，該L2含有SEQ ID NO:173或202之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列； (d)該H2含有SEQ ID NO:90之胺基酸序列，該L2含有SEQ ID NO:173或202之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列； (e)該H2含有SEQ ID NO:91之胺基酸序列，該L2含有SEQ ID NO:174或203之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列； (f)該H2含有SEQ ID NO:92之胺基酸序列，該L2含有SEQ ID NO:174或203之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列； (g)該H2含有SEQ ID NO:93之胺基酸序列，該L2含有SEQ ID NO:174或203之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列； (h)該H2含有SEQ ID NO:94之胺基酸序列，該L2含有SEQ ID NO:174或203之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列； (i)該H2含有SEQ ID NO:198之胺基酸序列，該L2含有SEQ ID NO:204或205之胺基酸序列，且該H1含有SEQ ID NO:85之胺基酸序列；或 (j)該H2含有SEQ ID NO:199之胺基酸序列，該L2含有SEQ ID NO:204或205之胺基酸序列，且該H1含有SEQ ID NO:86之胺基酸序列。Example 64. The multispecific construct of any one of Examples 61 to 63, wherein:(a) H2 contains the amino acid sequence of SEQ ID NO:87, L2 contains the amino acid sequence of SEQ ID NO:173 or 202, and H1 contains the amino acid sequence of SEQ ID NO:85;(b) H2 contains the amino acid sequence of SEQ ID NO:88, L2 contains the amino acid sequence of SEQ ID NO:173 or 202, and H1 contains the amino acid sequence of SEQ ID NO:86;(c) H2 contains the amino acid sequence of SEQ ID NO:89, L2 contains the amino acid sequence of SEQ ID NO:173 or 202, and H1 contains the amino acid sequence of SEQ ID NO:85;(d) H2 contains the amino acid sequence of SEQ ID NO:90, L2 contains the amino acid sequence of SEQ ID NO: (e) the H2 contains the amino acid sequence of SEQ ID NO:91, the L2 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H1 contains the amino acid sequence of SEQ ID NO:85; (f) the H2 contains the amino acid sequence of SEQ ID NO:92, the L2 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H1 contains the amino acid sequence of SEQ ID NO:86; (g) the H2 contains the amino acid sequence of SEQ ID NO:93, the L2 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H1 contains the amino acid sequence of SEQ ID NO:85; (h) the H2 contains the amino acid sequence of SEQ ID NO:94, the L2 contains the amino acid sequence of SEQ ID NO: (i) the H2 contains the amino acid sequence of SEQ ID NO: 198, the L2 contains the amino acid sequence of SEQ ID NO: 204 or 205, and the H1 contains the amino acid sequence of SEQ ID NO: 85; or (j) the H2 contains the amino acid sequence of SEQ ID NO: 199, the L2 contains the amino acid sequence of SEQ ID NO: 204 or 205, and the H1 contains the amino acid sequence of SEQ ID NO: 86.

實施例65.    如實施例61至64中任一者之多特異性構築體，其中該L1-CL係衍生自人類κ輕鏈。Embodiment 65. The multispecific construct of any one of embodiments 61 to 64, wherein the L1-CL is derived from a human kappa light chain.

實施例66.    如實施例61至65中任一者之多特異性構築體，其中該第二抗原為TSLP。Example 66. The multispecific construct of any one of Examples 61 to 65, wherein the second antigen is TSLP.

實施例67.    如實施例1至52中任一者之多特異性構築體，其中該特異性地結合至IL-13的第一抗體部分與該特異性地結合至第二標靶抗原的第二抗體部分係經由連接子彼此融合。Embodiment 67. The multispecific construct of any one of Embodiments 1 to 52, wherein the first antibody portion that specifically binds to IL-13 and the second antibody portion that specifically binds to a second target antigen are fused to each other via a linker.

實施例68.    如實施例67之多特異性構築體，其中該連接子含有GG及SEQ ID NO:98-99之任一者的胺基酸序列。Example 68. The multispecific construct of Example 67, wherein the linker contains GG and an amino acid sequence of any one of SEQ ID NOs: 98-99.

實施例69.    如實施例1至10、17至23、67及68中任一者之多特異性構築體，其中該第一抗體部分為scFv (「抗IL-13 scFv」)，其中該第二抗體部分特異性地結合至第二標靶抗原的全長抗體，且其中該抗IL-13 scFv係融合至第二抗體部分之重鏈之一者的C端以形成融合多肽。Embodiment 69. The multispecific construct of any one of Embodiments 1 to 10, 17 to 23, 67 and 68, wherein the first antibody portion is a scFv ("anti-IL-13 scFv"), wherein the second antibody portion specifically binds to a full-length antibody of a second target antigen, and wherein the anti-IL-13 scFv is fused to the C-terminus of one of the heavy chains of the second antibody portion to form a fusion polypeptide.

實施例70.    如實施例1至10、17至23及67至69中任一者之多特異性構築體，其中該多特異性構築體包含兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)，以及第二抗體部分，其為特異性地結合至第二標靶抗原的全長抗體；且其中抗IL-13 scFv1係融合至第二抗體部分之第一重鏈的C端以形成第一融合多肽，且抗IL-13 scFv2係融合至第二抗體部分之第二重鏈的C端以形成第二融合多肽。Embodiment 70. The multispecific construct of any one of Embodiments 1 to 10, 17 to 23, and 67 to 69, wherein the multispecific construct comprises two anti-IL-13 antibody portions, which are scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen; and wherein the anti-IL-13 scFv1 is fused to the C-terminus of the first heavy chain of the second antibody portion to form a first fusion polypeptide, and the anti-IL-13 scFv2 is fused to the C-terminus of the second heavy chain of the second antibody portion to form a second fusion polypeptide.

實施例71.    如實施例70之多特異性構築體，其中該第二標靶抗原為TSLP。Example 71. The multispecific construct of Example 70, wherein the second target antigen is TSLP.

實施例72.    如實施例71之多特異性構築體，其中該第二抗體部分為抗TSLP全長抗體，其包含兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈及兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:113之胺基酸序列。Example 72. The multispecific construct of Example 71, wherein the second antibody portion is an anti-TSLP full-length antibody comprising two light chains each containing the amino acid sequence of SEQ ID NO: 103 and two heavy chains each containing the amino acid sequence of SEQ ID NO: 105, and wherein the first fusion polypeptide and the second fusion polypeptide each contain the amino acid sequence of SEQ ID NO: 113.

實施例73.    如實施例中任一者之多特異性構築體1至10、17至19、26、27、67及68，其中該多特異性構築體包含兩個抗IL-13抗體部分，其等為scFv (「抗IL-13 scFv1」及「抗IL-13 scFv2」)、兩個第二抗體部分，其等為特異性地結合至第二標靶抗原的Fab (「Fab1」及「Fab2」)，以及Fc域； 其中該多特異性構築體包含： i)含有N’至C’：VL1-(L1-CL)的第一多肽； ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-抗IL-13 scFv1-視情況的連接子-Fc域之第一次單元的第二多肽； iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-抗IL-13 scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及 iv)含有N’至C’：VL2-(L2-CL)的第四多肽；且 其中VL1-(L1-CL)及VH1-(H1-CH1)形成Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成Fab2。Embodiment 73. The multispecific construct of any of Embodiments 1 to 10, 17 to 19, 26, 27, 67, and 68, wherein the multispecific construct comprises two anti-IL-13 antibody portions, which are scFvs ("anti-IL-13 scFv1" and "anti-IL-13 scFv2"), two second antibody portions, which are Fabs that specifically bind to a second target antigen ("Fab1" and "Fab2"), and an Fc domain; wherein the multispecific construct comprises: i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a first polypeptide comprising N' to C': VH1-(H1-CH1)-optionally a linker-anti-IL-13 scFv1-optional linker-Fc domain;iii) a third polypeptide comprising N' to C': VH2-(H2-CH1)-optional linker-anti-IL-13 scFv2-optional linker-Fc domain; andiv) a fourth polypeptide comprising N' to C': VL2-(L2-CL); andwherein VL1-(L1-CL) and VH1-(H1-CH1) form Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form Fab2.

實施例74.    如實施例73之多特異性構築體，其中該第二標靶抗原為TSLP。Example 74. The multispecific construct of Example 73, wherein the second target antigen is TSLP.

實施例75.    如實施例74之多特異性構築體，其中該第一多肽及該第四多肽各自含有SEQ ID NO:103之胺基酸序列，且該第二多肽及該第三多肽各自含有SEQ ID NO:110之胺基酸序列。Example 75. The multispecific construct of Example 74, wherein the first polypeptide and the fourth polypeptide each contain the amino acid sequence of SEQ ID NO: 103, and the second polypeptide and the third polypeptide each contain the amino acid sequence of SEQ ID NO: 110.

實施例76.    如實施例中任一者之多特異性構築體1至8、13至19、24、25、28至52、67及68，其中該第一抗體部分為全長抗體(「抗IL-13全長抗體」)，其中該第二抗體部分為特異性地結合至該第二標靶抗原的scFv，且其中該scFv係融合至該抗IL-13全長抗體之重鏈之一者的C端以形成融合多肽。Embodiment 76. The multispecific construct of any one of Embodiments 1 to 8, 13 to 19, 24, 25, 28 to 52, 67 and 68, wherein the first antibody portion is a full-length antibody ("anti-IL-13 full-length antibody"), wherein the second antibody portion is a scFv that specifically binds to the second target antigen, and wherein the scFv is fused to the C-terminus of one of the heavy chains of the anti-IL-13 full-length antibody to form a fusion polypeptide.

實施例77.    如實施例1至8、13至19、24、25、28至52、67、68及76中任一者之多特異性構築體，其中該多特異性構築體包含該第一抗體部分，其為抗IL-13全長抗體，以及兩個第二抗體部分，其等為scFv (「scFv1」及「scFv2」) 特異性地結合至第二標靶抗原；且其中scFv1係融合至該抗IL-13全長抗體之第一重鏈的C端以形成第一融合多肽，且scFv2係融合至該抗IL-13全長抗體之第二重鏈的C端以形成第二融合多肽。Embodiment 77. The multispecific construct of any one of Embodiments 1 to 8, 13 to 19, 24, 25, 28 to 52, 67, 68 and 76, wherein the multispecific construct comprises the first antibody portion, which is an anti-IL-13 full-length antibody, and two second antibody portions, which are scFvs ("scFv1" and "scFv2") that specifically bind to a second target antigen; and wherein scFv1 is fused to the C-terminus of the first heavy chain of the anti-IL-13 full-length antibody to form a first fusion polypeptide, and scFv2 is fused to the C-terminus of the second heavy chain of the anti-IL-13 full-length antibody to form a second fusion polypeptide.

實施例78.    如實施例77之多特異性構築體，其中該第二標靶抗原為TSLP。Example 78. The multispecific construct of Example 77, wherein the second target antigen is TSLP.

實施例79.    如實施例78之多特異性構築體， (i)其中該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:111之胺基酸序列；或 (ii)其中該抗IL-13全長抗體包含兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈，且其中該第一融合多肽及該第二融合多肽各自含有SEQ ID NO:220-223之任一者的胺基酸序列。Example 79. The multispecific construct of Example 78, wherein:(i) the anti-IL-13 full-length antibody comprises two heavy chains each comprising the amino acid sequence of SEQ ID NO: 125 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197, and wherein the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of SEQ ID NO: 111; or(ii) the anti-IL-13 full-length antibody comprises two heavy chains each comprising the amino acid sequence of SEQ ID NO: 225 and two light chains each comprising the amino acid sequence of SEQ ID NO: 102 or 197, and wherein the first fusion polypeptide and the second fusion polypeptide each comprise the amino acid sequence of any one of SEQ ID NOs: 220-223.

實施例80.    如實施例1至8、11、12、17至19、24、25、28至52、67及68中任一者之多特異性構築體，其中該多特異性構築體包含兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)、兩個第二抗體部分，其等為特異性地結合至第二標靶抗原的scFv (「scFv1」及「scFv2」)，以及Fc域； 其中該多特異性構築體包含： i)含有N’至C’：VL1-(L1-CL)的第一多肽； ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-scFv1-視情況的連接子-Fc域之第一次單元的第二多肽； iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-scFv2-視情況的連接子-Fc域之第二次單元的第三多肽；及 iv)含有N’至C’：VL2-(L2-CL)的第四多肽；且 其中VH1-(H1-CH1)及VL1-(L1-CL)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2。Embodiment 80. The multispecific construct of any one of Embodiments 1 to 8, 11, 12, 17 to 19, 24, 25, 28 to 52, 67, and 68, wherein the multispecific construct comprises two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2"), two second antibody portions, which are scFvs that specifically bind to a second target antigen ("scFv1" and "scFv2"), and an Fc domain; wherein the multispecific construct comprises:i) a first polypeptide comprising N' to C': VL1-(L1-CL); ii) a second polypeptide comprising a first unit comprising N’ to C’: VH1-(H1-CH1)-optional linker-scFv1-optional linker-Fc domain;iii) a third polypeptide comprising a second unit comprising N’ to C’: VH2-(H2-CH1)-optional linker-scFv2-optional linker-Fc domain; andiv) a fourth polypeptide comprising N’ to C’: VL2-(L2-CL); andwherein VH1-(H1-CH1) and VL1-(L1-CL) form an anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form an anti-IL-13 Fab2.

實施例81.    如實施例80之多特異性構築體，其中該第二標靶抗原為TSLP。Example 81. The multispecific construct of Example 80, wherein the second target antigen is TSLP.

實施例82.    如實施例81之多特異性構築體，其中該第一多肽及該第四多肽各自含有SEQ ID NO:102或197之胺基酸序列，且該第二多肽及該第三多肽各自含有SEQ ID NO:112之胺基酸序列。Example 82. The multispecific construct of Example 81, wherein the first polypeptide and the fourth polypeptide each contain the amino acid sequence of SEQ ID NO: 102 or 197, and the second polypeptide and the third polypeptide each contain the amino acid sequence of SEQ ID NO: 112.

實施例83.    如實施例1至8、11、12、17至23、28至41、67及68中任一者之多特異性構築體，其中該多特異性包含兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)，以及第二抗體部分，其為特異性地結合至第二標靶抗原的全長抗體； 其中該多特異性構築體包含： i)含有第二抗體部分之第一輕鏈的第一多肽； ii)含有N’至C’：第二抗體部分之第一重鏈-視情況的連接子-VH1-(H1-CH1)的第二多肽； iii)含有N’至C’：第二抗體部分之第二重鏈-視情況的連接子-VH2-(H2-CH1)的第三多肽； iv)含有第二抗體部分之第二輕鏈的第四多肽； v)含有N’至C’：VL1-(L1-CL)的第五多肽；及 vi)含有N’至C’：VL2-(L2-CL)的第六多肽；且 其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VL2-(L2-CL)及VH2-(H2-CH1)形成抗IL-13 Fab2。Embodiment 83. A multispecific construct according to any one of Embodiments 1 to 8, 11, 12, 17 to 23, 28 to 41, 67, and 68, wherein the multispecific construct comprises two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2"), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen; wherein the multispecific construct comprises:i) a first polypeptide comprising the first light chain of the second antibody portion;ii) a second polypeptide comprising the first heavy chain of the second antibody portion (N' to C'): VH1-(H1-CH1)-optionally a linker; iii) a third polypeptide comprising N’ to C’: the second heavy chain of the second antibody moiety, optionally a linker, VH2-(H2-CH1);iv) a fourth polypeptide comprising the second light chain of the second antibody moiety;v) a fifth polypeptide comprising N’ to C’: VL1-(L1-CL); andvi) a sixth polypeptide comprising N’ to C’: VL2-(L2-CL); andwherein VL1-(L1-CL) and VH1-(H1-CH1) form anti-IL-13 Fab1, and VL2-(L2-CL) and VH2-(H2-CH1) form anti-IL-13 Fab2.

實施例84.    如實施例83之多特異性構築體，其中該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中： (a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代； (b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代； (c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代； (d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；或 (e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C及C214S取代；且 其中該胺基酸位置係根據EU編號。Example 84. The multispecific construct of Example 83, wherein the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein:(a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions;(b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions;(c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions;(d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; or(e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C and C214S substitutions; andwherein the amino acid positions are according to EU numbering.

實施例85.    如實施例83或84之多特異性構築體，其中該第二標靶抗原為TSLP。Example 85. The multispecific construct of Example 83 or 84, wherein the second target antigen is TSLP.

實施例86.    如實施例85之多特異性構築體，其中該第二抗體部分為抗TSLP全長抗體，其包含兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈。Example 86. The multispecific construct of Example 85, wherein the second antibody portion is an anti-TSLP full-length antibody comprising two heavy chains each containing the amino acid sequence of SEQ ID NO: 105 and two light chains each containing the amino acid sequence of SEQ ID NO: 103.

實施例87.    如實施例1至8、13至19、26至52、67及68中任一者之多特異性構築體，其中該多特異性構築體包含抗IL-13抗體部分，其為全長抗體(「抗IL-13全長抗體」)，以及兩個第二抗體部分，其等為Fab (「Fab1」及「Fab2」) 特異性地結合至第二標靶抗原； 其中該多特異性構築體包含： i)含有抗IL-13全長抗體之第一輕鏈(L1)的第一多肽； ii)含有N’至C’：抗IL-13全長抗體之第一重鏈(H1)-視情況的連接子-VH3-(H3-CH1)的第二多肽； iii)含有N’至C’：抗IL-13全長抗體之第二重鏈(H2)-視情況的連接子-VH4-(H4-CH1)的第三多肽； iv)含有抗IL-13全長抗體之第二輕鏈(L2)的第四多肽； v)含有N’至C’：VL3-(L3-CL)的第五多肽；及 vi)含有N’至C’：VL4-(L4-CL)的第六多肽；且 其中VH3-(H3-CH1)及VL3-(L3-CL)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2。Embodiment 87. A multispecific construct according to any one of Embodiments 1 to 8, 13 to 19, 26 to 52, 67, and 68, wherein the multispecific construct comprises an anti-IL-13 antibody portion that is a full-length antibody ("anti-IL-13 full-length antibody") and two second antibody portions that are Fabs ("Fab1" and "Fab2") that specifically bind to a second target antigen; wherein the multispecific construct comprises:i) a first polypeptide comprising the first light chain (L1) of the anti-IL-13 full-length antibody;ii) a second polypeptide comprising N' to C': the first heavy chain (H1) of the anti-IL-13 full-length antibody - optionally a linker - VH3 - (H3-CH1); iii) a third polypeptide comprising N’ to C’: the second heavy chain (H2) of the anti-IL-13 full-length antibody, optionally a linker, and VH4-(H4-CH1);iv) a fourth polypeptide comprising the second light chain (L2) of the anti-IL-13 full-length antibody;v) a fifth polypeptide comprising N’ to C’: VL3-(L3-CL); andvi) a sixth polypeptide comprising N’ to C’: VL4-(L4-CL); andwherein VH3-(H3-CH1) and VL3-(L3-CL) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2.

實施例88.    如實施例87之多特異性構築體，其中： (a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代； (b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代； (c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代； (d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；或 (e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C及C214S取代；且 其中該胺基酸位置係根據EU編號。Example 88. The multispecific construct of Example 87, wherein:(a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions;(b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions;(c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions;(d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; or(e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C and C214S substitutions; andwherein the amino acid positions are according to EU numbering.

實施例89.    如實施例87或88之多特異性構築體，其中該抗IL-13全長抗體包含： i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； ii)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； iii)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； iv)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈； v)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； vi)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； vii)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； viii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或 ix)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈。Example 89. The multispecific construct of Example 87 or 88, wherein the full-length anti-IL-13 antibody comprises:i) two heavy chains each containing the amino acid sequence of SEQ ID NO: 125 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197;ii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 209 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207;iii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 210 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207;iv) two heavy chains each containing the amino acid sequence of SEQ ID NO: 211 and two light chains each containing the amino acid sequence of SEQ ID NO: NO:121 or 206;v) two heavy chains each containing the amino acid sequence of SEQ ID NO:212 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208;vi) two heavy chains each containing the amino acid sequence of SEQ ID NO:213 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208;vii) two heavy chains each containing the amino acid sequence of SEQ ID NO:225 and two light chains each containing the amino acid sequence of SEQ ID NO:102 or 197;viii) two heavy chains each containing the amino acid sequence of SEQ ID NO:101 and two light chains each containing the amino acid sequence of SEQ ID NO:102 or 197; orix) two heavy chains each containing the amino acid sequence of SEQ ID NO: A heavy chain containing the amino acid sequence of SEQ ID NO: 123 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206.

實施例90.    如實施例87至89中任一者之多特異性構築體，其中該第二標靶抗原為TSLP (「抗TSLP Fab1」及「抗TSLP Fab2」)。Embodiment 90. The multispecific construct of any one of Embodiments 87 to 89, wherein the second target antigen is TSLP ("anti-TSLP Fab1" and "anti-TSLP Fab2").

實施例91.    如實施例90之多特異性構築體，其中該抗TSLP Fab1及該抗TSLP Fab2各自包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。Example 91. The multispecific construct of Example 90, wherein the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103.

實施例92.    如實施例1至8、11、12、17至23、28至41、67及68中任一者之多特異性構築體，其中該多特異性構築體包含兩個抗IL-13抗體部分，其等為Fab (「抗IL-13 Fab1」及「抗IL-13 Fab2」)，以及第二抗體部分，其為特異性地結合至第二標靶抗原的全長抗體； 其中該多特異性構築體包含： i)含有N’至C’：VL1-(L1-CL)的第一多肽； ii)含有N’至C’：VH1-(H1-CH1)-視情況的連接子-第二抗體部分之第一重鏈的第二多肽； iii)含有N’至C’：VH2-(H2-CH1)-視情況的連接子-第二抗體部分之第二重鏈的第三多肽； iv)含有N’至C’：VL2-(L2-CL)的第四多肽； v)含有第二抗體部分之第一輕鏈的第五多肽；及 vi)含有第二抗體部分之第二輕鏈的第六多肽；且 其中VL1-(L1-CL)及VH1-(H1-CH1)形成抗IL-13 Fab1，且VH2-(H2-CH1)及VL2-(L2-CL)形成抗IL-13 Fab2。Embodiment 92. A multispecific construct according to any one of Embodiments 1 to 8, 11, 12, 17 to 23, 28 to 41, 67, and 68, wherein the multispecific construct comprises two anti-IL-13 antibody portions, which are Fabs ("anti-IL-13 Fab1" and "anti-IL-13 Fab2"), and a second antibody portion, which is a full-length antibody that specifically binds to a second target antigen; wherein the multispecific construct comprises:i) a first polypeptide comprising N' to C': VL1-(L1-CL);ii) a second polypeptide comprising N' to C': VH1-(H1-CH1) - optionally a linker - a first heavy chain of the second antibody portion; iii) a third polypeptide comprising N’ to C’: VH2-(H2-CH1) - optionally a linker - the second heavy chain of the second antibody moiety;iv) a fourth polypeptide comprising N’ to C’: VL2-(L2-CL);v) a fifth polypeptide comprising the first light chain of the second antibody moiety; andvi) a sixth polypeptide comprising the second light chain of the second antibody moiety; andwherein VL1-(L1-CL) and VH1-(H1-CH1) form anti-IL-13 Fab1, and VH2-(H2-CH1) and VL2-(L2-CL) form anti-IL-13 Fab2.

實施例93.    如實施例92之多特異性構築體，其中該抗IL-13 Fab1包含含有VH1-(H1-CH1)的H1及含有VL1-(L1-CL)的L1；其中該抗IL-13 Fab2包含含有VH2-(H2-CH1)的H2及含有VL2-(L2-CL)的L2；且其中： (a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代； (b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代； (c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代； (d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；或 (e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C及C214S取代；且 其中該胺基酸位置係根據EU編號。Example 93. The multispecific construct of Example 92, wherein the anti-IL-13 Fab1 comprises H1 comprising VH1-(H1-CH1) and L1 comprising VL1-(L1-CL); wherein the anti-IL-13 Fab2 comprises H2 comprising VH2-(H2-CH1) and L2 comprising VL2-(L2-CL); and wherein:(a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions;(b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions;(c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions;(d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; or(e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C and C214S substitutions; andwherein the amino acid positions are according to EU numbering.

實施例94.    如實施例92或93之多特異性構築體，其中該第二標靶抗原為TSLP。Example 94. The multispecific construct of Example 92 or 93, wherein the second target antigen is TSLP.

實施例95.    如實施例94之多特異性構築體，其中該第二抗體部分為抗TSLP全長抗體，其包含兩條各自含有SEQ ID NO:105之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:103之胺基酸序列的輕鏈。Example 95. The multispecific construct of Example 94, wherein the second antibody portion is an anti-TSLP full-length antibody comprising two heavy chains each containing the amino acid sequence of SEQ ID NO: 105 and two light chains each containing the amino acid sequence of SEQ ID NO: 103.

實施例96.    如實施例1至8、13至19、26至52、67及68中任一者之多特異性構築體，其中該多特異性構築體包含該第一抗體部分，其為抗IL-13全長抗體，以及兩個第二抗體部分，其等為特異性地結合至第二標靶抗原的Fab (「Fab1」及「Fab2」)； 其中該多特異性構築體包含： i)含有N’至C’：VL3-(L3-CL)的第一多肽； ii)含有N’至C’：VH3-(H3-CH1)-視情況的連接子-抗IL-13全長抗體之第一重鏈(H1)的第二多肽； iii)含有N’至C’：VH4-(H4-CH1)-視情況的連接子-抗IL-13全長抗體之第二重鏈(H2)的第三多肽； iv)含有N’至C’：VL4-(L4-CL)的第四多肽； v)含有抗IL-13全長抗體之第一輕鏈(L1)的第五多肽；及 vi)含有抗IL-13全長抗體之第二輕鏈(L2)的第六多肽；且 其中VL3-(L3-CL)及VH3-(H3-CH1)形成Fab1，且VH4-(H4-CH1)及VL4-(L4-CL)形成Fab2。Embodiment 96. The multispecific construct of any one of Embodiments 1 to 8, 13 to 19, 26 to 52, 67, and 68, wherein the multispecific construct comprises the first antibody portion, which is a full-length anti-IL-13 antibody, and two second antibody portions, which are Fabs ("Fab1" and "Fab2") that specifically bind to a second target antigen; wherein the multispecific construct comprises:i) a first polypeptide comprising N' to C': VL3-(L3-CL);ii) a second polypeptide comprising N' to C': VH3-(H3-CH1) - optionally a linker - the first heavy chain (H1) of the full-length anti-IL-13 antibody; iii) a third polypeptide comprising N’ to C’: VH4-(H4-CH1) - optionally a linker - the second heavy chain (H2) of the anti-IL-13 full-length antibody;iv) a fourth polypeptide comprising N’ to C’: VL4-(L4-CL);v) a fifth polypeptide comprising the first light chain (L1) of the anti-IL-13 full-length antibody; andvi) a sixth polypeptide comprising the second light chain (L2) of the anti-IL-13 full-length antibody; andwherein VL3-(L3-CL) and VH3-(H3-CH1) form Fab1, and VH4-(H4-CH1) and VL4-(L4-CL) form Fab2.

實施例97.    如實施例96之多特異性構築體，其中： (a) H1及H2各自包含F170I、S183L及V185L取代，且L1及L2各自包含L135F取代； (b) H1及H2各自包含F170V、S183I及V185L取代，且L1及L2各自包含L135F取代； (c) H1及H2各自包含F126C、F170I、S183L、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代； (d) H1及H2各自包含F126C、F170V、S183I、V185L及C220S取代，且L1及L2各自包含E124C、L135F及C214S取代；或 (e) H1及H2各自包含F126C及C220S取代，且L1及L2各自包含E124C及C214S取代；且 其中該胺基酸位置係根據EU編號。Example 97. The multispecific construct of Example 96, wherein:(a) H1 and H2 each comprise F170I, S183L, and V185L substitutions, and L1 and L2 each comprise L135F substitutions;(b) H1 and H2 each comprise F170V, S183I, and V185L substitutions, and L1 and L2 each comprise L135F substitutions;(c) H1 and H2 each comprise F126C, F170I, S183L, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions;(d) H1 and H2 each comprise F126C, F170V, S183I, V185L, and C220S substitutions, and L1 and L2 each comprise E124C, L135F, and C214S substitutions; or(e) H1 and H2 each comprise F126C and C220S substitutions, and L1 and L2 each comprise E124C and C214S substitutions; andwherein the amino acid positions are according to EU numbering.

實施例98.    如實施例96或97之多特異性構築體，其中該抗IL-13全長抗體包含： i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； ii)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； iii)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； iv)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈； v)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； vi)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； vii)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； viii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或 ix)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈。Example 98. The multispecific construct of Example 96 or 97, wherein the full-length anti-IL-13 antibody comprises:i) two heavy chains each containing the amino acid sequence of SEQ ID NO: 125 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197;ii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 209 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207;iii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 210 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207;iv) two heavy chains each containing the amino acid sequence of SEQ ID NO: 211 and two light chains each containing the amino acid sequence of SEQ ID NO: NO:121 or 206;v) two heavy chains each containing the amino acid sequence of SEQ ID NO:212 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208;vi) two heavy chains each containing the amino acid sequence of SEQ ID NO:213 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208;vii) two heavy chains each containing the amino acid sequence of SEQ ID NO:225 and two light chains each containing the amino acid sequence of SEQ ID NO:102 or 197;viii) two heavy chains each containing the amino acid sequence of SEQ ID NO:101 and two light chains each containing the amino acid sequence of SEQ ID NO:102 or 197; orix) two heavy chains each containing the amino acid sequence of SEQ ID NO: A heavy chain containing the amino acid sequence of SEQ ID NO: 123 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206.

實施例99.    如實施例96至98中任一者之多特異性構築體，其中該第二標靶抗原為TSLP (「抗TSLP Fab1」及「抗TSLP Fab2」)。Embodiment 99. The multispecific construct of any one of Embodiments 96 to 98, wherein the second target antigen is TSLP ("anti-TSLP Fab1" and "anti-TSLP Fab2").

實施例100.  如實施例99之多特異性構築體，其中該抗TSLP Fab1及該抗TSLP Fab2各自包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。Example 100. A multispecific construct as described in Example 99, wherein the anti-TSLP Fab1 and the anti-TSLP Fab2 each comprise a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103.

實施例101.  一種醫藥組成物，其包含如實施例1至100中任一者之多特異性構築體及醫藥上可接受之載體。Example 101. A pharmaceutical composition comprising a multispecific construct according to any one of Examples 1 to 100 and a pharmaceutically acceptable carrier.

實施例102.  一種分離的核酸，其編碼如實施例1至100中任一者之多特異性構築體。Embodiment 102. An isolated nucleic acid encoding a multispecific construct of any one of embodiments 1 to 100.

實施例103.  一種載體，其包含如實施例102之分離的核酸。Example 103. A vector comprising the isolated nucleic acid of Example 102.

實施例104.  一種宿主細胞，其包含如實施例102之分離的核酸或如實施例103之載體。Example 104. A host cell comprising the isolated nucleic acid of Example 102 or the vector of Example 103.

實施例105.  一種治療個體之發炎性疾病的方法，其包含向該個體投予有效量之如實施例1至100中任一者之多特異性構築體或如實施例101之醫藥組成物。Example 105. A method for treating an inflammatory disease in an individual, comprising administering to the individual an effective amount of a multispecific construct of any one of Examples 1 to 100 or a pharmaceutical composition of Example 101.

實施例106.  如實施例105之方法，其中該發炎性疾病為氣喘、異位性皮膚炎或慢性阻塞性肺病(COPD)。Example 106. The method of Example 105, wherein the inflammatory disease is asthma, atopic dermatitis, or chronic obstructive pulmonary disease (COPD).

實施例107.  如實施例105或106之方法，其中該個體為人類。Embodiment 107. The method of embodiment 105 or 106, wherein the individual is a human.

實施例108.  一種產生多特異性構築體之方法，其包含： i)在適合表現該多特異性構築體的條件下培養含有如實施例102之分離的核酸或如實施例103之載體的宿主細胞或如實施例104之宿主細胞；及 ii)獲得該表現的多特異性構築體。Example 108. A method for producing a multispecific construct, comprising:i) culturing a host cell containing the isolated nucleic acid of Example 102 or the vector of Example 103, or the host cell of Example 104, under conditions suitable for expression of the multispecific construct; andii) obtaining the expressed multispecific construct.

實施例109.  一種分離的抗體構築體(「抗IL-13抗體構築體」)，其包含特異性地結合介白素-13的抗體部分(IL-13；「抗IL-13抗體部分」)，其中該抗IL-13抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，且其中：該VH包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多3個胺基酸變異的變體；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多3個胺基酸變異的變體；及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多3個胺基酸變異的變體；且該VL包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1或其含有至多3個胺基酸變異的變體；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多3個胺基酸變異的變體；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多3個胺基酸變異的變體。Example 109. An isolated antibody construct ("anti-IL-13 antibody construct") comprising an antibody portion that specifically binds interleukin-13 (IL-13; "anti-IL-13 antibody portion"), wherein the anti-IL-13 antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), and wherein: the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to 3 amino acid variations; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to 3 amino acid variations; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to 3 amino acid variations; and the VL comprises (i) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to 3 amino acid variations; (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to 3 amino acid variants; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to 3 amino acid variants; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72, or a variant thereof containing up to 3 amino acid variants.

實施例110.  如實施例109之分離的抗IL-13抗體構築體，其中該VH包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3。Example 110. An isolated anti-IL-13 antibody construct as described in Example 109, wherein the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72.

實施例111.  如實施例109或110之分離的抗IL-13抗體構築體，其中該VH含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80%序列同一性的變體，且該VL含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80%序列同一性的變體。Example 111. An isolated anti-IL-13 antibody construct as described in Example 109 or 110, wherein the VH comprises the amino acid sequence of SEQ ID NO:65 or a variant thereof having at least about 80% sequence identity with SEQ ID NO:65, and the VL comprises the amino acid sequence of SEQ ID NO:69 or a variant thereof having at least about 80% sequence identity with SEQ ID NO:69.

實施例112.   如實施例109至111中任一項之分離的抗IL-13抗體構築體，其中該VH含有SEQ ID NO:65之胺基酸序列，且該VL含有SEQ ID NO:69之胺基酸序列。Example 112. An isolated anti-IL-13 antibody construct as described in any one of Examples 109 to 111, wherein the VH contains the amino acid sequence of SEQ ID NO:65 and the VL contains the amino acid sequence of SEQ ID NO:69.

實施例113.  如實施例109至112中任一項之分離的抗IL-13抗體構築體，其中該抗IL-13抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、雙抗體及scFv。Embodiment 113. The isolated anti-IL-13 antibody construct of any one of embodiments 109 to 112, wherein the anti-IL-13 antibody portion is selected from the group consisting of a full-length antibody, Fab, Fab', F(ab')2 , a diabody, and a scFv.

實施例114.  如實施例113之分離的抗IL-13抗體構築體，其中該抗IL-13抗體部分為scFv (「抗IL-13 scFv」)。Example 114. An isolated anti-IL-13 antibody construct as described in Example 113, wherein the anti-IL-13 antibody portion is a scFv ("anti-IL-13 scFv").

實施例115.  如實施例114之分離的抗IL-13抗體構築體，其中該抗IL-13 scFv含有SEQ ID NO:108之胺基酸序列。Example 115. An isolated anti-IL-13 antibody construct as described in Example 114, wherein the anti-IL-13 scFv contains the amino acid sequence of SEQ ID NO:108.

實施例116.  如實施例113之分離的抗IL-13抗體構築體，其中該抗IL-13抗體部分為Fab (「抗IL-13 Fab」)。Example 116. An isolated anti-IL-13 antibody construct as described in Example 113, wherein the anti-IL-13 antibody portion is Fab ("anti-IL-13 Fab").

實施例117.  如實施例116之分離的抗IL-13抗體構築體，其中該抗IL-13 Fab包含含有SEQ ID NO:124之胺基酸序列的第一多肽及含有SEQ ID NO:102或197之胺基酸序列的第二多肽。Example 117. An isolated anti-IL-13 antibody construct as described in Example 116, wherein the anti-IL-13 Fab comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197.

實施例118.  如實施例113之分離的抗IL-13抗體構築體，其中該抗IL-13抗體部分為全長抗體(「抗IL-13全長抗體」)。Example 118. An isolated anti-IL-13 antibody construct as in Example 113, wherein the anti-IL-13 antibody portion is a full-length antibody ("anti-IL-13 full-length antibody").

實施例119.  如實施例118之分離的抗IL-13抗體構築體，其中該抗IL-13全長抗體包含： i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； ii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； iii)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈； iv)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； v)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； vi)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈； vii)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； viii)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； ix)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或 x)含有SEQ ID NO:130之胺基酸序列的第一重鏈、含有SEQ ID NO:131之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。Example 119. An isolated anti-IL-13 antibody construct according to Example 118, wherein the full-length anti-IL-13 antibody comprises:i) two heavy chains each containing the amino acid sequence of SEQ ID NO: 125 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197;ii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 101 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197;iii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 123 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206;iv) two heavy chains each containing the amino acid sequence of SEQ ID NO: 209 and two light chains each containing the amino acid sequence of SEQ ID NO: NO:122 or 207;v) two heavy chains each containing the amino acid sequence of SEQ ID NO:210 and two light chains each containing the amino acid sequence of SEQ ID NO:122 or 207;vi) two heavy chains each containing the amino acid sequence of SEQ ID NO:211 and two light chains each containing the amino acid sequence of SEQ ID NO:121 or 206;vii) two heavy chains each containing the amino acid sequence of SEQ ID NO:212 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208;viii) two heavy chains each containing the amino acid sequence of SEQ ID NO:213 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208;ix) two heavy chains each containing the amino acid sequence of SEQ ID NO:214 and two light chains each containing the amino acid sequence of SEQ ID NO:120 or 208; A heavy chain containing the amino acid sequence of SEQ ID NO: 225 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197; orx) a first heavy chain containing the amino acid sequence of SEQ ID NO: 130, a second heavy chain containing the amino acid sequence of SEQ ID NO: 131, and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197.

實施例120.  如請求項109至119中任一項之分離的抗IL-13抗體構築體，其中該分離的抗IL-13抗體構築體係單特異性。Embodiment 120. An isolated anti-IL-13 antibody construct as claimed in any one of claims 109 to 119, wherein the isolated anti-IL-13 antibody construct is monospecific.

實施例121.  如請求項109至119中任一項之分離的抗IL-13抗體構築體，其中該分離的抗IL-13抗體構築體係多特異性。Embodiment 121. An isolated anti-IL-13 antibody construct as claimed in any one of claims 109 to 119, wherein the isolated anti-IL-13 antibody construct is multispecific.

實施例122.  如實施例121之分離的抗IL-13抗體構築體，其中該分離的抗IL-13抗體構築體進一步包含特異性地結合至第二抗原的第二抗體部分。Example 122. The isolated anti-IL-13 antibody construct of Example 121, wherein the isolated anti-IL-13 antibody construct further comprises a second antibody portion that specifically binds to a second antigen.

實施例123.  如實施例122之分離的抗IL-13抗體構築體，其中該抗IL-13抗體部分及該第二抗體部分係經由連接子彼此融合。Example 123. The isolated anti-IL-13 antibody construct of Example 122, wherein the anti-IL-13 antibody portion and the second antibody portion are fused to each other via a linker.

實施例124.  如實施例123之分離的抗IL-13抗體構築體，其中該連接子含有GG及SEQ ID NO:98-99之任一者的胺基酸序列。Example 124. An isolated anti-IL-13 antibody construct as described in Example 123, wherein the linker contains GG and an amino acid sequence of any one of SEQ ID NOs: 98-99.

實施例125.  一種醫藥組成物，其包含如實施例109至124中任一者之分離的抗IL-13抗體構築體及醫藥上可接受之載體。Example 125. A pharmaceutical composition comprising the isolated anti-IL-13 antibody construct of any one of Examples 109 to 124 and a pharmaceutically acceptable carrier.

實施例126.  一種分離的核酸，其編碼如實施例109至124中任一者之分離的抗IL-13抗體構築體。Example 126. An isolated nucleic acid encoding an isolated anti-IL-13 antibody construct of any one of Examples 109 to 124.

實施例127.  一種載體，其包含如實施例126之分離的核酸。Example 127. A vector comprising the isolated nucleic acid of Example 126.

實施例128.  一種宿主細胞，其包含如實施例126之分離的核酸或如實施例127之載體。Example 128. A host cell comprising the isolated nucleic acid of Example 126 or the vector of Example 127.

實施例129.  一種治療個體之發炎性疾病的方法，其包含向該個體投予有效量之如實施例109至124中任一者之分離的抗IL-13抗體構築體或如實施例125之醫藥組成物。Example 129. A method for treating an inflammatory disease in a subject, comprising administering to the subject an effective amount of an isolated anti-IL-13 antibody construct of any one of Examples 109 to 124 or a pharmaceutical composition of Example 125.

實施例130.  如實施例129之方法，其中該發炎性疾病為氣喘、異位性皮膚炎或COPD。Example 130. The method of Example 129, wherein the inflammatory disease is asthma, atopic dermatitis, or COPD.

實施例131.  如實施例129或130之方法，其中該個體為人類。Embodiment 131. The method of embodiment 129 or 130, wherein the individual is a human.

實施例132.  一種產生分離的抗IL-13抗體構築體之方法，其包含： i)在適合表現該抗IL-13抗體構築體的條件下培養含有如實施例126之分離的核酸或如實施例127之載體的宿主細胞或如實施例128之宿主細胞；及 ii)獲得該表現的抗IL-13抗體構築體。 實例Example 132. A method for producing an isolated anti-IL-13 antibody construct, comprising:i) culturing a host cell containing the isolated nucleic acid of Example 126 or the vector of Example 127, or the host cell of Example 128, under conditions suitable for expression of the anti-IL-13 antibody construct; andii) obtaining the expressed anti-IL-13 antibody construct.Example

以下實施例僅用於示例本發明，因此不應被視為以任何方式限制本發明。以下實例及詳細描述僅用於說明目的而非限制。針對未描述實驗方法細節的實施例，此類方法係根據常規條件進行，諸如在Sambrook等人之Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press,1989)中描述的條件，或按照製造商建議的條件。實例1.示例性抗TSLP及抗IL-13抗體之產生及表徵。抗TSLP抗體The following examples are intended to illustrate the present invention only and should not be construed as limiting the present invention in any way. The following examples and detailed descriptions are for illustrative purposes only and are not intended to be limiting. For examples where the details of the experimental methods are not described, such methods were performed according to conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or in accordance with the conditions recommended by the manufacturer.Example1. Generation and characterization ofexemplary anti-TSLPand anti-IL-13antibodies.Anti-TSLPantibodies

TSLP藉由與TSLPR鏈結合進行傳訊，隨後招募IL-7Rα鏈並形成功能性TSLP受體複合體。為了破壞此動態並抑制TSLP傳訊，使用人類TSLP及食蟹獼猴(以下稱為「cyno」) TSLP作為免疫原來產生抗TSLP抗體。在確定個別動物具有適合的抗體滴度後，從免疫接種小鼠的脾臟及淋巴結中獲得淋巴細胞的單細胞懸浮液。藉由標準方法將淋巴球與鼠科骨髓瘤細胞融合以產生融合瘤。藉由抗TSLP阻斷抗體的報導細胞試驗進一步篩選出與人類及食蟹獼猴TSLP結合的融合瘤。TSLP signals by binding to the TSLPR chain, which subsequently recruits the IL-7Rα chain to form a functional TSLP receptor complex. To disrupt this dynamic and inhibit TSLP signaling, anti-TSLP antibodies were raised using human TSLP and cynomolgus macaque (hereafter referred to as "cyno") TSLP as immunogens. After confirming that the individual animals had suitable antibody titers, single-cell suspensions of lymphocytes were obtained from the spleens and lymph nodes of the immunized mice. The lymphocytes were fused with murine myeloma cells using standard methods to generate fusion tumors. Fusion tumors that bound to human and cynomolgus macaque TSLP were further screened using reporter cell assays with anti-TSLP blocking antibodies.

TSLP報導細胞試驗。TSLP報導細胞試驗確定了抗TSLP抗體在共同表現TSLPR/IL-7Rα的STAT5-螢光素酶HEK-293報導細胞中對TSLP的阻斷程度，其中STAT5傳訊增加會誘導螢光素酶產量增加。已知TSLP與TSLP受體複合體的結合可活化STAT5傳訊路徑。因此，藉由測量螢光素酶活性來監控STAT5傳訊路徑的活化，可確定TSLP阻斷程度。將具有TSLPR/IL-7Rα共同表現的STAT5-螢光素酶HEK-293報導細胞與10 ng/mL人類TSLP及本文所述之連續稀釋的抗TSLP抗體同時在37°C下培養5小時。在共同培養5小時之後，測量生物發光訊號。圖1顯示了使用GraphPad Prism分析的報導細胞的螢光素酶抑制百分比(IC90)結果，其證實所有示例性抗TSLP抗體殖株皆能有效阻斷游離TSLP與TSLP受體複合體的結合。TSLPreporter cell assay . The TSLP reporter cell assay determines the extent of TSLP blockade by anti-TSLP antibodies in STAT5-luciferase HEK-293 reporter cells co-expressing the TSLPR/IL-7Rα, where increased STAT5 signaling induces increased luciferase production. Binding of TSLP to the TSLP receptor complex is known to activate the STAT5 signaling pathway. Therefore, monitoring STAT5 signaling activation by measuring luciferase activity can determine the extent of TSLP blockade. STAT5-luciferase HEK-293 reporter cells co-expressing the TSLPR/IL-7Rα were incubated with 10 ng/mL human TSLP and serially diluted anti-TSLP antibodies as described herein for 5 hours at 37°C. After 5 hours of co-culture, bioluminescence signals were measured.Figure1 shows the percentage inhibition (IC90 ) of luciferase in reporter cells analyzed using GraphPad Prism, demonstrating that all exemplary anti-TSLP antibody clones were able to effectively block the binding of free TSLP to the TSLP receptor complex.

基於所選的小鼠抗TSLP單株抗體(mAb)建構出含有人類恆定域的重組嵌合抗TSLP mAb。A recombinant chimeric anti-TSLP mAb containing a human constant domain was constructed based on the selected mouse anti-TSLP monoclonal antibody (mAb).

使用Biacore來進行靶向TSLP之示例性融合瘤抗體之結合親和力的表面電漿共振(SPR)測量。簡言之，利用抗Fc抗體晶片來捕獲純化的抗體(亦即，針對具有人類Fc的重組抗體採用抗人類Fc晶片，且針對融合瘤或重組小鼠抗體採用抗小鼠Fc晶片)。抗原以不同的濃度(以1：3進行連續稀釋，從48 nM至0.2 nM)流過晶片。捕獲的抗原在900至1200秒內從晶片上分離。將感測圖擬合至1：1結合模型。結果總結於下表1至3。Surface plasmon resonance (SPR) measurements of the binding affinity of exemplary fusion tumor antibodies targeting TSLP were performed using Biacore. Briefly, purified antibodies were captured using an anti-Fc antibody chip (i.e., an anti-human Fc chip was used for recombinant antibodies with a human Fc, and an anti-mouse Fc chip was used for fusion tumor or recombinant mouse antibodies). Antigen was flowed over the chip at varying concentrations (serial dilutions of 1:3, from 48 nM to 0.2 nM). The captured antigen was detached from the chip within 900 to 1200 seconds. The sensorgrams were fitted to a 1:1 binding model. The results are summarized inTables1to3 below.

從表1至3可看出，所有測試的示例性小鼠抗TSLP mAb與人類及食蟹獼猴TSLP皆表現出非常強的結合(對人類TSLP的結合更佳)。此外，嵌合化不會顯著影響抗TSLP抗體的結合親和力(例如，與51A4及Ch51A4相比)。表1. 來自小鼠融合瘤之純化抗體的抗TSLP結合親和力結果。殖株名稱人類ka (1/Ms)人類kd (1/s)人類KD (M)食蟹獼猴ka (1/Ms)-1食蟹獼猴kd (1/s)-1食蟹獼猴KD (M)51B21.07E+072.05E-051.92E-128.44E+061.03E-031.22E-1051A41.19E+076.34E-055.31E-129.37E+061.54E-031.65E-1051A11.19E+079.98E-058.38E-129.48E+062.39E-032.52E-1051A61.48E+071.52E-041.02E-111.10E+072.21E-032.01E-1051A31.26E+071.70E-041.35E-111.05E+072.11E-032.02E-10表2. 純化的重組嵌合抗體的人類TSLP 結合親和力結果。名稱ka (1/Ms)kd (1/s)KD (M)Ch54A2 mAb8.63E+063.50E-054.05E-12Ch54C4 mAb4.89E+062.61E-055.35E-12Ch51A4 mAb4.50E+065.78E-041.28E-10表3. 純化的重組嵌合抗體的食蟹獼猴TSLP結合親和力結果。捕獲1溶液ka (1/Ms)kd (1/s)KD (M)Ch51A4 mAb1.13E+071.73E-031.53E-10Ch54A2 mAb6.32E+061.87E-022.97E-09Ch54C4 mAb4.62E+053.29E-037.13E-09As can be seen fromTables1to3 , all tested exemplary mouse anti-TSLP mAbs exhibited very strong binding to both human and cynomolgus macaque TSLP (with better binding to human TSLP). Furthermore, chimerization did not significantly affect the binding affinity of the anti-TSLP antibodies (e.g., compared to 51A4 and Ch51A4).Table1. Anti-TSLP binding affinity results of purified antibodies from mouse hybridomas.Strain nameHumanka (1/Ms)Humankd (1/s)HumanKD (M)Crab-eating macaqueka (1/Ms)-1Crab-eating macaquekd (1/s)-1Crab-eating macaqueKD (M) 51B2 1.07E+07 2.05E-05 1.92E-12 8.44E+06 1.03E-03 1.22E-10 51A4 1.19E+07 6.34E-05 5.31E-12 9.37E+06 1.54E-03 1.65E-10 51A1 1.19E+07 9.98E-05 8.38E-12 9.48E+06 2.39E-03 2.52E-10 51A6 1.48E+07 1.52E-04 1.02E-11 1.10E+07 2.21E-03 2.01E-10 51A3 1.26E+07 1.70E-04 1.35E-11 1.05E+07 2.11E-03 2.02E-10Table2. Human TSLP binding affinity results of purified recombinant chimeric antibodies.Nameka (1/Ms)kd (1/s)KD (M) Ch54A2 mAb 8.63E+06 3.50E-05 4.05E-12 Ch54C4 mAb 4.89E+06 2.61E-05 5.35E-12 Ch51A4 mAb 4.50E+06 5.78E-04 1.28E-10Table3. Cynomolgus macaque TSLP binding affinity results of purified recombinant chimeric antibodies.Capture1solutionka (1/Ms)kd (1/s)KD (M) Ch51A4 mAb 1.13E+07 1.73E-03 1.53E-10 Ch54A2 mAb 6.32E+06 1.87E-02 2.97E-09 Ch54C4 mAb 4.62E+05 3.29E-03 7.13E-09

與參考抗TSLP抗體相比，確定51B2結合表位及效力：將不同濃度的51B2及US-FDA批准的抗TSLP參考抗體(「參考抗TSLP Ab #5」)與生物素化的TSLP預先培養2小時，之後添加至預先塗覆有參考抗TSLP Ab #5的ELISA盤上，以評估參考抗TSLP Ab #5與51B2之間的交叉競爭。使用惰性陰性對照抗體(Ab)作為非競爭性Ab。在ELISA盤進行短暫培養及清洗之後，藉由HRP-鏈黴親和素來檢測捕獲在盤上的生物素-TSLP。圖2的結果顯示了惰性陰性對照Ab不與TSLP結合，且參考抗TSLP Ab #5完全與自身競爭TSLP結合。圖2證實了51B2不完全與參考抗TSLP Ab #5競爭以防止其與TSLP結合。此等結果顯示，51B2具有不同於參考抗TSLP Ab #5的獨特結合表位。Determining51B2'sbinding epitope and potencycomparedto a reference anti-TSLP antibody: 51B2 and a US-FDA-approved reference anti-TSLP antibody ("Reference Anti-TSLP Ab #5") were pre-incubated with biotinylated TSLP at varying concentrations for 2 hours before being added to an ELISA plate pre-coated with Reference Anti-TSLP Ab #5 to assess cross-competition between Reference Anti-TSLP Ab #5 and 51B2. An inert negative control antibody (Ab) served as a non-competing Ab. After a brief incubation and washing of the ELISA plate, captured biotin-TSLP was detected using HRP-streptavidin. The results inFigure2 show that the inert negative control Ab does not bind to TSLP, and that the reference anti-TSLP Ab #5 completely competes with itself for TSLP binding.Figure2 confirms that 51B2 does not completely compete with the reference anti-TSLP Ab #5 to prevent its binding to TSLP. These results indicate that 51B2 has a unique binding epitope that is different from the reference anti-TSLP Ab #5.

在人類PBMC及分離的CD1c+樹突狀細胞中測試51B2抑制初代人類細胞TSLP傳訊的效力。簡言之，將人類PBMC及分離的CD1c+樹突細胞接種於96孔盤中，並在多種濃度的51B2或參考抗TSLP Ab #5存在下分別與50 ng/ml或5 ng/ml TSLP一起培養。在48小時(PBMC)或24小時(樹突狀細胞)後收集細胞培養上清液，並按製造商的方案使用市售ELISA套組來分析TSLP誘導的CCL17分泌。TSLP傳訊增強會誘導CCL17分泌增加，因此，當以TSLP細胞激素處理表現TSLP受體的促發炎性細胞時，CCL17分泌的減少或抑制可作為抑制TSLP與TSLPR複合體結合的讀數。使用GraphPad Prism分析及呈現數據。如圖3A(人類PBMC)及圖3B(CD1c+ 樹突細胞)所示，在人類PBMC及分離的CD1c+樹突細胞中，與參考抗TSLP Ab #5相比，51B2在每個測試的抗體濃度下皆表現出顯著抑制CCL17。因此，此等結果證實，51B2在阻斷促發炎性免疫細胞中的TSLP傳訊上比US-FDA批准的參考抗TSLP Ab #5更有效。The potency of 51B2 to inhibit TSLP signaling in primary human cells was tested in human PBMCs and isolated CD1c+ dendritic cells. Briefly, human PBMCs and isolated CD1c+ dendritic cells were seeded in 96-well plates and incubated with 50 ng/ml or 5 ng/ml TSLP in the presence of various concentrations of 51B2 or reference anti-TSLP Ab #5. Cell culture supernatants were collected after 48 hours (PBMCs) or 24 hours (dendritic cells) and analyzed for TSLP-induced CCL17 secretion using a commercially available ELISA kit according to the manufacturer's protocol. Enhanced TSLP signaling induces increased CCL17 secretion. Therefore, when pro-inflammatory cells expressing the TSLP receptor are treated with the TSLP cytokine, decreased or inhibited CCL17 secretion serves as a readout for inhibition of TSLP binding to the TSLPR complex. Data were analyzed and presented using GraphPad Prism. As shownin Figure3A (human PBMCs) andFigure3B (CD1c+ dendritic cells), 51B2 exhibited significant inhibition of CCL17 at every antibody concentration tested, compared to the reference anti-TSLP Ab #5, in both human PBMCs and isolated CD1c+ dendritic cells. Therefore, these results demonstrate that 51B2 is more potent than the US-FDA-approved reference anti-TSLP Ab #5 in blocking TSLP signaling in pro-inflammatory immune cells.

51B2的人源化及人源化51B2變體：藉由將重鏈(HC)及輕鏈(LC)的CDR移植至人類HC及LC框架序列上使親代殖株51B2人源化。簡言之，將嵌合51B2的HC及LC CDR移植至最接近的HC及LC人類種系上。利用目視檢查序列來識別就CDR結合及結構穩定性而言重要的框架殘基。將此等框架殘基進行單獨或組合突變。參見針對VL序列的SEQ ID NO:51-54及針對VH序列的SEQ ID NO:55-64。Humanization of51B2and Humanized51B2Variants : The parental strain 51B2 was humanized by grafting the heavy chain (HC) and light chain (LC) CDRs onto human HC and LC framework sequences. Briefly, the HC and LC CDRs of the chimeric 51B2 were grafted onto the closest human germline HC and LC. Visual inspection of the sequence was used to identify framework residues important for CDR binding and structural stability. These framework residues were mutated individually or in combination. See SEQ ID NOs: 51-54 for VL sequences and SEQ ID NOs: 55-64 for VH sequences.

使用Biacore測試人源化51B2抗體的結合親和力，並根據活性(類似於上述之TSLP報導試驗)及表現品質/純化行為來進行排名。如表4所示，除了L1H2及L1H9變體以外，幾乎所有人源化51B2變體皆保留了與人類TSLP類似甚至表現出更強的結合親和力。基於Biacore結合結果(使用抗人類Fc捕獲晶片；參見表4至5)及活性試驗(參見圖4及圖5A)，將人源化51B2抗體、hz51B2-L3H9 (亦即，具有VL-3 (SEQ ID NO: 53)及VH-9 (SEQ ID NO: 63)配對)選為最終的抗TSLP先導殖株。表4. 人源化51B2變體與人類TSLP的結合親和力。LCHCka (1/Ms)kd (1/s)KD (M)Ch51B2Ch51B24.37E+072.41E-045.52E-12L1H11.20E+073.33E-042.78E-11L1H27.05E+062.64E-013.75E-08L1H91.05E+072.98E-012.85E-08L1H105.11E+081.78E-033.49E-12L2H12.76E+111.21E+004.38E-12L2H24.35E+073.31E-037.62E-11L2H95.27E+076.51E-041.24E-11L3H35.81E+071.40E-022.41E-10L3H48.68E+071.20E-031.39E-11L3H81.80E+081.04E-035.78E-12L3H98.67E+071.07E-041.24E-12L3H105.13E+083.36E-046.54E-13L4H18.36E+062.38E-042.85E-11L4H77.81E+062.78E-043.55E-11L4H85.41E+061.68E-063.11E-13L4H95.41E+074.23E-047.82E-12L4H104.27E+071.88E-044.41E-12表5. hz51B2-L3H9抗TSLP抗體與人類及食蟹獼猴TSLP的結合親和力。星號(*)標記達到Biacore解析極限的值。抗體抗原ka (1/Ms)kd (1/s)KD (M)hz51B2-L3H9huTSLP8.67E+071.07E-041.24E-12*hz51B2-L3H9食蟹獼猴TSLP1.20E+08*1.04E-028.66E-11The binding affinity of humanized 51B2 antibodies was tested using Biacore and ranked based on activity (similar to the TSLP reporter assay described above) and performance quality/purification behavior. As shown inTable4 , with the exception of the L1H2 and L1H9 variants, nearly all humanized 51B2 variants retained similar or even stronger binding affinity to human TSLP. Based on Biacore binding results (using an anti-human Fc capture chip;seeTables4and5 ) and activity assays (seeFigures4 and5A ), the humanized 51B2 antibody, hz51B2-L3H9 (i.e., with a VL-3 (SEQ ID NO: 53) and VH-9 (SEQ ID NO: 63) pairing), was selected as the final anti-TSLP lead clone.Table4. Binding affinity of humanized 51B2 variants to human TSLP.LCHCka (1/Ms)kd (1/s)KD (M)Ch51B2Ch51B2 4.37E+07 2.41E-04 5.52E-12 L1 H1 1.20E+07 3.33E-04 2.78E-11 L1 H2 7.05E+06 2.64E-01 3.75E-08 L1 H9 1.05E+07 2.98E-01 2.85E-08 L1 H10 5.11E+08 1.78E-03 3.49E-12 L2 H1 2.76E+11 1.21E+00 4.38E-12L2H2 4.35E+07 3.31E-03 7.62E-11L2H9 5.27E+07 6.51E-04 1.24E-11 L3 H3 5.81E+07 1.40E-02 2.41E-10 L3 H4 8.68E+07 1.20E-03 1.39E-11 L3 H8 1.80E+08 1.04E-03 5.78E-12L3H9 8.67E+07 1.07E-04 1.24E-12 L3 H10 5.13E+08 3.36E-04 6.54E-13 L4 H1 8.36E+06 2.38E-04 2.85E-11 L4 H7 7.81E+06 2.78E-04 3.55E-11 L4 H8 5.41E+06 1.68E-06 3.11E-13L4H9 5.41E+07 4.23E-04 7.82E-12 L4 H10 4.27E+07 1.88E-04 4.41E-12Table5. Binding affinity of hz51B2-L3H9 anti-TSLP antibody to human and cynomolgus macaque TSLP. Asterisks (*) indicate values that reached the Biacore resolution limit.antibodyantigenka (1/Ms)kd (1/s)KD (M) hz51B2-L3H9 huTSLP 8.67E+07 1.07E-04 1.24E-12* hz51B2-L3H9 TSLP in cynomolgus macaques 1.20E+08* 1.04E-02 8.66E-11

表5總結的結果顯示人源化hz51B2 L3H9抗體與人類TSLP及食蟹獼猴TSLP皆有強的結合，其有利於將食蟹獼猴毒性及功效研究的結果推斷至人類臨床研究中，以評估抗TSLP抗體或包含其之構築體(諸如本文所述之多特異性抗IL-13×TSLP抗體構築體之任一者)。The results summarized inTable5 show that the humanized hz51B2 L3H9 antibody strongly binds to both human TSLP and cynomolgus macaque TSLP. This facilitates the extrapolation of the results of cynomolgus macaque toxicity and efficacy studies to human clinical studies for the evaluation of anti-TSLP antibodies or constructs comprising them (such as any of the multispecific anti-IL-13 × TSLP antibody constructs described herein).

藉由使用BiacoreT200儀器測量平衡結合反應混合物中之游離TSLP濃度來表徵hz51B2 L3H9與人類TSLP的結合。簡言之，在室溫下，在含有0.1 mg/ml BSA及0.005%聚山梨醇20的PBS溶液中，將1 nM人類TSLP與3×連續稀釋的hz51B2 L3H9溶液(範圍從96 nM至44 pM的Fab臂)培養過夜。藉由將結合溶液注射至胺固定的hz51B2 L3H9表面上方來測量各個結合反應中未結合的TSLP。使用GraphPad將各個反應混合物中的游離TSLP濃度百分比擬合至1:1平衡Ab-Ag結合模型，並從曲線擬合推導出KD(圖5B)。Binding of hz51B2 L3H9 to human TSLP was characterized by measuring the concentration of free TSLP in the equilibrium binding reaction mixture using a Biacore T200 instrument. Briefly, 1 nM human TSLP was incubated with 3x serially diluted hz51B2 L3H9 solutions (ranging from 96 nM to 44 pM Fab arms) in PBS containing 0.1 mg/ml BSA and 0.005% polysorbate 20 overnight at room temperature. Unbound TSLP was measured in each binding reaction by injecting the binding solution over the amine-immobilized hz51B2 L3H9 surface. The percentagefree TSLP concentration in each reaction mixture was fit to a 1:1 equilibrium Ab-Ag binding model using GraphPad, and the KD was deduced from the curve fit (FIG5B ).

總之，先導51B2抗體(例如，hz51B2 Ab，諸如hz51B2 L3H9)為人類TSLP的緊密結合抗體，其中KD為＜ 10 pM。不受理論的束縛，此種性質使得在PBMC及樹突細胞試驗中能夠完全中和TSLP誘導的CCL17 (參見圖3A-3B)，而US-FDA批准的參考Ab. #1則不完全抑制TSLP誘導的CCL1。抗IL-13抗體In summary, the lead 51B2 antibodies (e.g., hz51B2 Ab, such as hz51B2 L3H9) are tight binding antibodies to human TSLP with aKD of < 10 pM. Without theoretical constraints, this property enables complete neutralization of TSLP-induced CCL17 in PBMC and dendritic cell assays (seeFigures3A-3B ), whereas the US-FDA-approved reference Ab. #1 does not completely inhibit TSLP-induced CCL1.Anti-IL-13Antibodies

為了參與IL-13傳訊，IL-13細胞激素首先與IL-13Rα1結合，隨後招募IL-4Rα以形成三元複合體。以下所述之示例性IL-13抗體藉由結合至IL-13並阻斷IL-4Rα的招募來抑制IL-13傳訊，從而阻斷三元複合體的形成。當抗IL-13抗體同時阻斷IL-13與IL-13Rα1及IL-13Rα2的結合時可稱為「第I類」，其中IL-13Rα2為IL-13誘餌受體並可活化單獨的細胞內傳訊級聯。當抗IL-13抗體阻斷IL-13與IL-4Rα的結合時可稱為「第II類」，例如可阻斷三元複合體的形成。阻斷IL-13與其誘餌受體IL-13Rα2的結合可增加血清IL-13水平，其可導致在臨床環境下療效及給藥的不理想。經由抑制IL-13/IL-13Rα1/IL-4Rα三元複合體的形成而非IL-13Rα2來阻斷IL-13活性可能表現出更好的臨床結果。舉例而言，參考抗IL-13抗體#2為第II類抗IL-13 Ab，其表現出比參考抗IL-13抗體#1 (第I類抗IL-13 Ab)更強的結合親和力及更高的IL-13抑制效力。參考抗IL-13抗體#1及參考抗IL-13抗體#2為US-FDA或EMA批准的抗IL-13抗體。To participate in IL-13 signaling, the IL-13 cytokine first binds to IL-13Rα1, which then recruits IL-4Rα to form a ternary complex. The exemplary IL-13 antibodies described below inhibit IL-13 signaling by binding to IL-13 and blocking the recruitment of IL-4Rα, thereby preventing the formation of the ternary complex. Anti-IL-13 antibodies that simultaneously block the binding of IL-13 to IL-13Rα1 and IL-13Rα2 are referred to as "Type I," where IL-13Rα2 is an IL-13 decoy receptor and can activate separate intracellular signaling cascades. Anti-IL-13 antibodies that block the binding of IL-13 to IL-4Rα are referred to as "Type II," for example, preventing the formation of the ternary complex. Blocking IL-13 binding to its decoy receptor, IL-13Rα2, increases serum IL-13 levels, which can lead to suboptimal therapeutic efficacy and dosing in clinical settings. Blocking IL-13 activity by inhibiting the formation of the IL-13/IL-13Rα1/IL-4Rα ternary complex, rather than IL-13Rα2, may demonstrate better clinical outcomes. For example, reference anti-IL-13 antibody #2 is a Class II anti-IL-13 Ab that exhibits stronger binding affinity and greater IL-13 inhibitory potency than reference anti-IL-13 antibody #1 (a Class I anti-IL-13 Ab). Reference anti-IL-13 antibody #1 and reference anti-IL-13 antibody #2 are US-FDA- or EMA-approved anti-IL-13 antibodies.

在參考抗IL-13抗體(「參考抗IL-13 Ab #0」)的CDR中產生突變，以便消除CDR缺陷並增強參考抗IL-13抗體針對食蟹獼猴IL-13的親和力。參考抗IL-13 Ab #0包含含有SEQ ID NO:101之胺基酸序列的重鏈及含有SEQ ID NO:116之胺基酸序列的輕鏈。參考抗IL-13 Ab #0含有SEQ ID NO:66之CDR-H1、SEQ ID NO:67之CDR-H2、SEQ ID NO:68之CDR-H3、SEQ ID NO:115之CDR-L1、SEQ ID NO:71之CDR-L2及SEQ ID NO:72之CDR-L3。參見US7585500，其內容通過引用整體併入本文。評估了抗體抗原界面，並識別出CDR-L1中之LC C34 (Kabat 編號)及CDR-H1中之HC M34 (Kabat編號)的位置以作為潛在的突變位點。突變僅導入所有6個CDR的CDR-L1中，或僅導入所有6個CDR的CDR-H1中。Mutations were generated in the CDRs of a reference anti-IL-13 antibody ("reference anti-IL-13 Ab #0") to eliminate CDR deficiencies and enhance the affinity of the reference anti-IL-13 antibody for cynomolgus macaque IL-13. Reference anti-IL-13 Ab #0 comprises a heavy chain comprising the amino acid sequence of SEQ ID NO: 101 and a light chain comprising the amino acid sequence of SEQ ID NO: 116. Reference anti-IL-13 Ab #0 comprises CDR-H1 of SEQ ID NO: 66, CDR-H2 of SEQ ID NO: 67, CDR-H3 of SEQ ID NO: 68, CDR-L1 of SEQ ID NO: 115, CDR-L2 of SEQ ID NO: 71, and CDR-L3 of SEQ ID NO: 72. See US7585500, the contents of which are incorporated herein by reference in their entirety. The antibody-antigen interface was evaluated, and the positions of LC C34 (Kabat numbering) in CDR-L1 and HC M34 (Kabat numbering) in CDR-H1 were identified as potential mutation sites. Mutations were introduced only in CDR-L1 or only in CDR-H1.

表6總結了位於位置C34處的輕鏈突變在瞬時表現滴度後的初步表現評估。未顯示CDR-H1變體的表達滴度數據。選擇了增強表現滴度的CDR-L1變體(C34Y、C34M、C34F及A33S/C34F)及CDR-H1變體，並評估了與參考抗IL-13 Ab #0相比針對食蟹獼猴TSLP結合的改善。使用Biacore T200分析參考抗IL-13 Ab #0、CDR-L1變體及CDR-H1變體的結合親和力。參見表7至8的結果。表6. 抗IL-13抗體CDR-L1變體的表現。CDR-L1SEQ ID NO:來自30 ml的重量(mg)​野生型SGDKLGDKYAC1152.83​突變SGDKLGDKYAY(「C34Y”；73P1)704.19​SGDKLGDKYAW(C34W)1464.06​SGDKLGDKYAL(C34L)1831.45SGDKLGDKYAI(C34I)1841.27SGDKLGDKYAM(C34M)1483.69​SGDKLGDKYAA(C34A)1854.02​SGDKLGDKYAS(C34S)1863.91​SGDKLGDKYAF(C34F)1194.45SGDKLGDKYAH(C34H)1873.02SGDKLGDKYSF(A33S/C34F)1822.63表7. 抗IL-13抗體CDR-L1變體與食蟹獼猴IL-13的結合親和力。CDR-L1SEQ ID NO:ka (1/Ms)kd (1/s)KD (M)SGDKLGDKYAC1157.36E+061.14E-021.55E-09SGDKLGDKYAY(「C34Y”；73P1)707.14E+061.81E-032.53E-10SGDKLGDKYAM(C34M)1486.11E+066.52E-031.07E-09SGDKLGDKYAF(C34F)1199.88E+063.22E-033.26E-10SGDKLGDKYSF(A33S/C34F)1827.86E+062.50E-033.26E-10表8. 抗IL-13抗體CDR-H1變體與食蟹獼猴IL-13的結合親和力。CDR-H1SEQ ID NO:ka (1/Ms)kd (1/s)KD (M)SYAMS662.72E+093.93E+001.45E-09SYAGS  (M34G)1506.08E+063.56E-045.86E-11SYAFS  (M34F)1525.92E+064.46E-037.54E-10SYASS  (M34S)1544.52E+061.73E-033.83E-10SYAYS  (M34Y)1564.48E+062.20E-024.92E-09Table6 summarizes the preliminary performance evaluation of light chain mutations at position C34 following transient titer analysis. Titer data for CDR-H1 variants are not shown. CDR-L1 variants (C34Y, C34M, C34F, and A33S/C34F) and CDR-H1 variants that enhanced titer were selected and evaluated for improved binding to cynomolgus macaque TSLP compared to reference anti-IL-13 Ab #0. Binding affinities of reference anti-IL-13 Ab #0, CDR-L1 variants, and CDR-H1 variants were analyzed using Biacore T200.See the results inTables 7and8.Table6. Performance of anti-IL-13 antibody CDR-L1 variants. CDR-L1SEQ ID NO: Weight from 30 ml(mg) wild type SGDKLGDKYAC 1152.83 mutation SGDKLGDKYAY (“C34Y”; 73P1) 70 4.19 SGDKLGDKYAW (C34W) 146 4.06 SGDKLGDKYAL (C34L) 183 1.45 SGDKLGDKYAI (C34I) 184 1.27 SGDKLGDKYAM (C34M) 148 3.69 SGDKLGDKYAA (C34A) 185 4.02 SGDKLGDKYAS (C34S) 186 3.91 SGDKLGDKYAF (C34F) 119 4.45 SGDKLGDKYAH (C34H) 187 3.02 SGDKLGDKYSF (A33S/C34F) 182 2.63Table7. Binding affinity of anti-IL-13 antibody CDR-L1 variants to cynomolgus macaque IL-13.CDR-L1SEQ ID NO:ka (1/Ms)kd (1/s)KD (M) SGDKLGDKYAC 115 7.36E+06 1.14E-02 1.55E-09 SGDKLGDKYAY (“C34Y”; 73P1) 70 7.14E+06 1.81E-03 2.53E-10 SGDKLGDKYAM (C34M) 148 6.11E+06 6.52E-03 1.07E-09 SGDKLGDKYAF (C34F) 119 9.88E+06 3.22E-03 3.26E-10 SGDKLGDKYSF (A33S/C34F) 182 7.86E+06 2.50E-03 3.26E-10Table8. Binding affinity of anti-IL-13 antibody CDR-H1 variants to cynomolgus macaque IL-13.CDR-H1SEQ ID NO:ka (1/Ms)kd (1/s)KD (M) SYAMS 66 2.72E+09 3.93E+00 1.45E-09 SYAG S (M34G) 150 6.08E+06 3.56E-04 5.86E-11 SYAF S (M34F) 152 5.92E+06 4.46E-03 7.54E-10 SYAS S (M34S) 154 4.52E+06 1.73E-03 3.83E-10 SYAY S (M34Y) 156 4.48E+06 2.20E-02 4.92E-09

具有C34Y CDR-L1突變的抗IL-13變體與食蟹獼猴IL-13的結合增強。此殖株亦顯示出與人類IL-13的結合非常強，類似於參考抗IL-13 Ab #0 (表9)。此抗體在下方稱為殖株「73P1」並用於後續分析。73P1 mAb包含含有SEQ ID NO:101之胺基酸序列的重鏈及含有SEQ ID NO:102或197之胺基酸序列的輕鏈。表9. 73P1針對人類及食蟹獼猴IL-13的結合表徵。IL-13抗體ka (1/Ms)kd (1/s)KD (M)人類參考抗IL-13 Ab #03.15E+062.12E-046.74E-1173P12.99E+061.41E-044.71E-11食蟹獼猴參考抗IL-13 Ab #02.73E+093.93E+001.45E-0973P17.14E+061.83E-032.53E-10Anti-IL-13 variants with the C34Y CDR-L1 mutation showed enhanced binding to cynomolgus macaque IL-13. This strain also showed very strong binding to human IL-13, similar to reference anti-IL-13 Ab #0 (Table9 ). This antibody is referred to below as strain "73P1" and was used in subsequent analyses. The 73P1 mAb comprises a heavy chain containing the amino acid sequence of SEQ ID NO: 101 and a light chain containing the amino acid sequence of SEQ ID NO: 102 or 197.Table9. Binding characteristics of 73P1 against human and cynomolgus macaque IL-13.IL-13antibodyka (1/Ms)kd (1/s)KD (M)HumanReference anti-IL-13 Ab #0 3.15E+06 2.12E-04 6.74E-1173P1 2.99E+06 1.41E-04 4.71E-11Crab-eating macaqueReference anti-IL-13 Ab #0 2.73E+09 3.93E+00 1.45E-0973P1 7.14E+06 1.83E-03 2.53E-10

使用表現IL-13Rα1及IL-4Rα的HEK-Blue 293報導細胞來測試抗IL-13抗體殖株73P1及參考抗IL-13 Ab #0針對IL-13的中和作用。在此試驗中，當IL-13結合至由HEK-Blue 293報導細胞表現的IL-13Rα1及IL-4Rα時，細胞內IL-13傳訊會活化JAK/STAT6路徑，進而導致產生分泌的胚胎鹼性磷酸酯酶(SEAP)。簡言之，同時將HEK-Blue 293報導細胞與10 ng/ml人類IL-13 (圖6A)或10 ng/ml食蟹獼猴(「cyno」) IL-13 (圖6B)及連續稀釋的參考抗IL-13 Ab #0及73P1在37°C下培養過夜。在培養之後，使用QU抗Blue溶液及市售盤讀儀來測定SEAP含量。使用SEAP的抑制百分比來呈現數據(圖6A至6B)。如圖6A所示，抗IL-13抗體殖株73P1 (IC50=7.4 pM)顯示與參考抗IL-13 Ab #0 (IC50=7 pM)有類似的人類IL-13中和作用。令人驚訝的是，與圖6B的參考抗IL-13 Ab #0 (IC50=2.39 nM)相比，73P1 (IC50=0.37 nM)顯示顯著改善食蟹獼猴IL-13的中和作用，其可有助於將食蟹獼猴的毒性及功效研究結果推斷至人體臨床研究。The neutralization of IL-13 by the anti-IL-13 antibody clone 73P1 and the reference anti-IL-13 Ab #0 was tested using HEK-Blue 293 reporter cells expressing IL-13Rα1 and IL-4Rα. In this assay, when IL-13 binds to IL-13Rα1 and IL-4Rα expressed by HEK-Blue 293 reporter cells, intracellular IL-13 signaling activates the JAK/STAT6 pathway, leading to the production of secreted embryonic alkaline phosphatase (SEAP). Briefly, HEK-Blue 293 reporter cells were incubated overnight at 37°C with 10 ng/ml human IL-13 (Figure6A ) or 10 ng/ml cynomolgus macaque ("cyno") IL-13 (Figure6B ) and serially diluted reference anti-IL-13 Abs #0 and 73P1. Following incubation, SEAP levels were measured using QU Anti-Blue solution and a commercial plate reader. Data are presented as percent inhibition of SEAP(Figures6Aand6B ). As shown inFigure6A , anti-IL-13 Ab 73P1 (IC50 = 7.4 pM) exhibited similar neutralization of human IL-13 as reference anti-IL-13 Ab #0 (IC50 = 7 pM). Surprisingly, 73P1 (IC50 =0.37 nM) demonstrated significantly improved neutralization of cynomolgus macaque IL-13 compared to the reference anti-IL-13 Ab #0 (IC50 =2.39 nM) inFigure6B , which may facilitate the extrapolation of cynomolgus macaque toxicity and efficacy findings to human clinical studies.

使用上述之HEK-Blue 293報導細胞來測試抗IL-13抗體殖株73P1、第I類參考抗IL-13抗體(「參考Ab.」) #1及第II類參考抗IL-13 Ab. #2針對IL-13的中和作用。簡言之，利用濃度為1.6 nM的人類野生型IL-13 (圖7A)或濃度為0.8 nM的疾病相關IL-13變體R110Q (其表現出IL-13活性增加)(圖7B)來刺激HEK-293報導細胞。所計算出的IC50值表明，73P1比參考抗IL-13 Ab. #1或#2在抑制野生型及突變體 IL-13上更有效。此外，從圖7A至7B中可看出，73P1與第II類參考抗IL-13 Ab. #2相比具有類似(但更好)的IL-13中和活性，且與第I類參考抗IL-13 Ab. #1相比具有更好的IL-13中和活性，其顯示73P1可發揮第II類抗IL-13抗體的功用，亦即阻斷與IL-4Rα形成IL-13/IL-13Rα複合體。實例2.靶向TSLP及IL-13之示例性雙特異性抗體構築體的表徵。The anti-IL-13 antibody clone 73P1, class I reference anti-IL-13 antibody ("Reference Ab") #1, and class II reference anti-IL-13 Ab #2 were tested for their IL-13 neutralization activity using HEK-Blue 293 reporter cells described above. Briefly, HEK-293 reporter cells were stimulated with either 1.6 nM human wild-type IL-13 (Figure7A ) or 0.8 nM of the disease-relevant IL-13 variant R110Q (which exhibits increased IL-13 activity) (Figure7B ). CalculatedIC50 values demonstrated that 73P1 was more potent than either reference anti-IL-13 Ab #1 or #2 in inhibiting both wild-type and mutant IL-13. Furthermore, as can be seen fromFigures7Aand7B , 73P1 has similar (but superior) IL-13 neutralizing activity compared to Class II reference anti-IL-13 Ab #2, and superior IL-13 neutralizing activity compared to Class I reference anti-IL-13 Ab #1, demonstrating that 73P1 can function as a Class II anti-IL-13 antibody, namely, blocking the formation of an IL-13/IL-13Rα complex with IL-4Rα.Example2.Characterization of exemplary bispecific antibody constructstargetingTSLPandIL-13 .

利用scFv及Fab或全長抗體設計的組合來產生靶向TSLP及IL-13的示例性雙特異性抗體構築體，如圖8A至8D所示，使用hz51B2-L3H9作為抗TSLP Fab或全長抗體部分，或hz51B2-L4H9作為抗TSLP scFv部分，以及73P1作為抗IL-13抗體部分。圖8A描繪了PX128-A設計，其中雙特異性抗體構築體包含具有兩個各自附接至抗TSLP抗體重鏈之C端的抗IL-13 73P1 scFv的全長抗TSLP hz51B2 L3H9抗體。圖8B描繪了PX128-B設計，其中該雙特異性抗體包含N端至C端各自含有以下的兩個融合配對：抗TSLP hz51B2-L3H9 Fab、融合至抗TSLP Fab之CH1之C端的抗IL-13 73P1 scFv及融合至抗IL-13 scFv之C端的Fc域之次單元。PX128-B亦可被視為雙特異性抗體構築體，其包含具有兩個各自在全長抗TSLP抗體之鉸鏈區的次單元處插入抗IL-13 scFv的全長抗TSLP抗體。圖8C描繪了PX128-C設計，其中雙特異性抗體構築體包含具有兩個各自融合至抗IL-13抗體重鏈之C端的抗TSLP hz51B2-L4H9 scFv的全長抗IL-13 73P1抗體。圖8D描繪了PX128-D設計，其中該雙特異性抗體包含N端至C端各自含有以下的兩個融合配對：抗IL-13 73P1 Fab、融合至抗IL-13 Fab之CH1之C端的抗TSLP hz51B2-L4H9 scFv及融合至抗TSLP scFv之C端的Fc域之次單元。PX128-D亦可被視為雙特異性抗體構築體，其包含具有兩個各自在全長抗IL-13抗體之鉸鏈區的次單元處插入抗TSLP scFv的全長抗IL-13抗體。該構築體之任一者可進一步包括框架區、CH1、CL、鉸鏈及/或Fc域的突變，例如延長抗體半衰期的突變。Exemplary bispecific antibody constructs targeting TSLP and IL-13 were generated using a combination of scFv and Fab or full-length antibody designs, as shown inFigures8Ato8D , using hz51B2-L3H9 as the anti-TSLP Fab or full-length antibody portion, or hz51B2-L4H9 as the anti-TSLP scFv portion, and 73P1 as the anti-IL-13 antibody portion.Figure8A depicts the PX128-A design, in which the bispecific antibody construct comprises a full-length anti-TSLP hz51B2 L3H9 antibody with two anti-IL-13 73P1 scFvs, each attached to the C-terminus of the anti-TSLP antibody heavy chain.Figure8B depicts the design of PX128-B, in which the bispecific antibody comprises two fusion partners from the N-terminus to the C-terminus: an anti-TSLP hz51B2-L3H9 Fab, an anti-IL-13 73P1 scFv fused to the C-terminus of the CH1 of the anti-TSLP Fab, and a subunit with the Fc domain fused to the C-terminus of the anti-IL-13 scFv. PX128-B can also be considered a bispecific antibody construct, comprising a full-length anti-TSLP antibody with two subunits, each containing an anti-IL-13 scFv inserted into the hinge region of the full-length anti-TSLP antibody.Figure8C depicts the PX128-C design, in which the bispecific antibody construct comprises a full-length anti-IL-13 73P1 antibody with two anti-TSLP hz51B2-L4H9 scFvs, each fused to the C-terminus of the anti-IL-13 antibody heavy chain.Figure8D depicts the PX128-D design, in which the bispecific antibody comprises two fusion partners, each containing the following from the N-terminus to the C-terminus: an anti-IL-13 73P1 Fab, an anti-TSLP hz51B2-L4H9 scFv fused to the C-terminus of the CH1 of the anti-IL-13 Fab, and a subunit of the Fc domain fused to the C-terminus of the anti-TSLP scFv. PX128-D can also be considered a bispecific antibody construct comprising a full-length anti-IL-13 antibody with two subunits, each containing an anti-TSLP scFv, inserted into the hinge region of the full-length anti-IL-13 antibody. Any of these constructs may further include mutations in the framework, CH1, CL, hinge, and/or Fc domains, e.g., mutations that increase antibody half-life.

將編碼構築體PX128-A、PX128-B、PX128-C及PX128-D之多肽鏈的核酸轉染至HEK-293細胞中，以產生雙特異性抗體構築體多肽並分泌至培養基中。在轉染後7天收穫澄清的條件培養基以分離分泌的多肽。隨後，在蛋白A管柱上捕獲雙特異性分子以進行純化。藉由PBS清洗管柱來移除宿主衍生的雜質，並藉由酸性溶析從管柱中回收雙特異性分子。隨後，將蛋白A純化的雙特異性分子中和，並在陶瓷羥基磷灰石(CHT)或Capto MMC ImpRes的混合模式層析管柱上進一步純化。Nucleic acids encoding the polypeptide chains of constructs PX128-A, PX128-B, PX128-C, and PX128-D were transfected into HEK-293 cells, resulting in the production of bispecific antibody construct polypeptides, which were secreted into the culture medium. Seven days after transfection, the clarified conditioned medium was harvested to isolate the secreted polypeptides. The bispecific molecules were then captured on a Protein A column for purification. The column was washed with PBS to remove host-derived impurities, and the bispecific molecules were recovered from the column by acidic elution. Subsequently, the protein A-purified bispecific molecules were neutralized and further purified on ceramic hydroxyapatite (CHT) or Capto MMC ImpRes mixed-mode chromatography columns.

基於scFv的雙特異性抗體在報導細胞試驗中針對TSLP及IL-13的活性：如實例1所述，在共同表現TSLPR/IL-7Rα的HEK-293報導細胞中評估雙特異性抗體對人類TSLP的阻斷。簡言之，同時將TSLP HEK-293報導細胞與10 ng/mL人類TSLP及連續稀釋的上述各個PX128雙特異性抗體在37°C下培養5小時。在共同培養5小時之後，測量生物發光訊號。數據以細胞激素-螢光素酶活性的抑制百分比呈現於圖9A中，並證實所有示例性IL-13×TSLP雙特異性抗體形式皆能經由TSLPR/IL-7Rα受體複合體有效阻斷TSLP傳訊，其中PX128-A在較低濃度下顯示出最高的抑制活性(IC90=6.04 nM)。與其他雙特異性抗體相比，位於抗IL-13 Fab與Fc域之間的具有抗TSLP scFv的PX128-D顯示出相對較低的TSLP抑制活性(IC90=18.1 nM)，此可能係因位置效應掩蓋了抗TSLP scFv 與TSLP的結合。Activity ofscFv -basedbispecific antibodies againstTSLPandIL-13 in a reporter cell assay: As described in Example 1, bispecific antibody blockade of human TSLP was evaluated in HEK-293 reporter cells co-expressing TSLPR/IL-7Rα. Briefly, TSLP HEK-293 reporter cells were incubated with 10 ng/mL human TSLP and serially diluted versions of each of the PX128 bispecific antibodies listed above for 5 hours at 37°C. Bioluminescence signal was measured after 5 hours of co-incubation. The data, presented as percent inhibition of cytokine-luciferase activity inFigure9A , demonstrate that all exemplary IL-13×TSLP bispecific antibody formats effectively blocked TSLP signaling through the TSLPR/IL-7Rα receptor complex, with PX128-A exhibiting the highest inhibitory activity at lower concentrations (IC90 = 6.04 nM). Compared to the other bispecific antibodies, PX128-D, which harbors an anti-TSLP scFv positioned between the anti-IL-13 Fab and Fc domains, exhibited relatively low TSLP inhibitory activity (IC90 = 18.1 nM), likely due to positional effects that masked TSLP binding.

同樣地，如實例1所述，HEK-Blue 293報導細胞(其細胞表面上表現人類IL-13Rα1及IL-4Rα且在IL-13傳訊路徑控制下以SEAP穩定轉染)用於研究測試抗體的IL-13中和能力。簡言之，同時將IL-13 HEK-293報導細胞與10 ng/mL人類IL-13及連續稀釋的上述各個PX128雙特異性抗體在37°C下培養過夜。在共同培養之後，測量SEAP訊號。數據以SEAP活性的抑制百分比呈現於圖9B中，並證實所有示例性PX128 IL-13×TSLP雙特異性抗體形式皆能經由IL-13Rα1/IL-4Rα受體複合體有效阻斷IL-13傳訊。 實例3：示例性雙特異性IL-13×TSLP抗體構築體的小鼠藥物動力學。Similarly, as described in Example 1, HEK-Blue 293 reporter cells (which express human IL-13Rα1 and IL-4Rα on their cell surface and are stably transfected with SEAP under the control of the IL-13 signaling pathway) were used to investigate the IL-13 neutralization ability of the test antibodies. Briefly, IL-13 HEK-293 reporter cells were co-cultured with 10 ng/mL human IL-13 and serially diluted versions of each of the aforementioned PX128 bispecific antibodies overnight at 37°C. Following co-culture, SEAP signaling was measured. The data are presented inFigure9B as percent inhibition of SEAP activity and demonstrate that all exemplary PX128 IL-13 × TSLP bispecific antibody formats can effectively block IL-13 signaling through the IL-13Rα1/IL-4Rα receptor complex. Example 3:Mouse Pharmacokinetics of ExemplaryBispecificIL-13 × TSLP Antibody Constructs.

測試上述PX128 IL-13×TSLP雙特異性抗體，以評估平均藥物動力學特徵並驗證各個構築體的體內穩定性。簡言之，C57BL/6小鼠以1 mg/kg的單一劑量雙特異性抗體進行靜脈內注射，並根據投予的雙特異性抗體進行分組。在5分鐘、1小時、5小時、24小時及48小時的時間點收集血漿樣本。使用ELISA測量各個抗體構築體之抗IL-13及抗TSLP抗原結合域的濃度。圖10所示之數據代表各個雙特異性抗體在每個時間點下兩隻動物的平均值。PX128雙特異性抗體顯示出有前景的體內穩定性：在構築體PX128-B、PX128-C及PX128-D中，抗TSLP抗原結合域濃度在長達48小時的所有測試時間點皆維持相對不變，且抗IL-13抗原結合域濃度在長達48小時的所有測試時間點皆維持相對不變。PX128-A 抗IL-13 scFv濃度隨時間推移而下降。實例4：靶向TSLP及IL-13的示例性雙特異性抗體：額外的配置。The PX128 IL-13 × TSLP bispecific antibodies described above were tested to assess average pharmacokinetic properties and validate the in vivo stability of each construct. Briefly, C57BL/6 mice were intravenously injected with a single 1 mg/kg dose of the bispecific antibody and divided into groups according to the bispecific antibody administered. Plasma samples were collected at 5 minutes, 1 hour, 5 hours, 24 hours, and 48 hours. The concentrations of the anti-IL-13 and anti-TSLP antigen-binding domains of each antibody construct were measured using ELISA. The data shown inFigure10 represent the average of two animals at each time point for each bispecific antibody. The PX128 bispecific antibody demonstrated promising in vivo stability: in constructs PX128-B, PX128-C, and PX128-D, the concentration of the anti-TSLP antigen-binding domain remained relatively constant at all time points tested, up to 48 hours, and the concentration of the anti-IL-13 antigen-binding domain remained relatively constant at all time points tested, up to 48 hours. The concentration of the PX128-A anti-IL-13 scFv decreased over time.Example4 :Exemplary bispecific antibodiestargetingTSLPandIL-13:Additional configurations.

設計並產生額外四種抗TSLP×抗IL-13雙特異性抗體配置，如圖11A至11D所示。圖11A顯示了PX128-I1設計配置的示意圖，其中該雙特異性抗體包含兩個各自融合(經由73P1 VH)至抗TSLP hz51B2-L3H9全長抗體重鏈之C端的抗IL-13 73P1 Fab。圖11B顯示了PX128-I2設計配置的示意圖，其中該雙特異性抗體包含兩個各自融合(經由hz51B2-L3H9 VH)至抗IL-13 73P1全長抗體重鏈之C端的抗TSLP hz51B2-L3H9 Fab。圖11C顯示了PX128-I3設計配置的示意圖，其中該雙特異性抗體包含兩個各自融合(經由73P1 Fab之CH1)至抗TSLP hz51B2-L3H9全長抗體之VH之N端的抗IL-13 73P1 Fab。圖11D顯示了PX128-I4設計配置的示意圖，其中該雙特異性抗體包含兩個各自融合(經由CH1 of hz51B2-L3H9 Fab)至抗IL-13 73P1全長抗體之VH之N端的抗TSLP hz51B2-L3H9 Fab。此等構築體之任一者可進一步包括CH1、CL及/或Fc域的突變，例如延長抗體半衰期的突變。Four additional anti-TSLP × anti-IL-13 bispecific antibody configurations were designed and generated, as shown inFigures11Ato11D .Figure11A shows a schematic diagram of the PX128-I1 design configuration, in which the bispecific antibody comprises two anti-IL-13 73P1 Fabs, each fused (via 73P1 VH) to the C-terminus of the anti-TSLP hz51B2-L3H9 full-length antibody heavy chain.Figure11B shows a schematic diagram of the PX128-I2 design configuration, in which the bispecific antibody comprises two anti-TSLP hz51B2-L3H9 Fabs, each fused (via hz51B2-L3H9 VH) to the C-terminus of the anti-IL-13 73P1 full-length antibody heavy chain.Figure11C shows a schematic diagram of the PX128-I3 design configuration, wherein the bispecific antibody comprises two anti-IL-13 73P1 Fabs, each fused (via the CH1 of the 73P1 Fab) to the N-terminus of the VH of the anti-TSLP hz51B2-L3H9 full-length antibody.Figure11D shows a schematic diagram of the PX128-I4 design configuration, wherein the bispecific antibody comprises two anti-TSLP hz51B2-L3H9 Fabs, each fused (via the CH1 of the hz51B2-L3H9 Fab) to the N-terminus of the VH of the anti-IL-13 73P1 full-length antibody. Any of these constructs may further include mutations in the CH1, CL, and/or Fc domains, such as mutations that extend the antibody half-life.

將編碼構築體PX128-I1、PX128-I2、PX128-I3及PX128-I4之多肽鏈的核酸轉染至HEK-293細胞中，以產生雙特異性分子的多肽並分泌至培養基中，如上面實例2所示。Nucleic acids encoding the polypeptide chains of constructs PX128-I1, PX128-I2, PX128-I3, and PX128-I4 were transfected into HEK-293 cells to produce bispecific polypeptides that were secreted into the culture medium, as described in Example 2 above.

PX128-I1 - PX128-I4 TSLP報導細胞試驗。同時將共同表現TSLPR/IL-7Rα的STAT5-螢光素酶HEK-293報導細胞與10 ng/mL人類TSLP及連續稀釋的雙特異性抗體之一者在37°C下培養5小時。同樣地，測試了TSLP×IL-13異二聚體雙特異性抗體形式PX128-R2 (圖13)，其包含抗TSLP hz51B2-L3H9結合域，且在抗IL-13 73P1結合域之CH1-CL配對中含有替代的雙硫鍵。在共同培養5小時之後，測量生物發光訊號。圖12A所示之數據顯示了螢光素酶的抑制百分比，作為抗體與TSLP分子結合並防止TSLP經由TSLPR/IL-7Rα受體複合體傳訊的讀數。此等結果證實，所有示例性IL-13×TSLP雙特異性抗體形式(PX128-I1-PX128-I4、PX128-R2)皆能有效阻斷TSLP傳訊，其中PX128-I2在較低濃度下表現出最高的TSLP抑制活性。此相當令人驚訝，係因在PX128-I2形式中抗TSLP Fab之VH域融合至Fc，其在TSLP結合中可具有空間抑制作用。PX128-I1 - PX128-I4 TSLPreporter cell assay . STAT5-luciferase HEK-293 reporter cells co-expressing TSLPR/IL-7Rα were incubated with 10 ng/mL human TSLP and serially diluted bispecific antibodies for 5 hours at 37°C. Similarly, the TSLP × IL-13 heterodimeric bispecific antibody format PX128-R2 (Figure13 ) was tested. It contains the anti-TSLP hz51B2-L3H9 binding domain and an alternative disulfide bond in the CH1-CL pairing of the anti-IL-13 73P1 binding domain. Bioluminescence signal was measured after 5 hours of co-incubation. The data presented inFigure12A show percent inhibition of luciferase, a measure of antibody binding to TSLP molecules and prevention of TSLP signaling through the TSLPR/IL-7Rα receptor complex. These results demonstrate that all exemplary IL-13×TSLP bispecific antibody formats (PX128-I1-PX128-I4, PX128-R2) effectively blocked TSLP signaling, with PX128-I2 exhibiting the highest TSLP inhibitory activity at lower concentrations. This is surprising, as the VH domain of the anti-TSLP Fab in the PX128-I2 format is fused to the Fc, which could have a steric inhibitory effect on TSLP binding.

PX128-I1 - PX128-I4 IL-13報導細胞試驗。將共同表現IL-13Rα1及IL-4Rα的SEAP HEK-293報導細胞株用於監控IL-13的傳訊路徑活性，其係藉由測量SEAP的產生且隨後分泌至條件培養基中。同時將HEK-293報導細胞與10 ng/mL人類IL-13及連續稀釋的雙特異性抗體在37°C下培養過夜。同樣地，測試了TSLP×IL-13異二聚體雙特異性抗體PX128-R2 (圖13)。將US-FDA/EMA批准的參考抗IL-13 Ab. #2、IgG形式的TSLP×IL-13雙特異性參考Ab. #3及VHH形式的TSLP×IL-13×HSA三特異性參考Ab. #4作為陽性對照。在共同培養之後，測量SEAP訊號。圖12B所示之數據顯示了SEAP產生及分泌的抑制百分比，作為抗體與IL-13分子結合並防止IL-13經由IL-13Rα1/IL-4Rα受體複合體傳訊的讀數。所有雙特異性抗體構築體(PX128-I1-PX128-I4、PX128-R2)皆表現出強的IL-13阻斷活性，類似於EMA批准的參考抗IL-13 Ab. #2。此外，所測試之TSLP×IL-13雙特異性抗體構築體的IL-13阻斷活性皆比參考TSLP×IL-13 Ab. #3及參考TSLP×IL-13×HSA Ab. #4的更高。實例5：異二聚體抗TSLP×抗IL-13雙特異性抗體。PX128-I1 - PX128-I4 IL-13reporter cell assay . A SEAP HEK-293 reporter cell line co-expressing IL-13Rα1 and IL-4Rα was used to monitor IL-13 signaling pathway activity by measuring SEAP production and subsequent secretion into conditioned medium. HEK-293 reporter cells were incubated with 10 ng/mL human IL-13 and serially diluted bispecific antibodies at 37°C overnight. Similarly, the TSLP × IL-13 heterodimer bispecific antibody PX128-R2 was tested (Figure13 ). US-FDA/EMA-approved reference anti-IL-13 Ab #2, TSLP × IL-13 bispecific reference Ab #3 in IgG format, and TSLP × IL-13 × HSA trispecific reference Ab #4 in VHH format served as positive controls. Following co-culture, SEAP signaling was measured. The data presented inFigure12B show the percent inhibition of SEAP production and secretion, a measure of antibody binding to IL-13 molecules and prevention of IL-13 signaling via the IL-13Rα1/IL-4Rα receptor complex. All bispecific antibody constructs (PX128-I1-PX128-I4, PX128-R2) exhibited potent IL-13 blocking activity, similar to that of the EMA-approved reference anti-IL-13 Ab #2. Furthermore, the IL-13 blocking activity of all tested TSLP × IL-13 bispecific antibody constructs was higher than that of reference TSLP × IL-13 Ab #3 and reference TSLP × IL-13 × HSA Ab #4.Example5: Heterodimeric anti-TSLP ×anti-IL-13bispecific antibody.

產生了具有各種CH1、CL及/或Fc域突變的異二聚體抗TSLP×抗IL-13雙特異性抗體。圖13顯示了IgG異二聚體雙特異性分子「PX128-R2」的示意圖，其包含(1)連接至具有臼突變(T366S、L368A及Y407V)之Fc域的抗TSLP hz51B2-L3H9 Fab片段，以及(2)含有連接至具有杵突變(T366W)之Fc域的輕鏈(人類λ CL)突變(E124C (λ)及C214S)及重鏈突變(F126C及C220S)的抗IL-13 73P1 Fab片段。各個突變皆以EU編號描述。杵臼突變可轉換各臂，亦即抗TSLP臂Fc上的杵及抗IL-13臂Fc上的臼。此異二聚體雙特異性抗體經由瞬時轉染在HEK-293細胞中表現。在蛋白A管柱上純化分泌至條件培養基中的IgG分子，隨後在陶瓷羥基磷灰石(CHT)或Capto MMC ImpRes的混合模式層析管柱上進一步純化。 Ÿ「R2突變」(λ)：HC (F126C及C220S)與LC (E124C及C214S)配對 Ÿ「R2突變」(κ)：HC (F126C及C220S)與LC (Q124C及C214S)配對Heterodimeric anti-TSLP × anti-IL-13 bispecific antibodies with various CH1, CL, and/or Fc domain mutations were generated.Figure13 shows a schematic diagram of the IgG heterodimeric bispecific molecule "PX128-R2," which comprises (1) an anti-TSLP hz51B2-L3H9 Fab fragment linked to an Fc domain with hole mutations (T366S, L368A, and Y407V), and (2) an anti-IL-13 73P1 Fab fragment containing light chain (human λ CL) mutations (E124C (λ) and C214S) and heavy chain mutations (F126C and C220S) linked to an Fc domain with a knob mutation (T366W). Each mutation is described by EU numbering. The knob-to-hole mutation converts each arm, i.e., the knob on the anti-TSLP arm Fc and the hole on the anti-IL-13 arm Fc. This heterodimeric bispecific antibody was expressed in HEK-293 cells by transient transfection. IgG molecules secreted into conditioned medium were purified on a protein A column and further purified on a ceramic hydroxyapatite (CHT) or Capto MMC ImpRes mixed-mode chromatography column. Ÿ"R2 mutation" (λ): HC (F126C and C220S) paired with LC (E124C and C214S) Ÿ"R2 mutation" (κ): HC (F126C and C220S) paired with LC (Q124C and C214S)

如圖14所示，產生了第二及第三異二聚體抗TSLP×抗IL-13雙特異性IgG 抗體，其中該異二聚體雙特異性抗體包括(1)連接至具有臼突變(T366S、L368A及Y407V)之Fc域的抗TSLP hz51B2-L3H9 Fab，以及(2)含有連接至具有杵突變(T366W)之Fc域的輕鏈突變(L135F)及重鏈突變(「JT11」：F170V、S183I及V185L；或「JT7」：F170I、S183L及V185L)的抗IL-13 73P1 Fab片段。各個突變皆以EU編號描述。杵臼突變可轉換各臂，亦即抗TSLP臂Fc上的杵及抗IL-13臂Fc上的臼。此等異二聚體雙特異性抗體「PX128-JT11」及「PX128-JT7」分別依上述方法產生及純化。 Ÿ「JT11突變」：HC (F170V、S183I及V185L)與LC L135F配對 Ÿ「JT7突變」：HC (F170I、S183L及V185L)與LC L135F配對As shownin FIG14, a second and third heterodimeric anti-TSLP × anti-IL-13 bispecific IgG antibody was generated, wherein the heterodimeric bispecific antibody comprises (1) anti-TSLP hz51B2-L3H9 Fab linked to an Fc domain having hole mutations (T366S, L368A, and Y407V), and (2) anti-IL-13 73P1 Fab fragment containing a light chain mutation (L135F) and heavy chain mutations ("JT11": F170V, S183I, and V185L; or "JT7": F170I, S183L, and V185L) linked to an Fc domain having a knob mutation (T366W). Each mutation is described by EU numbering. Knob-to-hole mutations convert the Fc knob on the anti-TSLP arm and the hole on the anti-IL-13 arm. These heterodimeric bispecific antibodies, "PX128-JT11" and "PX128-JT7," were generated and purified using the methods described above. Ÿ"JT11 mutation": HC (F170V, S183I, and V185L) paired with LC L135F Ÿ"JT7 mutation": HC (F170I, S183L, and V185L) paired with LC L135F

如圖15所示，產生了第四及第五異二聚體抗TSLP×抗IL-13雙特異性IgG 抗體，其中該異二聚體雙特異性抗體包括(1)連接至具有臼突變(T366S、L368A及Y407V)之Fc域的抗TSLP hz51B2-L3H9 Fab，以及(2)含有連接至具有杵突變(T366W)之Fc域的輕鏈突變(E124C (λ)、L135F及C214S)及重鏈突變(JT11/R2：F126C、F170V、S183I、V185L及C220S；或JT7/R2：F126C、F170I、S183L、V185L及C220S)的抗IL-13 73P1 Fab片段。各個突變皆以EU編號描述。杵臼突變可轉換各臂，亦即抗TSLP臂Fc上的杵及抗IL-13臂Fc上的臼。此等異二聚體雙特異性抗體「PX128-JT11/R2」及「PX128-JT7/R2」分別依上述方法產生及純化。 Ÿ「JT11/R2突變」: HC (F126C、F170V、S183I、V185L及C220S)與LC (E124C、L135F及C214S)配對 Ÿ「JT7/R2突變」: HC (F126C、F170I、S183L、V185L及C220S)與LC (E124C、L135F及C214S)配對As shownin FIG15, the fourth and fifth heterodimeric anti-TSLP×anti-IL-13 bispecific IgG antibodies were generated, wherein the heterodimeric bispecific antibodies comprised (1) anti-TSLP hz51B2-L3H9 Fab linked to an Fc domain with hole mutations (T366S, L368A, and Y407V), and (2) anti-TSLP hz51B2-L3H9 Fab containing a light chain mutation (E124C) linked to an Fc domain with a knob mutation (T366W). The anti-IL-13 73P1 Fab fragment was constructed with two heavy chain mutations (JT11/R2: F126C, F170V, S183I, V185L, and C220S; or JT7/R2: F126C, F170I, S183L, V185L, and C220S). Each mutation is represented by the EU numbering. The knob-to-hole mutations convert the knob on the anti-TSLP arm and the hole on the anti-IL-13 arm. These heterodimeric bispecific antibodies, "PX128-JT11/R2" and "PX128-JT7/R2," were generated and purified, respectively, using the methods described above. Ÿ"JT11/R2 mutation": HC (F126C, F170V, S183I, V185L, and C220S) paired with LC (E124C, L135F, and C214S) Ÿ"JT7/R2 mutation": HC (F126C, F170I, S183L, V185L, and C220S) paired with LC (E124C, L135F, and C214S)

測試了PX128-JT11及PX128-JT7異二聚體雙特異性抗體，以評估體內平均藥物動力學特徵並驗證各個構築體的體內穩定性。簡言之，C57BL/6小鼠以1 mg/kg的單一劑量PX128-JT11或PX128-JT7進行靜脈內注射，並根據投予的雙特異性抗體進行分組。在0小時、24小時、48小時、72小時、96小時、120小時、144小時及168小時的時間點收集血漿樣本。使用ELISA測量各個雙特異性抗體之抗IL-13及抗TSLP抗原結合域的濃度。圖16所示之數據代表各個雙特異性抗體在每個時間點下兩隻動物的平均值。圖16證實了PX128-JT11及PX128-JT7雙特異性抗體顯示出有前景的體內穩定性，係因各個抗原結合域的濃度在投予後長達168小時內維持驚人的穩定性且濃度下降最少。此特別高的穩定性可允許以高濃度(例如，＞100 mg/ml)調配雙特異性抗體，從而使其適合各種形式的投予，例如皮下投予。PX128-JT11 and PX128-JT7 heterodimeric bispecific antibodies were tested to assess the average in vivo pharmacokinetic properties and validate the in vivo stability of each construct. Briefly, C57BL/6 mice were intravenously injected with a single 1 mg/kg dose of PX128-JT11 or PX128-JT7 and divided into groups according to the bispecific antibody administered. Plasma samples were collected at 0, 24, 48, 72, 96, 120, 144, and 168 hours. The concentrations of the anti-IL-13 and anti-TSLP antigen-binding domains of each bispecific antibody were measured using ELISA. The data presented inFigure16 represent the average of two animals at each time point for each bispecific antibody.Figure16 demonstrates that the PX128-JT11 and PX128-JT7 bispecific antibodies exhibit promising in vivo stability, as the concentration of each antigen-binding domain remains remarkably stable with minimal decrease in concentration for up to 168 hours after administration. This exceptionally high stability allows the bispecific antibodies to be formulated at high concentrations (e.g., >100 mg/ml), making them suitable for various modes of administration, such as subcutaneous administration.

異二聚體IgG雙特異性抗體與R2、JT7、JT11、JT7/R2或JT11/R2突變阻斷TSLP的比較：測試了上述各個異二聚體IgG雙特異性抗體在HEK-293報導細胞中阻斷人類TSLP與TSLPR/IL-7Rα複合體結合的能力。抗IL-13臂的親代CL域(不含突變)可含有SEQ ID NO: 74 (「CL λ L2 wt」形式)或75 (「CL λ L1 wt」形式)之序列，其等為CL λ 同型。基於CL λ L1 wt形式產生「PX128-R2」突變。當基於CL λ L2 wt形式產生R2突變時，所產生的雙特異性抗體稱為「PX128-R2L2」。同時將共同表現TSLPR/IL-7Ra的STAT5-螢光素酶HEK-293報導細胞與10 ng/ml人類TSLP及連續稀釋的各個異二聚體雙特異性抗體在37°C下培養5小時。在共同培養5小時之後，測量生物發光訊號。圖17A之數據顯示了螢光素酶的抑制百分比，包括計算的IC90值。此等結果證實，所有測試的示例性異二聚體IL-13×TSLP雙特異性抗體形式皆能有效阻斷游離TSLP與TSLP受體複合體的結合並表現出類似的IC90特徵，其中PX128-R2及PX128-JT11表現出最高的TSLP抑制活性。Comparison ofTSLPBlockade byHeterodimericIgGBispecific Antibodies withR2,JT7,JT11,JT7/R2, orJT11/R2 Mutations: Each of the heterodimeric IgG bispecific antibodies was tested for its ability to block binding of human TSLP to the TSLPR/IL-7Rα complex in HEK-293 reporter cells. The parental CL domain (lacking mutations) of the anti-IL-13 arm can contain the sequence of SEQ ID NO: 74 ("CLλL2 wt" form) or 75 ("CLλL1 wt" form), which are CLλ isotypes. The "PX128-R2" mutation was generated based on the CLλL1 wt form. When the R2 mutation is generated based on the wt form of CLλL2, the resulting bispecific antibody is designated "PX128-R2L2." HEK-293 reporter cells expressing STAT5-luciferase and co-expressing TSLPR/IL-7Ra were incubated with 10 ng/ml human TSLP and serially diluted heterodimeric bispecific antibodies for 5 hours at 37°C. After 5 hours of co-incubation, bioluminescence signals were measured. Data inFigure17A show the percent inhibition of luciferase, including calculatedIC90 values. These results demonstrate that all tested exemplary heterodimeric IL-13×TSLP bispecific antibody formats can effectively block the binding of free TSLP to the TSLP receptor complex and exhibit similarIC90 characteristics, with PX128-R2 and PX128-JT11 exhibiting the highest TSLP inhibitory activity.

異二聚體IgG雙特異性抗體與R2、JT7、JT11、JT7/R2或JT11/R2突變阻斷IL-13的比較：測試了上述各個異二聚體IgG雙特異性抗體在HEK-293-SEAP報導細胞中阻斷人類IL-13與IL-13Rα1/IL-4Rα受體複合體結合的能力。同時將HEK-293-SEAP報導細胞與10 ng/mL人類IL-13及連續稀釋的各個異二聚體雙特異性抗體在37°C下培養過夜。在共同培養之後，測量SEAP訊號。圖17B所示之數據顯示了SEAP產生及分泌的抑制百分比，作為抗體與IL-13分子結合並防止IL-13與IL-13Rα1/IL-4Rα受體複合體結合的讀數。此等結果證實，所有測試的示例性異二聚體IL-13×TSLP雙特異性抗體形式皆能有效阻斷游離IL-13與IL-13Rα1/IL-4Rα受體複合體的結合並表現出類似的IC90特徵。Comparison ofIL-13Blockade byHeterodimericIgGBispecific Antibodies withR2,JT7,JT11,JT7/R2, orJT11/R2 Mutations: Each of the heterodimeric IgG bispecific antibodies was tested for its ability to block the binding of human IL-13 to the IL-13Rα1/IL-4Rα receptor complex in HEK-293-SEAP reporter cells. HEK-293-SEAP reporter cells were incubated overnight at 37°C with 10 ng/mL human IL-13 and serially diluted versions of each heterodimeric bispecific antibody. Following co-incubation, SEAP signaling was measured. The data presented inFigure17B show the percent inhibition of SEAP production and secretion, as a readout of antibody binding to the IL-13 molecule and preventing IL-13 from binding to the IL-13Rα1/IL-4Rα receptor complex. These results demonstrate that all exemplary heterodimeric IL-13×TSLP bispecific antibody formats tested effectively blocked the binding of free IL-13 to the IL-13Rα1/IL-4Rα receptor complex and exhibited similarIC90 characteristics.

基於TSLP初代細胞之試驗：藉由評估人類PBMC中TSLP誘導的CCL17產生水平來測試PX128-JT11及PX128-JT7異二聚體雙特異性抗體的劑量抑制反應。將健康供體PBMC與0.5 ng/mL重組TSLP及連續稀釋的PX128-JT11或PX128-JT7異二聚體雙特異性抗體共同培養36小時。將US-FDA批准的參考抗TSLP Ab. #5及IgG形式的TSLP×IL-13雙特異性參考Ab. #3作為對照。藉由CCL17 ELISA套組來測量新鮮收集上清液中的CCL17濃度。使用Graphpad Prism計算IC90值。圖18A顯示了與US-FDA批准的參考抗TSLP Ab. #5及參考TSLP×IL-13 Ab. #3相比，PX128-JT11及PX128-JT7表現出類似的或可能略強的TSLP傳訊抑制作用。TSLP-basedprimary cell assay : The inhibitory effects of PX128-JT11 and PX128-JT7 heterodimeric bispecific antibodies were tested by assessing TSLP-induced CCL17 production in human PBMCs. PBMCs from healthy donors were incubated with 0.5 ng/mL recombinant TSLP and serially diluted PX128-JT11 or PX128-JT7 heterodimeric bispecific antibodies for 36 hours. US-FDA-approved reference anti-TSLP Ab #5 and IgG-based TSLP × IL-13 bispecific reference Ab #3 were used as controls. CCL17 concentrations were measured in freshly collected supernatants using a CCL17 ELISA kit.IC90 values were calculated using Graphpad Prism.Figure18A shows that PX128-JT11 and PX128-JT7 exhibited similar or possibly slightly stronger inhibition of TSLP signaling compared to the US-FDA-approved reference anti-TSLP Ab #5 and reference TSLP×IL-13 Ab #3.

基於IL-13初代細胞之試驗：藉由評估人類PBMC中IL-13誘導的CCL17產生水平來測試PX128-JT11及PX128-JT7異二聚體雙特異性抗體的劑量抑制反應。將健康供體PBMC與5 ng/mL重組IL-13及連續稀釋的PX128-JT11或PX128-JT7異二聚體雙特異性抗體共同培養36小時。將EMA批准的參考抗IL-13 Ab. #2及IgG形式的TSLP×IL-13雙特異性參考Ab. #3作為對照。藉由CCL17 ELISA套組來測量新鮮收集上清液中的CCL17濃度。使用Graphpad Prism計算IC90值。圖18B顯示了與EMA批准的參考抗IL-13 Ab. #2及參考Ab. #3相比，PX128-JT11及PX128-JT7表現出類似地強的IL-13傳訊抑制作用(皮莫耳範圍)。PrimaryIL-13 cell-based assay: The inhibitory effects of PX128-JT11 and PX128-JT7 heterodimeric bispecific antibodies were assessed by assessing IL-13-induced CCL17 production in human PBMCs. PBMCs from healthy donors were incubated with 5 ng/mL recombinant IL-13 and serially diluted PX128-JT11 or PX128-JT7 heterodimeric bispecific antibodies for 36 hours. EMA-approved reference anti-IL-13 Ab #2 and TSLP × IL-13 bispecific Ab #3 in IgG format served as controls. CCL17 concentrations were measured in freshly collected supernatants using a CCL17 ELISA kit.IC90 values were calculated using Graphpad Prism.Figure18B shows that PX128-JT11 and PX128-JT7 exhibited similarly potent inhibition of IL-13 signaling (in the picomolar range) compared to EMA-approved reference anti-IL-13 Ab #2 and Ab #3.

基於TSLP/IL-13組合初代細胞之試驗：藉由評估人類PBMC中TSLP/IL-13組合誘導的CCL17產生水平來測試PX128-JT11及PX128-JT7異二聚體雙特異性抗體的劑量抑制反應。將健康供體PBMC與0.5 ng/mL TSLP和5 ng/mL IL-13及連續稀釋的PX128-JT11或PX128-JT7異二聚體雙特異性抗體共同培養36小時。將IgG形式的TSLP×IL-13雙特異性參考Ab. #3及EMA批准的參考抗IL-13 Ab. #2 + US-FDA批准的參考抗TSLP Ab. #5之組合作為陽性對照。藉由CCL17 ELISA套組來測量新鮮收集上清液中的CCL17濃度。使用Graphpad Prism計算IC90值。圖18C顯示了與US-FDA/EMA批准的參考Ab. #2及#5之組合相比，PX128-JT11及PX128-JT7表現出類似地強的IL-13及TSLP傳訊共同抑制作用，且與參考TSLP×IL-13 Ab. #3相比表現出IL-13及TSLP傳訊的抑制作用有所改善。TSLP/IL-13Combination Primary Cell-Based Assay: The inhibitory effects of the PX128-JT11 and PX128-JT7 heterodimeric bispecific antibodies were assessed by evaluating the TSLP/IL-13 combination-induced CCL17 production in human PBMCs. PBMCs from healthy donors were incubated with 0.5 ng/mL TSLP and 5 ng/mL IL-13 and serially diluted PX128-JT11 or PX128-JT7 heterodimeric bispecific antibodies for 36 hours. The IgG-based TSLP × IL-13 bispecific reference Ab #3 and the combination of the EMA-approved reference anti-IL-13 Ab #2 and the US-FDA-approved reference anti-TSLP Ab #5 served as positive controls. CCL17 concentrations in freshly collected supernatants were measured using a CCL17 ELISA kit.IC90 values were calculated using GraphPad Prism.Figure18C shows that PX128-JT11 and PX128-JT7 exhibited similarly potent co-inhibition of IL-13 and TSLP signaling compared to the US-FDA/EMA-approved reference Ab #2 and #5 combination, and demonstrated improved inhibition of both IL-13 and TSLP signaling compared to the reference TSLP × IL-13 Ab #3.

TSLP×IL-13 BsAb藥物動力學測定：藉由皮下注射或靜脈內推注至食蟹獼猴(「cyno」)中來測試示例性PX128-JT11異二聚雙特異性抗體的體內半衰期，如下表10所示。根據最近的體重計算個體劑量。將參考TSLP×IL-13×HSA Ab. #4作為對照。表10.TSLP×IL-13抗體給藥。研究設計組編號測試材料投予途徑劑量水平(mg/kg)劑量體積a(mL/kg)劑量濃度(mg/mL)動物數量1PX128-JT11SC1011023PX128-JT11IV1011023參考Ab. #4IV101102a根據最近的體重計算個體劑量體積。 IV：靜脈內；SC：皮下TSLP × IL-13 BsAbPharmacokinetics : The in vivo half-life of the exemplary PX128-JT11 heterodimeric bispecific antibody was determined in cynomolgus macaques ("cyno") by subcutaneous injection or intravenous bolus injection, as shown inTable10 below. Individual doses were calculated based on the most recent body weight. A reference TSLP × IL-13 × HSA Ab #4 was used as a control.Table10. TSLP × IL-13 Antibody Dosing.Research Design Group No.Test materialsInvestment channelsDosage level(mg/kg)Dose volumea(mL/kg)Dosage concentration(mg/mL)Number of animals 1 PX128-JT11 SC 10 1 10 2 3 PX128-JT11 IV 10 1 10 2 3 See Ab. #4 IV 10 1 10 2aIndividual dose volume is calculated based on the most recent body weight. IV: intravenous; SC: subcutaneous

從靜脈內推注(i.v.)投予PX128-JT11或參考TSLP×IL-13×HSA Ab. #4的​​食蟹獼猴中收集血液樣本，其係於給藥之前(亦即，給藥前)及給藥之後的以下時間點進行：15分鐘、1小時、2小時、6小時、1天、2天、3天、5天、7天、10天、14天、21天、28天、42天、65天及70天。從皮下注射(s.c.)投予PX128-JT11的​​食蟹獼猴中收集血液樣本，其係於給藥之前(亦即，給藥前)及給藥之後的以下時間點進行：15分鐘、1小時、2小時、6小時、1天、2天、3天、5天、7天、10天、14天、21天、28天、42天、65天及70天。Blood samples were collected from cynomolgus macaques that received either PX128-JT11 or the reference TSLP×IL-13×HSA Ab. #4 by intravenous bolus (i.v.) at 15 minutes, 1 hour, 2 hours, 6 hours, 1 day, 2 days, 3 days, 5 days, 7 days, 10 days, 14 days, 21 days, 28 days, 42 days, 65 days, and 70 days after dosing. Blood samples were collected from cynomolgus macaques administered PX128-JT11 subcutaneously (s.c.) at 15 minutes, 1 hour, 2 hours, 6 hours, 1 day, 2 days, 3 days, 5 days, 7 days, 10 days, 14 days, 21 days, 28 days, 42 days, 65 days, and 70 days after administration.

從血液樣本中分離血清，隨後使用合格的基於Mesoscale Discovery (MSD)的夾層法來測量各個示例性PX128-JT11雙特異性抗體及參考TSLP×IL-13×HSA Ab. #4的​​濃度。在食蟹獼猴血清中的定量下限(LLOQ)確定為約4 ng/mL。Serum was separated from the blood samples, and the concentrations of each exemplary PX128-JT11 bispecific antibody and reference TSLP × IL-13 × HSA Ab #4 were measured using a qualified Mesoscale Discovery (MSD)-based sandwich assay. The lower limit of quantification (LLOQ) in cynomolgus macaque serum was determined to be approximately 4 ng/mL.

使用了在Phoenix® 64 WinNonlin® 8.4版軟體(Certara，Princeton，NJ，USA)中執行的非隔間分析(NCA)來評估示例性PX128-JT11 TSLP×IL-13雙特異性抗體及參考TSLP×IL-13×HSA Ab. #4的藥物動力學(PK)參數。針對s.c.組的個體血清濃度數據，採用血管外投予途徑的非隔間分析。針對i.v.組的個體血清濃度數據，採用i.v.推注投予途徑的非隔間分析。Pharmacokinetic (PK) parameters of the exemplary PX128-JT11 TSLP × IL-13 bispecific antibody and reference TSLP × IL-13 × HSA Ab #4 were evaluated using non-compartmental analysis (NCA) performed in Phoenix® 64 WinNonlin® version 8.4 software (Certara, Princeton, NJ, USA). Individual serum concentration data for the s.c. group were analyzed using non-compartmental analysis for the extravascular route of administration. Individual serum concentration data for the i.v. group were analyzed using non-compartmental analysis for the i.v. bolus route of administration.

如圖19所示，在向食蟹獼猴靜脈內推注單一劑量之後，示例性PX128-JT11的體內半衰期為約26天，而參考TSLP×IL-13×HSA Ab. #4的體內半衰期為約4.75天。在皮下注射單一劑量的示例性PX128-JT11之後，體內半衰期亦隨時間顯著高於靜脈推注的參考TSLP×IL-13×HSA Ab. #4。As shown inFigure19 , after a single intravenous bolus dose in cynomolgus macaques, the in vivo half-life of exemplary PX128-JT11 was approximately 26 days, while the in vivo half-life of reference TSLP × IL-13 × HSA Ab. #4 was approximately 4.75 days. Following a single subcutaneous dose of exemplary PX128-JT11, the in vivo half-life was also significantly higher over time than that of reference TSLP × IL-13 × HSA Ab. #4, which was administered as an intravenous bolus.

綜上所述，上述結果證實，與US-FDA批准的參考抗TSLP Ab. #5相比，示例性雙特異性抗體針對TSLP的抑制活性類似或可能略強，且與US-FDA/EMA批准的參考抗IL-13 Ab. #2、參考TSLP×IL-13 Ab. #3及參考TSLP×IL-13×HSA Ab. #4相比，針對IL-13的抑制活性類似或有所改善。特別地，與EMA批准的第II類抗IL-13參考Ab. #2及US-FDA批准的第I類抗IL-13參考Ab. #1相比，抗IL-13 73P1殖株顯示出更有效地阻斷IL-13傳訊。參考抗IL-13 Ab. #1 (第I類)及#2 (第II類)的臨床試驗證實，更好的IL-13阻斷活性與發炎性疾病(如異位性皮膚炎)的臨床功效改善一致。此外，結果表明，與參考TSLP×IL-13×HSA Ab. #4相比，非人類靈長類動物的體內半衰期顯著改善。總之，此等結果表明，本文所述之示例性抗IL-13抗體、示例性抗TSLP抗體及所有包含其之示例性雙特異性抗體針對發炎性疾病(諸如異位性皮膚炎、COPD及氣喘)將具有臨床效果，可能顯示出比現有更好的治療功效。In summary, the above results demonstrate that the exemplary bispecific antibodies exhibit similar or potentially slightly stronger inhibitory activity against TSLP compared to the US-FDA-approved reference anti-TSLP Ab. #5, and similar or improved inhibitory activity against IL-13 compared to the US-FDA/EMA-approved reference anti-IL-13 Ab. #2, reference TSLP×IL-13 Ab. #3, and reference TSLP×IL-13×HSA Ab. #4. In particular, the anti-IL-13 73P1 clone demonstrated greater efficacy in blocking IL-13 signaling compared to the EMA-approved Class II anti-IL-13 reference Ab. #2 and the US-FDA-approved Class I anti-IL-13 reference Ab. #1. Clinical trials of reference anti-IL-13 Abs #1 (Class I) and #2 (Class II) demonstrated that enhanced IL-13 blocking activity correlated with improved clinical efficacy in inflammatory diseases such as atopic dermatitis. Furthermore, results demonstrated significantly improved in vivo half-life in non-human primates compared to reference TSLP × IL-13 × HSA Ab #4. Taken together, these results suggest that the exemplary anti-IL-13 antibodies, exemplary anti-TSLP antibodies, and all exemplary bispecific antibodies described herein will be clinically effective against inflammatory diseases such as atopic dermatitis, COPD, and asthma, potentially demonstrating improved therapeutic efficacy compared to currently available options.

本發明中提及的所有參考文獻皆通過引用併入本文，如同每一參考文獻皆通過引用單獨併入本文一般。儘管該描述參考特定實施例，但本領域技術人員應明瞭，本發明可利用此等具體細節的變化來實踐。因此，本發明不應理解為限於本文所述之實施例。 序列表SEQ ID NO:描述序列1抗TSLP 51A1 VH (CDR加底線)QVQLQQSGAELVRPGASVKLSCKASGYMFTDYAMHWVKQTPVHGLEWIGTIDPETGGTAYTQKFKGKATLTADISSSTAYVELRSLTSEDSAVYYCTREGSYGGYVDYWGQGTTLSVSS2抗TSLP 51A1/51A4 CDR-H1DYAMH3抗TSLP 51A1/51A4 CDR-H2TIDPETGGTAYTQKFKG4抗TSLP 51A1/51A3/51A4/ 51A6 CDR-H3EGSYGGYVDY5抗TSLP 51A1VL (CDR加底線)DIVMTQSQKFMSTSVGDRVNITCKASQNVHTAVAWYQQKPGQSPKTLIFLASNRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCLQLWNYPYTFGGETKLEIK6抗TSLP 51A1/51A3/51A4/51A6 CDR-L1KASQNVHTAVA7抗TSLP 51A1/51A3/51A4/ 51A6 CDR-L2LASNRHT8抗TSLP 51A1/51A3/51A4/51A6 CDR-L3LQLWNYPYT9抗TSLP 51A4 VH (CDR加底線)QVQLQQSGAELVRPGASVKLSCKASGYMFTDYAMHWVKQTPVHGLEWIGTIDPETGGTAYTQKFKGKATLTADISSNTAYVELRSLTSEDSAVYYCTREGSYGGYVDYWGQGTTLSVSS10抗TSLP 51A4 VL (CDR加底線)DIVMTQSQKFMSTSVGDRVSITCKASQNVHTAVAWYQQKPGQSPKTLIFLASNRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCLQLWNYPYTFGGGTKLEIK11抗TSLP 51A3/51A6 VH  (CDR加底線)QVQLQQSGAELVRPGASVKLSCKASGYTFTDYAMHWVKQTLVHGLEWVGTIDPETGGTAYTQKFQGKATLTADVSSSTAYVELRSLTSEDSAVYYCTREGSYGGYVDYWGQGTTLSVSS12抗TSLP 51A3/51A6 CDR-H1DYAMH13抗TSLP 51A3/51A6 CDR-H2TIDPETGGTAYTQKFQG14連接子，其中n為至少1的整數(GSGGS)n15抗TSLP 51A6 VL 抗TSLP 51A3 VL (CDR加底線)DIVMTQSQKFMSTSEGDRVSITCKASQNVHTAVAWYQQKPGQSPKTLIFLASNRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCLQLWNYPYTFGGGTKLEIK16連接子，其中n為至少1的整數(GGGS)n17連接子，其中n為至少1的整數(GGGGS)n18連接子GGSG19抗TSLP 51B2 VH (CDR加底線)QVQLLQPGTELVKPGASVKLSCKASGYTFTSYLMHWVKQRPGQGLEWIGEIDPSDSYTDYNQNFRGKATLTVDMSSSTAYMQLSSLTSEDSAVYYCALTGTINYFDYWGQGTTLTVSS20抗TSLP 51B2 CDR-H1SYLMH21抗TSLP 51B2 CDR-H2EIDPSDSYTDYNQNFRG22抗TSLP 51B2 CDR-H3TGTINYFDY23抗TSLP 51B2 VL (CDR加底線)DIVMTQSHKFMSTSVGDRVNITCKASQDVVTAVAWYQQKPGQSPKLLIYWASMRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSSFTFTFGSGTKLEIK24抗TSLP 51B2 CDR-L1KASQDVVTAVA25抗TSLP 51B2 CDR-L2WASMRHT26抗TSLP 51B2 CDR-L3QQYSSFTFT27抗TSLP 54A2 VH (CDR加底線)QVQLRESGPGLVAPSQSLSITCTVSGFSLTSYGVNWVRQPPGKGLEWLGVIWDDGTTNYHSAFISRLSITKDNSKSQVFLKLNSLQTDDTATYYCAKGSIYWYFDVWGAGTTVTVSS28抗TSLP 54A2 CDR-H1SYGVN29抗TSLP 54A2 CDR-H2VIWDDGTTNYHSAFIS30抗TSLP 54A2 CDR-H3GSIYWYFDV31抗TSLP 54A2 VL (CDR加底線)DIVLTQSPASLAVSLGQRATISCKASQSVDYEGDSHMNWYQQKPGQSPKLLIYVASNLESGIPARFSGSGSGTDFTLNIHPVEEEDAATYYCQQSNEVPFTFGSGTKLEIK32抗TSLP 54A2 CDR-L1KASQSVDYEGDSHMN33抗TSLP 54A2 CDR-L2VASNLES34抗TSLP 54A2 CDR-L3QQSNEVPFT35抗TSLP 54B3 VH (CDR加底線)QVQLQQSGAELVRPGASVKLSCKASGYTFTDYEIHWVRQTPVHGLEWIGLIDPETGGTAHNQKFKAKATLTVDKSSTTAYMELRSLTSEDSAVYYCTREGQLGLSLDYWGQGTTLTVSS36抗TSLP 54B3 CDR-H1DYEIH37抗TSLP 54B3 CDR-H2LIDPETGGTAHNQKFKA38抗TSLP 54B3 CDR-H3EGQLGLSLDY39抗TSLP 54B3 VL (CDR加底線)DIVMTQSHKFMSTSVGERVSITCKASQDVGTTVAWYQQKPGQSPKLLIYWTSTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSTYPLTFGSGTKLEIQ40抗TSLP 54B3 CDR-L1KASQDVGTTVA41抗TSLP 54B3 CDR-L2WTSTRHT42抗TSLP 54B3 CDR-L3QQYSTYPLT43抗TSLP 54C4 VH (CDR加底線)EVQLQQSGPVLVKPGASVKMSCKASGYTFTDYYMNWVKQSHGKSLEWIGVINPYNADTKFNQRFKDKATLTVDESSSTAYMELNSLTSEDSAVYYCARGDDYAEESFAYWGQGTLVTVSAA44抗TSLP 54C4 CDR-H1DYYMN45抗TSLP 54C4 CDR-H2VINPYNADTKFNQRFKD46抗TSLP 54C4 CDR-H3GDDYAEESFAY47抗TSLP 54C4 VL (CDR加底線)QIVLTQSPSIMSASLGEEITLTCSASSSVSYMHWYQQKSGTSPKLLIYFTSNLASGVPSRFSGSGSGTFYSLTISSVEAEDAADYYCHQWSSYPTFGGGTKLEIK48抗TSLP 54C4 CDR-L1SASSSVSYMH49抗TSLP 54C4 CDR-L2FTSNLAS50抗TSLP 54C4 CDR-L3HQWSSYPT51抗TSLP hz51B2 VL-1 (CDR加底線)DIQLTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIK52抗TSLP hz51B2 VL-2 (CDR加底線)DIQLTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGQGTKLEIK53抗TSLP hz51B2 VL-3 (CDR加底線)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIK54抗TSLP hz51B2 VL-4 (CDR加底線)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGQGTKLEIK55抗TSLP hz51B2 VH-1QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWMGEIDPSDSYTDYNQNFRGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS56抗TSLP hz51B2 VH-2QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWMGEIDPSDSYTDYNQNFRGRVTMTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS57抗TSLP hz51B2 VH-3QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWIGEIDPSDSYTDYNQNFRGRVTMTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS58抗TSLP hz51B2 VH-4QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWMGEIDPSDSYTDYNQNFRGRVTLTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS59抗TSLP hz51B2 VH-5QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWIGEIDPSDSYTDYNQNFRGRVTLTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS60抗TSLP hz51B2 VH-6QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWMGEIDPSDSYTDYNQNFRGRATMTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS61抗TSLP hz51B2 VH-7QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWIGEIDPSDSYTDYNQNFRGRATMTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS62抗TSLP hz51B2 VH-8QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQGLEWIGEIDPSDSYTDYNQNFRGRATLTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS63抗TSLP hz51B2 VH-9QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS64抗TSLP hz51B2 VH-10QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWIGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS65抗IL-13 73P1 VH (CDR加底線)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS66抗IL-13 73P1 CDR-H1SYAMS67抗IL-13 73P1 CDR-H2AFSGSGGSTYYADSVKG68抗IL-13 73P1 CDR-H3DGLGPYFYNYGMDV69抗IL-13 73P1 VL (CDR加底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL70抗IL-13 73P1 CDR-L1SGDKLGDKYAY71抗IL-13 73P1 CDR-L2HDSKRPS72抗IL-13 73P1 CDR-L3QAWDSSTYV73CL κ wtRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC74CL λ L2 wtGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS75CL λ L1 wtGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS76人類IgG1 FcDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK77IgG1 Fc-LALADKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK78IgG1 Fc-LALA-LSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK79IgG1 Fc-LALA-YTEDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK80IgG1 Fc-LALA-LS-臼DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK81IgG1 Fc-LALA-LS-杵DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK82人類IgG1ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*83IgG1 LALAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK84IgG1 LALA-LSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK85IgG1 LALA-LS-杵ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK86IgG1 LALA-LS-臼ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK87IgG1 LALA-LS-臼 – JT7ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK88IgG1 LALA-LS-杵 – JT7ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK89IgG1 LALA-LS-臼 – JT11ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK90IgG1 LALA-LS-杵 – JT11ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK91IgG1 LALA-LS-臼 – JT7/R2ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK92IgG1 LALA-LS-杵 – JT7/R2ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK93IgG1 LALA-LS-臼 – JT11/R2ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK94IgG1 LALA-LS-杵 – JT11/R2ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK95IgG1 Fc-LALA-臼DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK96IgG1 Fc-LALA-杵DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK97連接子PLGLAG98連接子 1GGGG99連接子 2GGGGSGGGGSGGGGS101抗IL-13 73P1-HC-LALA (VH加斜體及底線)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK102抗IL-13 73P1-LC.C34Y (CL λ L1 wt) (VL加斜體及底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS103抗TSLP hz51B2-LC3 (CL κ wt) (VL加斜體及底線)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC104抗TSLP hz51B2-HC9 (IgG1 LALA) (VH加斜體及底線)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK105抗TSLP hz51B2-HC9 (IgG1 LALA-YTE) (VH加斜體及底線)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK106抗TSLP hz51B2-HC9LC3 scFv-stab (連接子加粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGCGTKLEIK107抗TSLP hz51B2-HC9LC4 scFv-stab (連接子加粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK108抗IL-13 73P1 scFv-LH (連接子加粗體)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS109抗TSLP hz51B2-HC9 VH-CH1 (VH加斜體及底線)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC110(hz51B2-H9-CH1)-(73P1_scFv-LH)-Fc.LALA.YTE (hz51B2-H9-VH加斜體及底線；連接子加粗體；73P1-VH加底線；73P1-VL加斜體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSGGGGDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 11173P1-Fc.LALA.YTE-hz51B2 H9L4 scFv HL-stab (連接子加粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK11273P1HC-hz51B2.H9L4 scFv HL-stab-Fc.LALA.YTEEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCGGGGQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIKGGGGDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK113hz52B2H9-Fc.LALA.YTE-73P1_scFv_LHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGSYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSA114參考抗IL-13 Ab #0 VL (CDR加底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYACWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL115參考抗IL-13 Ab #0 CDR-L1SGDKLGDKYAC116參考抗IL-13 Ab #0 LC (CL λ L2 wt) (VL加斜體；CDR加粗體)SYELTQPPSVSVSPGQTASITCSGDKLGDKYACWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS117抗IL-13 -LC.C34F (CL λ L1 wt) (CDR加底線；突變加粗體及底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAFWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS118抗IL-13 C34F-VLSYELTQPPSVSVSPGQTASITCSGDKLGDKYAFWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL119抗IL-13 C34F-CDR-L1SGDKLGDKYAF120抗IL-13 73P1-R2-L2-L135F (CL λ L2) (VL加斜體；CDR加底線；突變加粗體及底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKAAPSVTLFPPSSECLQANKATLVCFISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS121抗IL-13 73P1-R2-L2 (CL λ L2) (VL加斜體；CDR加底線；突變加粗體及底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKAAPSVTLFPPSSECLQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS122抗IL-13 73P1-L2.L135F (CL λ L2) (VL加斜體；CDR加底線；突變加粗體及底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKAAPSVTLFPPSSEELQANKATLVCFISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS123抗IL-13 73P1-HC-R2-LALA (VH加斜體；CDR加底線；突變加粗體及底線)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK124抗IL-13 73P1 VH-CH1 + 部分鉸鏈 (VH加底線及斜體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC125抗IL-13 73P1-Fc.LALA.YTEEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK126抗IL-13 73P1-IgG1 Fc LALA-LS-臼 – JT7/R2 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK127抗IL-13 73P1-IgG1 Fc IgG1 LALA-LS-杵 – JT7/R2 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK128抗IL-13 73P1-IgG1 Fc LALA-LS-臼 – JT11/R2 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK129抗IL-13 73P1-IgG1 Fc LALA-LS-杵 – JT11/R2 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK130抗IL-13 73P1-IgG1 Fc LALA-LS-杵 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK131抗IL-13 73P1-IgG1 Fc LALA-LS-臼 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK132抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-臼 – JT7/R2 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK133抗TSLP hz51B2-HC9-IgG1 Fc IgG1 LALA-LS-杵 – JT7/R2 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK134抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-臼 – JT11/R2 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK135抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-杵 – JT11/R2 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK136抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-杵 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK137抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-臼 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK138抗IL-13 73P1- IgG1 Fc LALA-LS-杵 – JT7 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK139抗IL-13 73P1-IgG1 Fc LALA-LS-臼 – JT7 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK140抗IL-13 73P1-IgG1 Fc LALA-LS-杵 – JT11 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK141抗IL-13 73P1-IgG1 Fc LALA-LS-臼 – JT11 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK               142抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-杵 – JT7 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK143抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-臼 – JT7 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK144抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-杵 – JT11 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK145抗TSLP hz51B2-HC9-IgG1 Fc LALA-LS-臼 – JT11 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK146抗IL-13 C34W-CDR-L1SGDKLGDKYAW147抗IL-13 C34W-VL (CDR加底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAWWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL148抗IL-13 C34M-CDR-L1SGDKLGDKYAM149抗IL-13 C34M-VL (CDR加底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAMWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL150抗IL-13 M34G-CDR-H1SYAGS151抗IL-13 M34G-VH (CDR加底線)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAGSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS152抗IL-13 M34F- CDR-H1SYAFS153抗IL-13 M34F-VH (CDR加底線)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAFSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS154抗IL-13 M34S-CDR-H1SYASS155抗IL-13 M34S-VH (CDR加底線)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYASSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS156抗IL-13 M34Y-CDR-H1SYAYS157抗IL-13 M34Y-VH (CDR加底線)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAYSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS158IgG1 CH1+ 部分鉸鏈域ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC159IgG1 CH1+ 部分鉸鏈域 – JT7ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC160IgG1 CH1 + 部分鉸鏈域 – JT11ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC161IgG1 CH1 + 部分鉸鏈域 – JT11/R2ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS162IgG1 CH1 + 部分鉸鏈域 – JT7/R2ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS163連接子GGSGG164連接子GSGSG165連接子GSGGG166連接子GGGSG167連接子GSSSG168連接子GGSGGS169連接子SGGGGS170連接子GRAGGGGAGGGG171連接子GRAGGG172連接子GGGGS173CL_λ_L135F (基於CL λ L2)GQPKAAPSVTLFPPSSEELQANKATLVCFISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS174CL λ_L135F/R2 (基於CL λ L2)GQPKAAPSVTLFPPSSECLQANKATLVCFISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS175抗TSLP hz51B2-LC3-R2.L135F (κ)(VL加斜體及底線；突變加粗體及底線)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIKRTVAAPSVFIFPPSDECLKSGTASVVCFLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGES176抗TSLP hz51B2-HC9-IgG1 Fc IgG1 LALA-LS-杵 – R2 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK177抗TSLP hz51B2-HC9-IgG1 Fc IgG1 LALA-LS-臼 – R2 (VH加底線及斜體；突變加底線及粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK178抗TSLP hz51B2-LC3-R2 κ (VL加斜體及底線；突變加粗體及底線)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIKRTVAAPSVFIFPPSDECLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGES179抗TSLP hz51B2-LC3-L135F (κ)(VL加斜體及底線；突變加粗體及底線)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCFLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC180CL_κ_L135FRTVAAPSVFIFPPSDEQLKSGTASVVCFLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC181CL κ_L135F/R2RTVAAPSVFIFPPSDECLKSGTASVVCFLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGES182抗IL-13  A33S/C34F-CDR-L1SGDKLGDKYSF183抗IL-13 C34L-CDR-L1SGDKLGDKYAL184抗IL-13 C34I- CDR-L1SGDKLGDKYAI185抗IL-13 C34A-CDR-L1SGDKLGDkYAA186抗IL-13 C34S-CDR-L1SGDKLGDKYAS187抗IL-13 C34H-CDR-L1SGDKLGDKYAH188抗TSLP HC9-stab. mut.QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMhWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS189抗TSLP LC3-stab. mut.DIQMTQSPSFLSASVGDRVTITCkASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGCGTKLEIK190抗TSLP LC4-stab. mut.DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK191抗IL-13 A33S/C34F VLSYELTQPPSVSVSPGQTASITCSGDKLGDKYSFWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL192抗IL-13 C34L VLSYELTQPPSVSVSPGQTASITCSGDKLGDKYALWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL193抗IL-13 C34I VLSYELTQPPSVSVSPGQTASITCSGDKLGDKYAIWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL194抗IL-13 C34A VLSYELTQPPSVSVSPGQTASITCSGDKLGDKYAAWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL195抗IL-13 C34S VLSYELTQPPSVSVSPGQTASITCSGDKLGDKYASWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL196抗IL-13 C34H VLSYELTQPPSVSVSPGQTASITCSGDKLGDKYAHWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL197抗IL-13 73P1.C34Y LC (CL λ L2 wt)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS198IgG1 LALA-LS-臼 – R2ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK199IgG1 LALA-LS-杵 – R2ASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK200抗IL-13 73P1-IgG1 Fc LALA-LS-臼 – R2 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK201抗IL-13 73P1-IgG1 Fc IgG1 LALA-LS-杵 – R2 (VH加底線及斜體；突變加底線及粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK202CL_λ_L135F (基於CL λ L1)GQPKANPTVTLFPPSSEELQANKATLVCFISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS203CL_λ_L135F/R2 (基於CL λ L1)GQPKANPTVTLFPPSSECLQANKATLVCFISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS204CL_λ_R2 (基於CL λ L1)GQPKANPTVTLFPPSSECLQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS205CL λ_R2 (基於CL λ L2)GQPKAAPSVTLFPPSSECLQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS206抗IL-13 73P1-R2 (CL λ L1) (VL加斜體；CDR加底線；突變加粗體及底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKANPTVTLFPPSSECLQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS207抗IL-13 73P1-L135F (CL λ L1) (VL加斜體；CDR加底線；突變加粗體及底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKANPTVTLFPPSSEELQANKATLVCFISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS208抗IL-13 73P1-L135F/R2 (CL λ L1) (VL加斜體；CDR加底線；突變加粗體及底線)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGQPKANPTVTLFPPSSECLQANKATLVCFISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS209抗IL-13 73P1-Fc.LALA.YTE-JT7EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP210抗IL-13 73P1-Fc.LALA.YTE-JT11EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP211抗IL-13 73P1-Fc.LALA.YTE-R2EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP212抗IL-13 73P1-Fc.LALA.YTE-JT7/R2EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTIPAVLQSSGLYSLLSLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP213抗IL-13 73P1-Fc.LALA.YTE-JT11/R2EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVCPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTVPAVLQSSGLYSLISLVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSSDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP214下鉸鏈部分DKTHTCPPCPA215上鉸鏈部分EPKSC216完整鉸鏈域#1EPKSCDKTHTCPPCPA217完整鉸鏈域#2EPKSSDKTHTCPPCPA218抗TSLP hz51B2-HC9LC3 scFv 無SS穩定作用 (連接子加粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIK219抗TSLP hz51B2-HC9LC4 scFv無SS穩定作用 (連接子加粗體)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGQGTKLEIK22073P1-Fc.LALA.LS1-hz51B2 H9L4 scFv HL-stab (連接子加粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGGGQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK22173P1-Fc.LALA.LS2-hz51B2 H9L4 scFv HL.V2 (stab) (連接子加粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGGGQVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK22273P1-Fc.LALA.LS3-hz51B2 H9L4 scFv HL (stab) (連接子加粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGGGQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSAGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK22373P1-Fc.LALA.LS4-hz51B2 H9L4 scFv HL.V2 (stab) (連接子加粗體)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGGGQVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSSAGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK224連接子AGGGGSGGGGSGGGGS22573P1-Fc.IgG1.LALA.LSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK226hz51B2 H9L4 scFv HL-3 (stab)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSAGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK227hz51B2 H9L4 scFv HL.V2-2 (stab)QVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK228hz51B2 VH9.V2-stabQVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSS229hz51B2 H9L4 scFv HL.V2-4 (stab)QVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSSAGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIKAll references cited in this application are incorporated herein by reference as if each reference were individually incorporated herein by reference. Although the description refers to specific embodiments, it will be apparent to those skilled in the art that the invention may be practiced with variations in these specific details. Therefore, the invention should not be construed as limited to the embodiments described herein.Sequence ListingSEQ ID NO:describesequence1 Anti-TSLP 51A1 VH (CDRs underlined) QVQLQQSGAELVRPGASVKLSCKASGYMFTDYAMH WVKQTPVHGLEWIGTIDPETGGTAYTQKFKG KATLTADISSSTAYVELRSLTSEDSAVYYCTREGSYGGYVDY WGQGTTLSVSS2 Anti-TSLP 51A1/51A4 CDR-H1 DYAMH3 Anti-TSLP 51A1/51A4 CDR-H2 TIDPETGGTAYTQKFKG4 Anti-TSLP 51A1/51A3/51A4/ 51A6 CDR-H3 EGSYGGYVDY5 Anti-TSLP 51A1VL (CDR underlined) DIVMTQSQKFMSTSVGDRVN ITCKASQNVHTAVA WYQQKPGQSPKTLIFLASNRHT GVPDRFTGSGSGTDFTLTISNVQSEDLADYFCLQLWNYPYT FGGETKLEIK6 Anti-TSLP 51A1/51A3/51A4/51A6 CDR-L1 KASQNVHTAVA7 Anti-TSLP 51A1/51A3/51A4/ 51A6 CDR-L2 LASNRHT8 Anti-TSLP 51A1/51A3/51A4/51A6 CDR-L3 LQLWNYPYT9 Anti-TSLP 51A4 VH (CDRs underlined) QVQLQQSGAELVRPGASVKLSCKASGYMFTDYAMH WVKQTPVHGLEWIGTIDPETGGTAYTQKFKG KATLTADISSNTAYVELRSLTSEDSAVYYCTREGSYGGYVDY WGQGTTLSVSS10 Anti-TSLP 51A4 VL (CDRs underlined) DIVMTQSQKFMSTSVGDRVS ITCKASQNVHTAVA WYQQKPGQSPKTLIFLASNRHT GVPDRFTGSGSGTDFTLTISNVQSEDLADYFCLQLWNYPYT FGGGTKL EIK11 Anti-TSLP 51A3/51A6 VH (CDRs underlined) QVQLQQSGAELVRPGASVKLSCKASGYTFTDYAMH WVKQTLVHGLEWVGTIDPETGGTAYTQKFQG KATLTADVSSSTAYVELRSLTSEDSAVYYCTREGSYGGYVDY WGQGTTLSVSS12 Anti-TSLP 51A3/51A6 CDR-H1 DYAMH13 Anti-TSLP 51A3/51A6 CDR-H2 TIDPETGGTAYTQKFQG14 Connector, where n is an integer at least 1 (GSGGS)n15 Anti-TSLP 51A6 VL Anti-TSLP 51A3 VL (CDRs underlined) DIVMTQSQKFMSTSE GDRVS ITCKASQNVHTAVA WYQQKPGQSPKTLIFLASNRHT GVPDRFTGSGSGTDFTLTISNVQSEDLADYFCLQLWNYPYT FGGGTKLEIK16 Connector, where n is an integer at least 1 (GGGS)n17 Connector, where n is an integer at least 1 (GGGGS)n18 connector GGSG19 Anti-TSLP 51B2 VH (CDRs underlined) QVQLLQPGTELVKPGASVKLSCKASGYTFTSYLMH WVKQRPGQGLEWIGEIDPSDSYTDYNQNFRG KATLTVDMSSSTAYMQLSSLTSEDSAVYYCALTGTINYFDY WGQGTTLTVSS20 anti-TSLP 51B2 CDR-H1 SYLMHtwenty one anti-TSLP 51B2 CDR-H2 EIDPSDSYTDYNQNFRGtwenty two anti-TSLP 51B2 CDR-H3 TGTINYFDYtwenty three Anti-TSLP 51B2 VL (CDRs underlined) DIVMTQSHKFMSTSVGDRVNITCKASQDVVTAVA WYQQKPGQSPKLLIYWASMRHT GVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSSFTFT FGSGTKLEIKtwenty four anti-TSLP 51B2 CDR-L1 KASQDVVTAVA25 anti-TSLP 51B2 CDR-L2 WASMRHT26 Anti-TSLP 51B2 CDR-L3 QQYSSFTFT27 Anti-TSLP 54A2 VH (CDRs underlined) QVQLRESGPGLVAPSQSLSITCTVSGFSLTSYGVN WVRQPPGKGLEWLGVIWDDGTTNYHSAFIS RLSITKDNSKSQVFLKLNSLQTDDTATYYCAKGSIYWYFDV WGAGTTVTVSS28 anti-TSLP 54A2 CDR-H1 SYGVN29 anti-TSLP 54A2 CDR-H2 VIWDDGTTNYHSAFIS30 anti-TSLP 54A2 CDR-H3 GSIYWYFDV31 Anti-TSLP 54A2 VL (CDRs underlined) DIVLTQSPASLAVSLGQRATISCKASQSVDYEGDSHMN WYQQKPGQSPKLLIYVASNLES GIPARFSGSGSGTDFTLNIHPVEEEDAATYYCQQSNEVPFT FGSGTKLEIK32 Anti-TSLP 54A2 CDR-L1 KASQSVDYEGDSHMN33 anti-TSLP 54A2 CDR-L2 VASNLES34 anti-TSLP 54A2 CDR-L3 QQSNEVPFT35 Anti-TSLP 54B3 VH (CDRs underlined) QVQLQQSGAELVRPGASVKLSCKASGYTFTDYEIH WVRQTPVHGLEWIGLIDPETGGTAHNQKFKA KATLTVDKSSTTAYMELRSLTSEDSAVYYCTREGQLGLSLDY WGQGTTLTVSS36 Anti-TSLP 54B3 CDR-H1 DYEIH37 Anti-TSLP 54B3 CDR-H2 LIDPETGGTAHNQKFKA38 anti-TSLP 54B3 CDR-H3 EGQLGLSLDY39 Anti-TSLP 54B3 VL (CDRs underlined) DIVMTQSHKFMSTSVGERVSITCKASQDVGTTVA WYQQKPGQSPKLLIYWTSTRHT GVPDRFTGSGSGTDFTLTISNVQSEDLADYFCQQYSTYPLT FGSGTKLEIQ40 Anti-TSLP 54B3 CDR-L1 KASQDVGTTVA41 Anti-TSLP 54B3 CDR-L2 WTSTRHT42 Anti-TSLP 54B3 CDR-L3 QQYSTYPLT43 Anti-TSLP 54C4 VH (CDRs underlined) EVQLQQSGPVLVKPGASVKMSCKASGYTFTDYYMN WVKQSHGKSLEWIGVINPYNADTKFNQRFKDK ATLTVDESSSTAYMELNSLTSEDSAVYYCARGDDYAEESFAY WGQGTLVTVSAA44 anti-TSLP 54C4 CDR-H1 DYYMN45 anti-TSLP 54C4 CDR-H2 VINPYNADTKFNQRFKD46 anti-TSLP 54C4 CDR-H3 GDDYAEESFAY47 Anti-TSLP 54C4 VL (CDRs underlined) QIVLTQSPSIMSASLGEEITLTCSASSSVSYMH WYQQKSGTSPKLLIYFTSNLAS GVPSRFSGSGSGTFYSLTISSVEAEDAADYYCHQWSSYPT FGGGTKLEIK48 anti-TSLP 54C4 CDR-L1 SASSSVSYMH49 anti-TSLP 54C4 CDR-L2 FTSNLAS50 anti-TSLP 54C4 CDR-L3 HQWSSYPT51 Anti-TSLP hz51B2 VL-1 (CDRs underlined) DIQLTQSPSFLSASVGDRVTITCKASQDVVTAVA WYQQKPGKAPKLLIYWASMRHT GVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFT FGQGTKLEIK52 Anti-TSLP hz51B2 VL-2 (CDRs underlined) DIQLTQSPSFLSASVGDRVTITCKASQDVVTAVA WYQQKPGKAPKLLIYWASMRHT GVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFT FGQGTKLEIK53 Anti-TSLP hz51B2 VL-3 (CDRs underlined) DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVA WYQQKPGKAPKLLIYWASMRHT GVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFT FGQGTKLEIK54 Anti-TSLP hz51B2 VL-4 (CDRs underlined) DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVA WYQQKPGKAPKLLIYWASMRHT GVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFT FGQGTKLEIK55 Anti-TSLP hz51B2 VH-1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQG LEWMGEIDPSDSYTDYNQNFRG RVTMTRDTSTSTVYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS56 Anti-TSLP hz51B2 VH-2 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQG LEWMGEIDPSDSYTDYNQNFRG RVTMTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS57 Anti-TSLP hz51B2 VH-3 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQG LEWIGEIDPSDSYTDYNQNFRG RVTMTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS58 Anti-TSLP hz51B2 VH-4 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQG LEWMGEIDPSDSYTDYNQNFRG RVTLTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS59 Anti-TSLP hz51B2 VH-5 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQG LEWIGEIDPSDSYTDYNQNFRG RVTLTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS60 Anti-TSLP hz51B2 VH-6 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQG LEWMGEIDPSDSYTDYNQNFRG RATMTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS61 Anti-TSLP hz51B2 VH-7 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQG LEWIGEIDPSDSYTDYNQNFRG RATMTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS62 Anti-TSLP hz51B2 VH-8 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQG LEWIGEIDPSDSYTDYNQNFRG RATLTVDMSTSTVYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS63 Anti-TSLP hz51B2 VH-9 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQR LEWMGEIDPSDSYTDYNQNFRG RVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS64 Anti-TSLP hz51B2 VH-10 QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMH WVRQAPGQR LEWIGEIDPSDSYTDYNQNFRG RVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDY WGQGTLVTVSS65 Anti-IL-13 73P1 VH (CDRs underlined) EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMS WVRQAPGKGLEWVSAFSGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDV WGQGTTVTVSS66 Anti-IL-13 73P1 CDR-H1 SYAMS67 Anti-IL-13 73P1 CDR-H2 AFSGSGGSTYYADSVKG68 Anti-IL-13 73P1 CDR-H3 DGLGPYFYNYGMDV69 Anti-IL-13 73P1 VL (CDRs underlined) SYELTQPPSVSVSPGQTASITCSGDKLGDKYAY WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL70 Anti-IL-13 73P1 CDR-L1 SGDKLGDKYAY71 Anti-IL-13 73P1 CDR-L2 HDSKRPS72 Anti-IL-13 73P1 CDR-L3 QAWDSSTYV73 CL κ wt RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC74 CL λ L2 wt GQPKAA PS VTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADS SPVKAGVETTT PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS75 CL λ L1 wt GQPKAN PT VTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADG SPVKAGVETTK PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS76 Human IgG1 Fc DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK77 IgG1 Fc-LALA DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK78 IgG1 Fc-LALA-LS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK79 IgG1 Fc-LALA-YTE DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK80 IgG1 Fc-LALA-LS-mortar DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK81 IgG1 Fc-LALA-LS-Pestle DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK82 Human IgG1 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK*83 IgG1 LALA ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK84 IgG1 LALA-LS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAP IEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK85 IgG1 LALA-LS-Pestle ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK86 IgG1 LALA-LS-mortar ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK87 IgG1 LALA-LS-JT7 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK88 IgG1 LALA-LS-Pestle – JT7 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK89 IgG1 LALA-LS-JT11 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK90 IgG1 LALA-LS-Pestle – JT11 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK91 IgG1 LALA-LS-JT7/R2 ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK92 IgG1 LALA-LS-Pestle – JT7/R2 ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK93 IgG1 LALA-LS-JT11/R2 ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK94 IgG1 LALA-LS-Pestle – JT11/R2 ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK95 IgG1 Fc-LALA-HA DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK96 IgG1 Fc-LALA-knob DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK97 connector PLGLAG98 Connector 1 GGGG99 Connector 2 GGGGSGGGGSGGGGS101 Anti-IL-13 73P1-HC-LALA (VH italicized and underlined)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK102 Anti-IL-13 73P1-LC.C34Y (CL λ L1 wt) (VL italicized and underlined)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVL GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS103 Anti-TSLP hz51B2-LC3 (CL κ wt) (VL italicized and underlined)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC104 Anti-TSLP hz51B2-HC9 (IgG1 LALA) (VH italicized and underlined)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK105 Anti-TSLP hz51B2-HC9 (IgG1 LALA-YTE) (VH italicized and underlined)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK106 Anti-TSLP hz51B2-HC9LC3 scFv-stab (linker in bold) QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGCGTKLEIK107 Anti-TSLP hz51B2-HC9LC4 scFv-stab (linker in bold) QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK108 Anti-IL-13 73P1 scFv-LH (linker in bold)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAY WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVLGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMS WVRQAPGKGLEWVSAFSGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDV WGQGTTVTVSS109 Anti-TSLP hz51B2-HC9 VH-CH1 (VH italicized and underlined)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC110 (hz51B2-H9-CH1)-(73P1_scFv-LH)-Fc.LALA.YTE (hz51B2-H9-VH italicized and underlined; linker in bold; 73P1-VH underlined; 73P1-VL italicized)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCGGGGSYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSGGGG DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK111 73P1-Fc.LALA.YTE-hz51B2 H9L4 scFv HL-stab (linker in bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK112 73P1HC-hz51B2.H9L4 scFv HL-stab-Fc.LALA.YTEEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSSCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIKGGGG DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK113 hz52B2H9-Fc.LALA.YTE-73P1_scFv_LHQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGSYELTQPPSVSVSPGQTASITCSGDKLGDKYAYWFQQKPGQSPVLVIYHDSKRPSGIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYVFGTGTKVTVLGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS A114 Reference anti-IL-13 Ab #0 VL (CDRs underlined) SYELTQPPSVSVSPGQTASITCSGDKLGDKYAC WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL115 Reference anti-IL-13 Ab #0 CDR-L1 SGDKLGDKYAC116 Reference anti-IL-13 Ab #0 LC (CLλL2 wt) (VL in italics; CDR in bold)T PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS117 Anti-IL-13 -LC.C34F (CL λ L1 wt) (CDRs underlined; mutations bold and underlined)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAF WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS118 Anti-IL-13 C34F-VL SYELTQPPSVSVSPGQTASITCSGDKLGDKYAF WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL119 Anti-IL-13 C34F-CDR-L1 SGDKLGDKYAF120 Anti-IL-13 73P1-R2-L2-L135F (CL λ L2) (VL italicized; CDRs underlined; mutations bolded and underlined)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAY WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL GQPKAAPSVTLFPPSSEC LQANKATLVCF ISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTES S121 Anti-IL-13 73P1-R2-L2 (CL λ L2) (VL italicized; CDRs underlined; mutations bolded and underlined)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAY WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL GQPKAAPSVTLFPPSSEC LQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTES S122 Anti-IL-13 73P1-L2.L135F (CL λ L2) (VL italicized; CDRs underlined; mutations bolded and underlined)F ISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS123 Anti-IL-13 73P1-HC-R2-LALA (VH in italics; CDRs underlined; mutations in bold and underlined)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMS WVRQAPGKGLEWVSAFSGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDV WGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKG QPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK124 Anti-IL-13 73P1 VH-CH1 + partial hinge (VH underlined and italicized)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC125 Anti-IL-13 73P1-Fc.LALA.YTEEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTL PPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK126 Anti-IL-13 73P1-IgG1 Fc LALA-LS-hole – JT7/R2 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK127 Anti-IL-13 73P1-IgG1 Fc IgG1 LALA-LS-knob – JT7/R2 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK128 Anti-IL-13 73P1-IgG1 Fc LALA-LS-hole – JT11/R2 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK129 Anti-IL-13 73P1-IgG1 Fc LALA-LS-knob – JT11/R2 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK130 Anti-IL-13 73P1-IgG1 Fc LALA-LS-knob (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK131 Anti-IL-13 73P1-IgG1 Fc LALA-LS-hole (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK132 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-Area – JT7/R2 (VH underlined and italicized; mutations underlined and bold)I VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK133 Anti-TSLP hz51B2-HC9-IgG1 Fc IgG1 LALA-LS-knob – JT7/R2 (VH underlined and italicized; mutations underlined and bold)IW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK134 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-Area – JT11/R2 (VH underlined and italicized; mutations underlined and bold)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK135 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-knob – JT11/R2 (VH underlined and italicized; mutations underlined and bold)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK136 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-knob (VH underlined and italicized; mutations underlined and bold)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK137 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-hole (VH underlined and italicized; mutations underlined and bold)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK138 Anti-IL-13 73P1- IgG1 Fc LALA-LS-knob – JT7 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK139 Anti-IL-13 73P1-IgG1 Fc LALA-LS-hole – JT7 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK140 Anti-IL-13 73P1-IgG1 Fc LALA-LS-knob – JT11 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK141 Anti-IL-13 73P1-IgG1 Fc LALA-LS-hole – JT11 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK142 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-knob – JT7 (VH underlined and italicized; mutations underlined and bold)IPAVLQSSGLYSLL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK143 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-Area – JT7 (VH underlined and italicized; mutations underlined and bold)IPAVLQSSGLYSLL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK144 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-knob – JT11 (VH underlined and italicized; mutations underlined and bold)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK145 Anti-TSLP hz51B2-HC9-IgG1 Fc LALA-LS-Area – JT11 (VH underlined and italicized; mutations underlined and bold)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQR LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK146 Anti-IL-13 C34W-CDR-L1 SGDKLGDKYAW147 Anti-IL-13 C34W-VL (CDR underlined) SYELTQPPSVSVSPGQTASITCSGDKLGDKYAW WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL148 Anti-IL-13 C34M-CDR-L1 SGDKLGDKYAM149 Anti-IL-13 C34M-VL (CDR underlined) SYELTQPPSVSVSPGQTASITCSGDKLGDKYAM WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL150 Anti-IL-13 M34G-CDR-H1 SYAGS151 Anti-IL-13 M34G-VH (CDRs underlined) EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAGS WVRQAPGKGLEWVSAFSGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDV WGQGTTVTVSS152 Anti-IL-13 M34F- CDR-H1 SYAFS153 Anti-IL-13 M34F-VH (CDRs underlined) EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAFS WVRQAPGKGLEWVSAFSGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDV WGQGTTVTVSS154 Anti-IL-13 M34S-CDR-H1 SYASS155 Anti-IL-13 M34S-VH (CDRs underlined) EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYASS WVRQAPGKGLEWVSAFSGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDV WGQGTTVTVSS156 Anti-IL-13 M34Y-CDR-H1 SYAYS157 Anti-IL-13 M34Y-VH (CDRs underlined) EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAYS WVRQAPGKGLEWVSAFSGSGGSTYYADSVKG RFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDV WGQGTTVTVSS158 IgG1 CH1+ partial hinge domain ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC159 IgG1 CH1+ partial hinge domain – JT7 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC160 IgG1 CH1 + partial hinge domain – JT11 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC161 IgG1 CH1 + partial hinge domain – JT11/R2 ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS162 IgG1 CH1 + partial hinge domain – JT7/R2 ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS163 connector GGSGG164 connector GSGSG165 connector GSGGG166 connector GGGSG167 connector GSSSG168 connector GGSGGS169 connector SGGGGS170 connector GRAGGGGAGGGG171 connector GRAGGG172 connector GGGGS173 CL_λ_L135F (based on CL λ L2) GQPKAAPSVTLFPPSSEELQANKATLVCF ISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS174 CL λ_L135F/R2 (Based on CL λ L2) GQPKAAPSVTLFPPSSEC LQANKATLVCF ISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTES175 Anti-TSLP hz51B2-LC3-R2.L135F (κ) (VL in italics and underline; mutation in bold and underline)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIK RTVAAPSVFIFPPSDEC LKSGTASVVCF LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGES176 Anti-TSLP hz51B2-HC9-IgG1 Fc IgG1 LALA-LS-knob – R2 (VH underlined and italicized; mutations underlined and bold)C PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK177 Anti-TSLP hz51B2-HC9-IgG1 Fc IgG1 LALA-LS-hole – R2 (VH underlined and italicized; mutations underlined and bold)C PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK178 Anti-TSLP hz51B2-LC3-R2 κ (VL in italics and underline; mutation in bold and underline)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIK RTVAAPSVFIFPPSDEC LKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGES179 Anti-TSLP hz51B2-LC3-L135F (κ) (VL in italics and underline; mutation in bold and underline)DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCF LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC180 CL_κ_L135F RTVAAPSVFIFPPSDEQLKSGTASVVCF LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC181 CL κ_L135F/R2 RTVAAPSVFIFPPSDEC LKSGTASVVCF LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGES182 Anti-IL-13 A33S/C34F-CDR-L1 SGDKLGDKYSF183 Anti-IL-13 C34L-CDR-L1 SGDKLGDKYAL184 Anti-IL-13 C34I- CDR-L1 SGDKLGDKYAI185 Anti-IL-13 C34A-CDR-L1 SGDKLGDkYAA186 Anti-IL-13 C34S-CDR-L1 SGDKLGDKYAS187 Anti-IL-13 C34H-CDR-L1 SGDKLGDKYAH188 Anti-TSLP HC9-stab. mut. QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMhWVRQAPGQC LEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSS189 Anti-TSLP LC3-stab. mut. DIQMTQSPSFLSASVGDRVTITCkASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGC GTKLEIK190 Anti-TSLP LC4-stab. mut. DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGC GTKLEIK191 Anti-IL-13 A33S/C34F VL SYELTQPPSVSVSPGQTASITCSGDKLGDKYSF WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL192 Anti-IL-13 C34L VL SYELTQPPSVSVSPGQTASITCSGDKLGDKYAL WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL193 Anti-IL-13 C34I VL SYELTQPPSVSVSPGQTASITCSGDKLGDKYAI WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL194 Anti-IL-13 C34A VL SYELTQPPSVSVSPGQTASITCSGDKLGDKYAA WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL195 Anti-IL-13 C34S VL SYELTQPPSVSVSPGQTASITCSGDKLGDKYAS WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL196 Anti-IL-13 C34H VL SYELTQPPSVSVSPGQTASITCSGDKLGDKYAH WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL197 Anti-IL-13 73P1.C34Y LC (CLλ L2 wt)T PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS198 IgG1 LALA-LS-R2 ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK199 IgG1 LALA-LS-Pestle – R2 ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK200 Anti-IL-13 73P1-IgG1 Fc LALA-LS-hole – R2 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLS CA VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLV SKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK201 Anti-IL-13 73P1-IgG1 Fc IgG1 LALA-LS-knob – R2 (VH underlined and italicized; mutations underlined and bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLW CLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVL HEALHS HYTQKSLSLSPGK202 CL_λ_L135F (based on CL λ L1) GQPKAN PT VTLFPPSSEELQANKATLVCF ISDFYPGAVTVAWKADG SPVKAGVETTK PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS203 CL_λ_L135F/R2 (based on CL λ L1) GQPKAN PT VTLFPPSSEC LQANKATLVCF ISDFYPGAVTVAWKADG SPVKAGVETTK PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTES S204 CL_λ_R2 (based on CL λ L1) GQPKAN PT VTLFPPSSEC LQANKATLVCLISDFYPGAVTVAWKADG SPVKAGVETTK PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTES S205 CL λ_R2 (based on CL λ L2) GQPKAAPSVTLFPPSSEC LQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTES S206 Anti-IL-13 73P1-R2 (CL λ L1) (VL italicized; CDRs underlined; mutations bolded and underlined)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAY WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL GQPKAN PT VTLFPPSSEC LQANKATLVCLISDFYPGAVTVAWKADG SPVKAGVETTK PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTES S207 Anti-IL-13 73P1-L135F (CL λ L1) (VL italicized; CDRs underlined; mutations bolded and underlined)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAY WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL GQPKAN PT VTLFPPSSEELQANKATLVCF ISDFYPGAVTVAWKADG SPVKAGVETTK PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS208 Anti-IL-13 73P1-L135F/R2 (CL λ L1) (VL italicized; CDRs underlined; mutations bolded and underlined)SYELTQPPSVSVSPGQTASITCSGDKLGDKYAY WFQQKPGQSPVLVIYHDSKRPS GIPERFSGSNSGDTATLTISGTQAMDEADYYCQAWDSSTYV FGTGTKVTVL GQPKAN PT VTLFPPSSEC LQANKATLVCF ISDFYPGAVTVAWKADG SPVKAGVETTK PSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTES S209 Anti-IL-13 73P1-Fc.LALA.YTE-JT7EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP210 Anti-IL-13 73P1-Fc.LALA.YTE-JT11EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP211 Anti-IL-13 73P1-Fc.LALA.YTE-R2EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP212 Anti-IL-13 73P1-Fc.LALA.YTE-JT7/R2EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTI PAVLQSSGLYSLL SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP213 Anti-IL-13 73P1-Fc.LALA.YTE-JT11/R2EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVC PLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTV PAVLQSSGLYSLI SL VTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSS DKTHTCPPCPAPEAA GGPSVFLFPPKPKDTLY IT RE PEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSP214 Lower hinge part DKTHTCPPCPA215 Upper hinge part EPKSC216 Complete Hinge #1 EPKSCDKTHTCPPCPA217 Complete Hinge #2 EPKSSDKTHTCPPCPA218 Anti-TSLP hz51B2-HC9LC3 scFv has no SS stabilization effect (linker in bold) QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCQQYSSFTFTFGQGTKLEIK219 Anti-TSLP hz51B2-HC9LC4 scFv has no SS stabilization effect (linker in bold) QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQRLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGS DIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGQGTKLEIK220 73P1-Fc.LALA.LS1-hz51B2 H9L4 scFv HL-stab (linker in bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGGGQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK221 73P1-Fc.LALA.LS2-hz51B2 H9L4 scFv HL.V2 (stab) (linker in bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGGGQVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK222 73P1-Fc.LALA.LS3-hz51B2 H9L4 scFv HL (stab) (linker in bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGGGQVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSAGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK223 73P1-Fc.LALA.LS4-hz51B2 H9L4 scFv HL.V2 (stab) (linker in bold)EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGGGQVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSSAGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK224 connectorAGGGGSGGGGSGGGGS225 73P1-Fc.IgG1.LALA.LSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAFSGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCVQDGLGPYFYNYGMDVWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGV EVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVLHEALHSHYTQKSLSLSPGK226 hz51B2 H9L4 scFv HL-3 (stab)QVQLVQSGAEVKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTAVYYCALTGTINYFDYWGQGTLVTVSSAGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK227 hz51B2 H9L4 scFv HL.V2-2 (stab)QVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSSGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK228 hz51B2 VH9.V2-stabQVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSS229 hz51B2 H9L4 scFv HL.V2-4 (stab)QVQLVQSGAESKKPGASVKVSCKASGYTFTSYLMHWVRQAPGQCLEWMGEIDPSDSYTDYNQNFRGRVTITVDMSASTAYMELSSLRSEDTATYYCALTGTINYFDYWGQGTTVTVSSAGGGGSGGGGSGGGGSDIQMTQSPSFLSASVGDRVTITCKASQDVVTAVAWYQQKPGKAPKLLIYWASMRHTGVPSRFSGSGSGTEFTLTISSLQPEDFATYFCQQYSSFTFTFGCGTKLEIK

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圖1顯示了當以示例性抗TSLP抗體處理時，經由表現在STAT5-HEK293報導細胞上的TSLP受體複合物的TSLP傳訊的抑制百分比。Figure1 shows the percent inhibition of TSLP signaling through TSLP receptor complexes expressed on STAT5-HEK293 reporter cells when treated with exemplary anti-TSLP antibodies.

圖2顯示了在ELISA競爭實驗中51B2及US-FDA批准的參考抗TSLP Ab結合至TSLP上重合但不同的表位。Ab，抗體；OD，光學密度。Figure2 shows that 51B2 and a US-FDA-approved reference anti-TSLP Ab bind to overlapping but distinct epitopes on TSLP in a competitive ELISA assay. Ab, antibody; OD, optical density.

圖3A-3B顯示了與US-FDA批准的參考抗TSLP Ab相比，51B2完全抑制TSLP誘導的PBMC (圖3A)或分離的樹突細胞(圖3B)的CCL17分泌。Figures3A-3B show that 51B2 completely inhibited TSLP-induced CCL17 secretion by PBMCs (Figure3A ) or isolated dendritic cells (Figure3B ) compared to a US-FDA-approved reference anti-TSLP Ab.

圖4顯示了在使用TSLP受體複合體轉染的HEK293報導細胞進行的人類及食蟹獼猴 (「cyno」) TSLP抑制試驗中，與親代Ch51B2抗體相比，各個產生的人源化51B2抗體所得的IC50(nM)值及IC90(nM)值。由方框標記的人源化抗TSLP抗體表明被選來進行進一步分析的抗TSLP抗體。Figure4 shows the IC50 (nM) and IC90 (nM) values obtained for each humanized 51B2 antibody compared to the parental Ch51B2 antibody in a human and cynomolgus macaque ("cyno") TSLP inhibition assay usingHEK293 reporter cells transfected with the TSLP receptor complex. The humanized anti-TSLP antibodies marked by the box indicate the anti-TSLP antibodies selectedfor further analysis.

圖5A顯示了當以所選的人源化抗TSLP抗體(hz51B2 L2H2、hz51B2 L2H9及hz51B2 L3H9)處理時，經由表現在STAT5-HEK293報導細胞上的TSLP受體複合物的TSLP傳訊的抑制百分比。Ch51B2親代嵌合抗體作為對照。Figure5A shows the percent inhibition of TSLP signaling via TSLP receptor complexes expressed on STAT5-HEK293 reporter cells when treated with selected humanized anti-TSLP antibodies (hz51B2 L2H2, hz51B2 L2H9, and hz51B2 L3H9). The parental chimeric Ch51B2 antibody served as a control.

圖5B顯示了hz51B2 L3H9在溶液中結合至人類TSLP的平衡結合分析。Figure5B shows the equilibrium binding analysis of hz51B2 L3H9 binding to human TSLP in solution.

圖6A-6B顯示了在HEK-Blue 293 IL-13報導試驗中，抗IL-13抗體73P1增強對食蟹獼猴 (「cyno」) IL-13的中和活性，同時維持對人類IL-13抑制的中和活性，其中分泌的胚胎鹼性磷酸酯酶(SEAP)係於IL-13傳訊活化後產生。圖6A顯示了使用IL-13報導試驗，在多個濃度下，參考抗IL-13抗體#0及抗IL-13抗體73P1針對人類IL-13的中和活性，並提供兩種抗IL-13抗體的IC50。圖6B顯示了使用IL-13報導試驗，在多個濃度下，參考抗IL-13抗體#0及抗IL-13抗體73P1針對食蟹獼猴IL-13的中和活性，並進一步提供兩種抗IL-13抗體的IC50。Conc.，濃度；OD，光學密度。Figures6A-6B show that anti-IL-13 antibody 73P1 enhances the neutralization activity of cynomolgus macaque ("cyno") IL-13 in a HEK-Blue 293 IL-13 reporter assay, while maintaining its inhibitory neutralization activity against human IL-13. Secreted embryonic alkaline phosphatase (SEAP) is produced upon activation of IL-13 signaling.Figure6A shows the neutralization activity of reference anti-IL-13 antibody #0 and anti-IL-13 antibody 73P1 against human IL-13 at various concentrations using the IL-13 reporter assay, and provides theIC50 values for both anti-IL-13 antibodies.Figure6B shows the neutralizing activity of reference anti-IL-13 antibody #0 and anti-IL-13 antibody 73P1 against cynomolgus macaque IL-13 at various concentrations using an IL-13 reporter assay. TheIC50 values for the two anti-IL-13 antibodies are also provided. Conc., concentration; OD, optical density.

圖7A-7B顯示了使用HEK-Blue 293 IL-13報導試驗，抗IL-13抗體73P1及兩種US-FDA或EMA批准的作為陽性對照的抗IL-13抗體參考抗IL-13抗體#1及參考抗IL-13抗體#2 (「參考Ab. #1」及「參考Ab. #2」)的中和活性比較，其中使用野生型(「WT」) IL-13或IL-13 R110Q (IL-13的疾病相關變體)作為刺激物，在IL-13傳訊活化後產生分泌的胚胎鹼性磷酸酯酶(SEAP)。圖7A顯示了抗IL-13抗體73P1及參考抗IL-13 Ab. #1及#2 針對野生型人類IL-13的中和活性。圖7B顯示了抗IL-13抗體73P1及參考抗IL-13 Ab. #1及#2針對疾病相關人類IL-13變體(R110Q)的中和活性，其表現出IL-13活性增加。提供IC50值。Figures7A-7B show a comparison of the neutralizing activity of anti-IL-13 antibody 73P1 and two US-FDA- or EMA-approved anti-IL-13 antibodies, Reference Anti-IL-13 Antibody #1 and Reference Anti-IL-13 Antibody #2 ("Reference Ab. #1" and "Reference Ab. #2"), as positive controls, using a HEK-Blue 293 IL-13 reporter assay, using wild-type ("WT") IL-13 or IL-13 R110Q (a disease-associated variant of IL-13) as stimuli to produce secreted embryonic alkaline phosphatase (SEAP) upon activation of IL-13 signaling.Figure7A shows the neutralizing activity of anti-IL-13 antibody 73P1 and Reference Anti-IL-13 Abs. #1 and #2 against wild-type human IL-13.Figure7B shows the neutralizing activity of anti-IL-13 antibody 73P1 and reference anti-IL-13 Abs #1 and #2 against a disease-associated human IL-13 variant (R110Q), which exhibits increased IL-13 activity.IC50 values are provided.

圖8A-8D顯示了一組基於scFv融合及插入的示例性雙特異性抗TSLP×IL-13抗體構築體的示意圖。Figures8A-8D show schematic diagrams of a set of exemplary bispecific anti-TSLP x IL-13 antibody constructs based on scFv fusion and insertion.

圖9A-9B證實了圖8A-8D所示之TSLP×IL13雙特異性抗體針對TSLP及IL-13的抑制活性。圖9A顯示了當以雙特異性抗體之一者處理時，經由表現在STAT5-HEK293報導細胞上的TSLPR/IL-7Rα受體複合物的TSLP傳訊的抑制百分比。由報導細胞產生的螢光素酶作為TSLP結合活性的讀數。圖9B顯示了當以雙特異性抗體之一者處理時，經由表現在HEK293報導細胞上的IL-13Rα1/IL-4Rα受體複合物的IL-13傳訊的抑制百分比。由報導細胞產生及分泌的SEAP作為IL-13結合活性的讀數。根據抗體劑量依賴性結合抑制曲線，計算每種抗體針對抗原的IC90值。使用GraphPad分析及呈現數據。Ab，抗體；Conc.，濃度。Figures9A-9B demonstrate the inhibitory activity of the TSLP×IL13 bispecific antibodies shown inFigures8A-8D against TSLP and IL-13.Figure9A shows the percent inhibition of TSLP signaling via the TSLPR/IL-7Rα receptor complex expressed on STAT5-HEK293 reporter cells when treated with one of the bispecific antibodies. Luciferase produced by the reporter cells served as a readout for TSLP binding activity.Figure9B shows the percent inhibition of IL-13 signaling via the IL-13Rα1/IL-4Rα receptor complex expressed on HEK293 reporter cells when treated with one of the bispecific antibodies. SEAP produced and secreted by the reporter cells served as a readout for IL-13 binding activity. Based on the dose-dependent binding inhibition curves, theIC90 value of each antibody against the antigen was calculated. Data were analyzed and presented using GraphPad. Ab, antibody; Conc., concentration.

圖10顯示了圖8A-8D所示之TSLP×IL-13雙特異性抗體的體內穩定性，其係藉由將每種雙特異性抗體的抗TSLP抗原結合部分及抗IL-13抗原結合部分靜脈注射至C57Bl/6小鼠後隨時間變化的體內抗體濃度來呈現。針對每種抗體的每個稀釋測試兩隻動物(n=2/組)。Figure10 shows the in vivo stability of the TSLP×IL-13 bispecific antibodies shown inFigures8A-8D . This is demonstrated by measuring the time-dependent in vivo antibody concentrations following intravenous injection of the anti-TSLP and anti-IL-13 antigen-binding portions of each bispecific antibody into C57Bl/6 mice. Two animals (n = 2/group) were tested for each dilution of each antibody.

圖11A-11D顯示了一組基於Fab融合(分別為形式PX128-I1至PX128-I4)的示例性雙特異性TSLP×IL-13抗體的示意圖。Figures11A-11D show schematics of a panel of exemplary bispecific TSLP x IL-13 antibodies based on Fab fusions (formats PX128-I1 to PX128-I4, respectively).

圖12A-12B證實了圖11A-11D所示之形式PX128-I1至PX128-I4及圖13所示之PX128-R2的TSLP×IL-13雙特異性抗體針對TSLP (圖12A)及IL-13 (圖12B)的抑制活性。圖12A顯示了當以雙特異性抗體之一者處理時，經由表現在STAT5-HEK-293報導細胞上的TSLP受體複合物的TSLP傳訊的抑制百分比。圖12B顯示了當以雙特異性抗體之一者處理時，經由表現在HEK-293報導細胞上的IL-13受體複合體的IL-13傳訊的抑制百分比。抗IL-13參考抗體#2、#3及#4作為陽性對照。根據抗體劑量依賴性結合抑制曲線，計算出每種抗體針對抗原的IC90值。使用GraphPad分析及呈現數據。Ab，抗體；Conc.，濃度。Figures12A-12B demonstrate the inhibitory activity of TSLP×IL-13 bispecific antibodies, formats PX128-I1 to PX128-I4 shown inFigures11A- 11D, and PX128-R2 shown inFigure13 , against TSLP (Figure12A ) and IL-13 (Figure12B ).Figure12A shows the percent inhibition of TSLP signaling via the TSLP receptor complex expressed on STAT5-HEK-293 reporter cells when treated with one of the bispecific antibodies.Figure12B shows the percent inhibition of IL-13 signaling via the IL-13 receptor complex expressed on HEK-293 reporter cells when treated with one of the bispecific antibodies. Anti-IL-13 reference antibodies #2, #3, and #4 served as positive controls. Based on the dose-dependent binding inhibition curves, theIC90 value for each antibody against the antigen was calculated. Data were analyzed and presented using GraphPad. Ab, antibody; Conc., concentration.

圖13提供了PX128-R2異二聚體雙特異性抗體的示意圖，其中第一臂包括抗TSLP Fab片段，且第二臂包括抗IL-13 Fab片段。Fc域包括杵臼(knob-in-hole)突變。抗IL-13 Fab片段的CH1域及CL域亦包括R2突變，其包括抗IL-13臂之重鏈中的突變F126C及C220S以及輕鏈中的突變 E124C及C214S (所有EU編號)。Figure13 provides a schematic diagram of the PX128-R2 heterodimeric bispecific antibody, wherein the first arm comprises an anti-TSLP Fab fragment and the second arm comprises an anti-IL-13 Fab fragment. The Fc domain comprises knob-in-hole mutations. The CH1 and CL domains of the anti-IL-13 Fab fragment also comprise R2 mutations, including mutations F126C and C220S in the heavy chain of the anti-IL-13 arm and mutations E124C and C214S in the light chain (all EU numberings).

圖14提供了PX128-JT異二聚體雙特異性抗體的示意圖，其中第一臂包括抗TSLP Fab片段，且第二臂包括抗IL-13 Fab片段。Fc域包括杵臼突變。抗IL-13 Fab片段的CH1域及CL域亦包括JT11或JT7突變。JT11突變包括抗IL-13臂之HC中的F170V、S183I及V185L及LC中的L135F (所有EU編號)。JT7突變包括抗IL-13臂之HC中的F170I、S183L及V185L以及LC中的L135F (所有EU編號)。Figure14 provides a schematic diagram of the PX128-JT heterodimeric bispecific antibody, wherein the first arm comprises an anti-TSLP Fab fragment and the second arm comprises an anti-IL-13 Fab fragment. The Fc domain comprises a knob-to-hole mutation. The CH1 and CL domains of the anti-IL-13 Fab fragment also comprise JT11 or JT7 mutations. The JT11 mutations comprise F170V, S183I, and V185L in the HC of the anti-IL-13 arm and L135F in the LC (all EU numbering). The JT7 mutations comprise F170I, S183L, and V185L in the HC of the anti-IL-13 arm and L135F in the LC (all EU numbering).

圖15提供了PX128-JT/R2異二聚體雙特異性抗體的示意圖，其中第一臂包括抗TSLP Fab片段，且第二臂包括抗IL-13 Fab片段。Fc域包括杵臼突變。抗IL-13臂之重鏈及輕鏈亦包括R2突變及JT11或JT7突變(HC：JT11/R2 – HC：F126C、F170V、S183I、V185L及C220S或JT7/R2 – HC：F126C、F170I、S183L、V185L及C220S；LC：E124C、L135F及C214S)。Figure15 provides a schematic diagram of the PX128-JT/R2 heterodimeric bispecific antibody, in which the first arm comprises an anti-TSLP Fab fragment and the second arm comprises an anti-IL-13 Fab fragment. The Fc domain includes a knob-to-hole mutation. The heavy and light chains of the anti-IL-13 arm also include R2 mutations and either JT11 or JT7 mutations (HC: JT11/R2 – HC: F126C, F170V, S183I, V185L, and C220S or JT7/R2 – HC: F126C, F170I, S183L, V185L, and C220S; LC: E124C, L135F, and C214S).

圖16顯示了(圖14所示之) PX128-JT7及PX128-JT11異二聚體雙特異性抗體的體內穩定性，其係藉由將抗TSLP抗原結合部分及抗IL-13抗原結合部分靜脈注射至C57Bl/6小鼠後隨時間變化的體內抗體濃度來呈現。針對每種抗體的每個濃度測試兩隻動物(n=2/組)。Figure16 shows the in vivo stability of the PX128-JT7 and PX128-JT11 heterodimeric bispecific antibodies (shown inFigure14 ), as measured by time-dependent in vivo antibody concentrations following intravenous injection of the anti-TSLP and anti-IL-13 antigen-binding moieties into C57Bl/6 mice. Two animals (n = 2/group) were tested at each concentration of each antibody.

圖17A-17B證實了圖13-15所示之異二聚體雙特異性抗體針對TSLP (圖17A)及IL-13 (圖17B)的抑制活性。圖17A顯示了當以異二聚體雙特異性抗體之一者處理時，經由表現在STAT5-HEK-293報導細胞上的TSLP受體複合物的TSLP傳訊的抑制百分比。圖17B顯示了當以異二聚體雙特異性抗體之一者處理時，經由表現在HEK-293-SEAP 報導細胞上的IL-13受體複合體的IL-13傳訊的抑制百分比。根據抗體劑量依賴性結合抑制曲線，計算出每種抗體針對抗原的IC90值。使用GraphPad分析及呈現數據。Ab，抗體；Conc.，濃度。Figures17A-17B demonstrate the inhibitory activity of the heterodimeric bispecific antibodies shown inFigures13-15 against TSLP (Figure17A ) and IL-13 (Figure17B ).Figure17A shows the percent inhibition of TSLP signaling via the TSLP receptor complex expressed on STAT5-HEK-293 reporter cells when treated with one of the heterodimeric bispecific antibodies.Figure17B shows the percent inhibition of IL-13 signaling via the IL-13 receptor complex expressed on HEK-293-SEAP reporter cells when treated with one of the heterodimeric bispecific antibodies.IC90 values for each antibody against the antigen were calculated based on the dose-dependent binding inhibition curves. Data were analyzed and presented using GraphPad. Ab, antibody; Conc., concentration.

圖18A-18C顯示了僅以TSLP (圖18A)、僅以IL-13 (圖18B)或TSLP及IL-13 (圖18C)處理的PBMC中CCL17的產量，係因圖14所示之PX128-JT7及PX128-JT11異二聚體雙特異性抗體的劑量增加所致，如藉由ELISA試驗的測量。圖18A顯示了以單獨的0.5 ng/ml TSLP處理的CCL17產量。參考抗IL-13 Ab. #2、參考TSLP×IL-13 Ab. #3、參考TSLP×IL-13×HSA Ab. #4及參考抗TSLP Ab. #5作為對照。僅以TSLP (圖18A)、僅以IL-13 (圖18B)或TSLP及IL-13 (圖18C)處理細胞，並以不處理抗體的細胞作為陰性對照。Figures18A-18C show the production of CCL17 in PBMCs treated with TSLP alone (Figure18A ), IL-13 alone (Figure18B ), or TSLP and IL-13 (Figure18C ), as measured by ELISA, in response to increasing doses of the PX128-JT7 and PX128-JT11 heterodimeric bispecific antibodies shown inFigure14.Figure18A shows CCL17 production following treatment with 0.5 ng/ml TSLP alone. Reference anti-IL-13 Ab #2, reference TSLP × IL-13 Ab #3, reference TSLP × IL-13 × HSA Ab #4, and reference anti-TSLP Ab #5 served as controls. Cells were treated with TSLP alone (FIG.18A ), IL-13 alone (FIG.18B ), or TSLP and IL-13 (FIG.18C ). Cells without antibody treatment served as negative controls.

圖19顯示了(圖14所示之)示例性PX128-JT11異二聚體雙特異性抗體的體內穩定性半衰期，其係藉由靜脈內推注或皮下注射至食蟹獼猴後隨時間變化的體內抗體濃度來呈現。參考TSLP×IL-13×HSA Ab. #4作為對照。針對每種抗體的每個濃度測試兩隻動物(n=2/組)。Figure19 shows the in vivo stability half-life of the exemplary PX128-JT11 heterodimeric bispecific antibody (shown inFigure14 ), as measured by time-dependent antibody concentrations following intravenous bolus or subcutaneous injection in cynomolgus macaques. TSLP × IL-13 × HSA Ab #4 was used as a reference control. Two animals (n = 2/group) were tested at each concentration of each antibody.

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TW202525846A_113131850_SEQL.xmlTW202525846A_113131850_SEQL.xml

Claims (57)

Translated fromChinese
一種多特異性構築體，其包含： (1) 特異性地結合至介白素-13 (IL-13)的第一抗體部分，以及 (2) 特異性地結合至第二抗原的第二抗體部分。A multispecific construct comprising:(1) a first antibody portion that specifically binds to interleukin-13 (IL-13), and(2) a second antibody portion that specifically binds to a second antigen.如請求項1之多特異性構築體，其中該第一抗體部分包含重鏈可變區(VH1)及輕鏈可變區(VL1)，其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多3個胺基酸變異的變體，(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多3個胺基酸變異的變體，以及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多3個胺基酸變異的變體，且該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1或其含有至多3個胺基酸變異的變體，(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多3個胺基酸變異的變體，以及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多3個胺基酸變異的變體。The multispecific construct of claim 1, wherein the first antibody portion comprises a heavy chain variable region (VH1) and a light chain variable region (VL1), wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66 or a variant thereof containing up to 3 amino acid variations, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67 or a variant thereof containing up to 3 amino acid variations, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68 or a variant thereof containing up to 3 amino acid variations, and the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70 or a variant thereof containing up to 3 amino acid variations, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67 or a variant thereof containing up to 3 amino acid variations, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68 or a variant thereof containing up to 3 amino acid variations. (iii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71 or a variant thereof containing up to 3 amino acid variations, and (iv) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72 or a variant thereof containing up to 3 amino acid variations.如請求項2之多特異性構築體，其中該VH1包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1，(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2，以及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3，且該VL1包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1，(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2，以及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3。The multispecific construct of claim 2, wherein the VH1 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, and the VL1 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70, (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72.如請求項2或3之多特異性構築體，其中該VH1含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80%序列同一性的變體，且該VL1含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80%序列同一性的變體。The multispecific construct of claim 2 or 3, wherein the VH1 comprises the amino acid sequence of SEQ ID NO: 65 or a variant thereof having at least about 80% sequence identity with SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69 or a variant thereof having at least about 80% sequence identity with SEQ ID NO: 69.如請求項4之多特異性構築體，其中該VH1含有SEQ ID NO:65之胺基酸序列，且該VL1含有SEQ ID NO:69之胺基酸序列。The multispecific construct of claim 4, wherein the VH1 comprises the amino acid sequence of SEQ ID NO: 65, and the VL1 comprises the amino acid sequence of SEQ ID NO: 69.如請求項1至5中任一項之多特異性構築體，其中該第一抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、sdAb及scFv。The multispecific construct of any one of claims 1 to 5, wherein the first antibody portion is selected from the group consisting of a full-length antibody, Fab, Fab', F(ab')2 , sdAb, and scFv.如請求項6之多特異性構築體，其中該第一抗體部分為scFv (抗IL-13 scFv)。The multispecific construct of claim 6, wherein the first antibody portion is scFv (anti-IL-13 scFv).如請求項6之多特異性構築體，其中該第一抗體部分為Fab (抗IL-13 Fab)。The multispecific construct of claim 6, wherein the first antibody portion is Fab (anti-IL-13 Fab).如請求項6之多特異性構築體，其中該第一抗體部分為全長抗體(抗IL-13全長抗體)。The multispecific construct of claim 6, wherein the first antibody portion is a full-length antibody (anti-IL-13 full-length antibody).如請求項1至9中任一項之多特異性構築體，其中該第二抗原為胸腺基質淋巴球生成素(thymic stromal lymphopoietin，TSLP)。The multispecific construct of any one of claims 1 to 9, wherein the second antigen is thymic stromal lymphopoietin (TSLP).如請求項10之多特異性構築體，其中該第二抗體部分包含重鏈可變區(VH2)及輕鏈可變區(VL2)，其中： (a)該VH2包含(i)含有SEQ ID NO:2之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:3之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3； (b)該VH2包含(i)含有SEQ ID NO:12之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:13之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:4之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:6之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:7之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:8之胺基酸序列的CDR-L3； (c)該VH2包含(i)含有SEQ ID NO:20之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:21之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:22之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:24之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:25之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:26之胺基酸序列的CDR-L3； (d)該VH2包含(i)含有SEQ ID NO:28之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:29之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:30之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:32之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:33之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:34之胺基酸序列的CDR-L3； (e)該VH2包含(i)含有SEQ ID NO:36之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:37之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:38之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:40之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:41之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:42之胺基酸序列的CDR-L3；或 (f)該VH2包含(i)含有SEQ ID NO:44之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:45之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:46之胺基酸序列的CDR-H3；且該VL2包含(i)含有SEQ ID NO:48之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:49之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:50之胺基酸序列的CDR-L3。The multispecific construct of claim 10, wherein the second antibody portion comprises a heavy chain variable region (VH2) and a light chain variable region (VL2), wherein:(a) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:2; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:3; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:4; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:6; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:7; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:8;(b) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:12; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:3; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:4. NO:13; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:4; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:6; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:7; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:8;(c) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:20; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:21; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:22; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:24; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:25; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: NO:26;(d) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:28; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:29; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:30; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:32; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:33; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:34;(e) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:36; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:37; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:38. (i) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:38; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:40; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:41; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:42; or(f) the VH2 comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO:44; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO:45; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO:46; and the VL2 comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO:48; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO:49; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO:50.如請求項11之多特異性構築體，其中： (a)該VH2含有SEQ ID NO:1之胺基酸序列，或與SEQ ID NO:1具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:5之胺基酸序列，或與SEQ ID NO:5具有至少約80%序列同一性的胺基酸序列； (b)該VH2含有SEQ ID NO:9之胺基酸序列，或與SEQ ID NO:9具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:10之胺基酸序列，或與SEQ ID NO:10具有至少約80%序列同一性的胺基酸序列； (c)該VH2含有SEQ ID NO:11之胺基酸序列，或與SEQ ID NO:11具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:15之胺基酸序列，或與SEQ ID NO:15具有至少約80%序列同一性的胺基酸序列； (d)該VH2含有SEQ ID NO:19之胺基酸序列，或與SEQ ID NO:19具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:23之胺基酸序列，或與SEQ ID NO:23具有至少約80%序列同一性的胺基酸序列； (e)該VH2含有SEQ ID NO:27之胺基酸序列，或與SEQ ID NO:27具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:31之胺基酸序列，或與SEQ ID NO:31具有至少約80%序列同一性的胺基酸序列； (f)該VH2含有SEQ ID NO:35之胺基酸序列，或與SEQ ID NO:35具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:39之胺基酸序列，或與SEQ ID NO:39具有至少約80%序列同一性的胺基酸序列； (g)該VH2含有SEQ ID NO:43之胺基酸序列，或與SEQ ID NO:43具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:47之胺基酸序列，或與SEQ ID NO:47具有至少約80%序列同一性的胺基酸序列； (h)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (i)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (j)該VH2含有SEQ ID NO:56之胺基酸序列，或與SEQ ID NO:56具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (k)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列； (l)該VH2含有SEQ ID NO:56之胺基酸序列，或與SEQ ID NO:56具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列； (m)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列； (n)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:51之胺基酸序列，或與SEQ ID NO:51具有至少約80%序列同一性的胺基酸序列； (o)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (p)該VH2含有SEQ ID NO:58之胺基酸序列，或與SEQ ID NO:58具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (q)該VH2含有SEQ ID NO:60之胺基酸序列，或與SEQ ID NO:60具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (r)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (s)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (t)該VH2含有SEQ ID NO:57之胺基酸序列，或與SEQ ID NO:57具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (u)該VH2含有SEQ ID NO:58之胺基酸序列，或與SEQ ID NO:58具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (v)該VH2含有SEQ ID NO:62之胺基酸序列，或與SEQ ID NO:62具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (w)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:53之胺基酸序列，或與SEQ ID NO:53具有至少約80%序列同一性的胺基酸序列； (x)該VH2含有SEQ ID NO:55之胺基酸序列，或與SEQ ID NO:55具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (y)該VH2含有SEQ ID NO:61之胺基酸序列，或與SEQ ID NO:61具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (z)該VH2含有SEQ ID NO:62之胺基酸序列，或與SEQ ID NO:62具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (aa)該VH2含有SEQ ID NO:64之胺基酸序列，或與SEQ ID NO:64具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:54之胺基酸序列，或與SEQ ID NO:54具有至少約80%序列同一性的胺基酸序列； (bb)該VH2含有SEQ ID NO:63之胺基酸序列，或與SEQ ID NO:63具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:52之胺基酸序列，或與SEQ ID NO:52具有至少約80%序列同一性的胺基酸序列； (cc)該VH2含有SEQ ID NO:188之胺基酸序列，或與SEQ ID NO:188具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:189之胺基酸序列，或與SEQ ID NO:189具有至少約80%序列同一性的胺基酸序列； (dd)該VH2含有SEQ ID NO:188之胺基酸序列，或與SEQ ID NO:188具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:190之胺基酸序列，或與SEQ ID NO:190具有至少約80%序列同一性的胺基酸序列；或 (ee)該VH2含有SEQ ID NO:228之胺基酸序列，或與SEQ ID NO:228具有至少約80%序列同一性的胺基酸序列，且該VL2含有SEQ ID NO:190之胺基酸序列，或與SEQ ID NO:190具有至少約80%序列同一性的胺基酸序列。The multispecific construct of claim 11, wherein:(a) the VH2 comprises the amino acid sequence of SEQ ID NO:1, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:1, and the VL2 comprises the amino acid sequence of SEQ ID NO:5, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:5;(b) the VH2 comprises the amino acid sequence of SEQ ID NO:9, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:9, and the VL2 comprises the amino acid sequence of SEQ ID NO:10, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:10;(c) the VH2 comprises the amino acid sequence of SEQ ID NO:11, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:11, and the VL2 comprises the amino acid sequence of SEQ ID NO:15, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:15. NO:15;(d) the VH2 comprises the amino acid sequence of SEQ ID NO:19, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:19, and the VL2 comprises the amino acid sequence of SEQ ID NO:23, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:23;(e) the VH2 comprises the amino acid sequence of SEQ ID NO:27, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:27, and the VL2 comprises the amino acid sequence of SEQ ID NO:31, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:31;(f) the VH2 comprises the amino acid sequence of SEQ ID NO:35, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:35, and the VL2 comprises the amino acid sequence of SEQ ID NO: NO:39, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:39;(g) the VH2 comprises the amino acid sequence of SEQ ID NO:43, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:43, and the VL2 comprises the amino acid sequence of SEQ ID NO:47, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:47;(h) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:53;(i) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: NO:63 has an amino acid sequence with at least about 80% sequence identity, and the VL2 contains the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:54;(j) the VH2 contains the amino acid sequence of SEQ ID NO:56, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:56, and the VL2 contains the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:52;(k) the VH2 contains the amino acid sequence of SEQ ID NO:55, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:55, and the VL2 contains the amino acid sequence of SEQ ID NO:51, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:51;(l) the VH2 contains the amino acid sequence of SEQ ID NO: (i) wherein the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:51, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:51; (ii) wherein the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:51, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:51; (o) the VH2 comprises the amino acid sequence of SEQ ID NO:55, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:55, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:52;(p) the VH2 comprises the amino acid sequence of SEQ ID NO:58, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:58, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:52;(q) the VH2 comprises the amino acid sequence of SEQ ID NO:60, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:60, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: NO:52; (r) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:52; (s) the VH2 comprises the amino acid sequence of SEQ ID NO:55, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:55, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:53; (t) the VH2 comprises the amino acid sequence of SEQ ID NO:57, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:57, and the VL2 comprises the amino acid sequence of SEQ ID NO:53; NO:53, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:53;(u) the VH2 comprises the amino acid sequence of SEQ ID NO:58, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:58, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:53;(v) the VH2 comprises the amino acid sequence of SEQ ID NO:62, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:62, and the VL2 comprises the amino acid sequence of SEQ ID NO:53, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:53;(w) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO:65. NO:64 has an amino acid sequence with at least about 80% sequence identity, and the VL2 contains the amino acid sequence of SEQ ID NO:53, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:53;(x) the VH2 contains the amino acid sequence of SEQ ID NO:55, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:55, and the VL2 contains the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:54;(y) the VH2 contains the amino acid sequence of SEQ ID NO:61, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:61, and the VL2 contains the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:54;(z) the VH2 contains the amino acid sequence of SEQ ID NO: (aa) the VH2 comprises the amino acid sequence of SEQ ID NO:64, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:64, and the VL2 comprises the amino acid sequence of SEQ ID NO:54, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:54; (bb) the VH2 comprises the amino acid sequence of SEQ ID NO:63, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:63, and the VL2 comprises the amino acid sequence of SEQ ID NO:52, or an amino acid sequence with at least about 80% sequence identity to SEQ ID NO:52; (cc) the VH2 comprises the amino acid sequence of SEQ ID NO: 188, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 188, and the VL2 comprises the amino acid sequence of SEQ ID NO: 189, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 189;(dd) the VH2 comprises the amino acid sequence of SEQ ID NO: 188, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 188, and the VL2 comprises the amino acid sequence of SEQ ID NO: 190, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 190; or(ee) the VH2 comprises the amino acid sequence of SEQ ID NO: 228, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: 228, and the VL2 comprises the amino acid sequence of SEQ ID NO: 190, or an amino acid sequence having at least about 80% sequence identity to SEQ ID NO: NO:190 has an amino acid sequence with at least about 80% sequence identity.如請求項1至12中任一項之多特異性構築體，其中該第二抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、sdAb及scFv。The multispecific construct of any one of claims 1 to 12, wherein the second antibody portion is selected from the group consisting of a full-length antibody, Fab, Fab', F(ab')2 , sdAb, and scFv.如請求項13之多特異性構築體，其中該第二抗體部分為全長抗體。The multispecific construct of claim 13, wherein the second antibody portion is a full-length antibody.如請求項13之多特異性構築體，其中該第二抗體部分為scFv。The multispecific construct of claim 13, wherein the second antibody portion is a scFv.如請求項13之多特異性構築體，其中該第二抗體部分為Fab。The multispecific construct of claim 13, wherein the second antibody portion is Fab.如請求項16之多特異性構築體，其中該第二抗體部分為抗TSLP Fab，且其中該抗TSLP Fab包含含有SEQ ID NO:109之胺基酸序列的第一多肽及含有SEQ ID NO:103之胺基酸序列的第二多肽。The multispecific construct of claim 16, wherein the second antibody portion is an anti-TSLP Fab, and wherein the anti-TSLP Fab comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 109 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 103.如請求項1至5、8及10至17中任一項之多特異性構築體，其中該第一抗體部分包含重鏈(H1)及輕鏈(L1)，其中該H1包含VH1及H1-CH1，其中該L1包含VL1及L1-CL，且其中： (i)該H1包含位於位置170、183及185處的取代(EU編號)，且該L1包含位於位置135處的取代(EU編號)；及/或 (ii)該H1包含位於位置126及220處的取代(EU編號)，且該L1包含位於位置124及214處的取代(EU編號)。The multispecific construct of any one of claims 1 to 5, 8, and 10 to 17, wherein the first antibody portion comprises a heavy chain (H1) and a light chain (L1), wherein the H1 comprises VH1 and H1-CH1, wherein the L1 comprises VL1 and L1-CL, and wherein:(i) the H1 comprises substitutions at positions 170, 183, and 185 (EU numbering), and the L1 comprises a substitution at position 135 (EU numbering); and/or(ii) the H1 comprises substitutions at positions 126 and 220 (EU numbering), and the L1 comprises substitutions at positions 124 and 214 (EU numbering).如請求項18之多特異性構築體，其中該H1包含位於位置F170、S183及V185處的取代(EU編號)，且該L1包含位於位置L135處的取代(EU編號)。The multispecific construct of claim 18, wherein the H1 comprises substitutions at positions F170, S183, and V185 (EU numbering), and the L1 comprises a substitution at position L135 (EU numbering).如請求項19之多特異性構築體， (i)其中該F170取代為F170I或F170V、該S183取代為S183L或S183I且該V185取代為V185L；及/或 (ii)其中該L135取代為L135F。The multispecific construct of claim 19,(i) wherein F170 is substituted with F170I or F170V, S183 is substituted with S183L or S183I, and V185 is substituted with V185L; and/or(ii) wherein L135 is substituted with L135F.如請求項20之多特異性構築體，其中： (a)該H1包含F170I、S183L及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)；或 (b)該H1包含F170V、S183I及V185L取代(EU編號)，且該L1包含L135F取代(EU編號)。The multispecific construct of claim 20, wherein:(a) the H1 comprises F170I, S183L, and V185L substitutions (EU numbering), and the L1 comprises L135F substitution (EU numbering); or(b) the H1 comprises F170V, S183I, and V185L substitutions (EU numbering), and the L1 comprises L135F substitution (EU numbering).如請求項18至21中任一項之多特異性構築體，其中該L1係衍生自λ輕鏈。The multispecific construct of any one of claims 18 to 21, wherein L1 is derived from a lambda light chain.如請求項22之多特異性構築體，其中該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置E124及C214處的取代(EU編號)。The multispecific construct of claim 22, wherein the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions E124 and C214 (EU numbering).如請求項23之多特異性構築體，其中該H1包含F126C及C220S取代(EU編號)，且該L1包含E124C及C214S取代(EU編號)。The multispecific construct of claim 23, wherein the H1 comprises F126C and C220S substitutions (EU numbering), and the L1 comprises E124C and C214S substitutions (EU numbering).如請求項18至21中任一項之多特異性構築體，其中該L1係衍生自κ輕鏈。The multispecific construct of any one of claims 18 to 21, wherein L1 is derived from a kappa light chain.如請求項25之多特異性構築體，其中該H1包含位於位置F126及C220處的取代(EU編號)，且該L1包含位於位置Q124及C214處的取代(EU編號)。The multispecific construct of claim 25, wherein the H1 comprises substitutions at positions F126 and C220 (EU numbering), and the L1 comprises substitutions at positions Q124 and C214 (EU numbering).如請求項26之多特異性構築體，其中該H1包含F126C及C220S取代(EU編號)，且該L1包含Q124C及C214S取代(EU編號)。The multispecific construct of claim 26, wherein the H1 comprises F126C and C220S substitutions (EU numbering), and the L1 comprises Q124C and C214S substitutions (EU numbering).如請求項18至27中任一項之多特異性構築體，其中 (i)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代； (ii)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代； (iii)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含Q124C、L135F及C214S取代；或 (iv)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含Q124C、L135F及C214S取代；且 其中該位置係根據EU編號。The multispecific construct of any one of claims 18 to 27, wherein:(i) H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and L1 comprises E124C, L135F, and C214S substitutions;(ii) H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and L1 comprises E124C, L135F, and C214S substitutions;(iii) H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and L1 comprises Q124C, L135F, and C214S substitutions; or (iv) the H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and the L1 comprises Q124C, L135F, and C214S substitutions; andwherein the positions are according to EU numbering.如請求項1至8、10至13及15至28中任一項之多特異性構築體，其中該多特異性構築體更包含Fc域，其中該Fc域包含第一次單元及第二次單元。The multispecific construct of any one of claims 1 to 8, 10 to 13, and 15 to 28, wherein the multispecific construct further comprises an Fc domain, wherein the Fc domain comprises a first subunit and a second subunit.如請求項29之多特異性構築體，其中該Fc域係衍生自IgG1。The multispecific construct of claim 29, wherein the Fc domain is derived from IgG1.如請求項29或30之多特異性構築體，其中： (i)該Fc域之每一次單元包含L234A及L235A取代(EU編號)； (ii)該Fc域之每一次單元包含M428L及N434S取代(EU編號)； (iii)該Fc域之每一次單元包含M252Y、S254T及T256E取代(EU編號)；及/或 (iv) (a)該Fc域之第一次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第二次單元包含T366W取代(EU編號)；或 (b)該Fc域之第二次單元包含T366S、L368A及Y407V取代(EU編號)，且該Fc域之第一次單元包含T366W取代(EU編號)。The multispecific construct of claim 29 or 30, wherein:(i) each subunit of the Fc domain comprises L234A and L235A substitutions (EU numbering);(ii) each subunit of the Fc domain comprises M428L and N434S substitutions (EU numbering);(iii) each subunit of the Fc domain comprises M252Y, S254T, and T256E substitutions (EU numbering); and/or(iv) (a) the first subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the second subunit of the Fc domain comprises T366W substitution (EU numbering); or(b) the second subunit of the Fc domain comprises T366S, L368A, and Y407V substitutions (EU numbering), and the first subunit of the Fc domain comprises T366W substitution (EU numbering).如請求項31之多特異性構築體，其中： (i)該Fc域之第一次單元含有SEQ ID NO:80之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:81之胺基酸序列； (ii)該Fc域之第一次單元含有SEQ ID NO:81之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:80之胺基酸序列； (iii)該Fc域之第一次單元含有SEQ ID NO:95之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:96之胺基酸序列；或 (iv)該Fc域之第一次單元含有SEQ ID NO:96之胺基酸序列，且該Fc域之第二次單元含有SEQ ID NO:95之胺基酸序列。The multispecific construct of claim 31, wherein:(i) the first subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:80, and the second subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:81;(ii) the first subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:81, and the second subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:80;(iii) the first subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:95, and the second subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:96; or(iv) the first subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:96, and the second subunit of the Fc domain comprises the amino acid sequence of SEQ ID NO:95.如請求項1至5、10至12及18至32中任一項之多特異性構築體，其中該多特異性構築體為異二聚體雙特異性抗體，其包含： i)含有VL1及L1-CL的第一輕鏈(L1)； ii)含有VH1、H1-CH1及Fc域之第一次單元的第一重鏈(H1)； iii)含有VH2、H2-CH1及該Fc域之第二次單元的第二重鏈(H2)；及 iv)含有VL2及L2-CL的第二輕鏈(L2)；且 其中H1及L1形成抗IL-13抗體部分，且H2及L2形成該第二抗體部分。The multispecific construct of any one of claims 1 to 5, 10 to 12, and 18 to 32, wherein the multispecific construct is a heterodimeric bispecific antibody comprising:i) a first light chain (L1) comprising VL1 and L1-CL;ii) a first heavy chain (H1) comprising a first subunit comprising VH1, H1-CH1, and an Fc domain;iii) a second heavy chain (H2) comprising a second subunit comprising VH2, H2-CH1, and the Fc domain; andiv) a second light chain (L2) comprising VL2 and L2-CL; andwherein H1 and L1 form the anti-IL-13 antibody portion, and H2 and L2 form the second antibody portion.如請求項33之多特異性構築體，其中： (i)該Fc域係衍生自人類IgG1；及/或 (ii)其中該L1-CL係衍生自人類λ輕鏈。The multispecific construct of claim 33, wherein:(i) the Fc domain is derived from human IgG1; and/or(ii) the L1-CL is derived from a human lambda light chain.如請求項33或34之多特異性構築體，其中： (a)該H1包含F170I、S183L及V185L取代，且該L1包含L135F取代； (b)該H1包含F170V、S183I及V185L取代，且該L1包含L135F取代； (c)該H1包含F126C、F170I、S183L、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代； (d)該H1包含F126C、F170V、S183I、V185L及C220S取代，且該L1包含E124C、L135F及C214S取代； (e)該H1包含F126C及C220S取代，且該L1包含E124C及C214S取代； (f)該H1包含F170I、S183L、V185L、T366S、L368A及Y407V取代，該L1包含L135F取代，且該H2包含T366W取代； (g)該H1包含F170I、S183L、V185L及T366W取代，該L1包含L135F取代，且該H2包含T366S、L368A及Y407V取代； (h)該H1包含F170V、S183I、V185L、T366S、L368A及Y407V取代，該L1包含L135F取代，且該H2包含T366W取代； (i)該H1包含F170V、S183I、V185L及T366W取代，該L1包含L135F取代，且該H2包含T366S、L368A及Y407V取代； (j)該H1包含F126C、F170I、S183L、V185L、C220S、T366S、L368A及Y407V取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366W取代； (k)該H1包含F126C、F170I、S183L、V185L、C220S及T366W取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366S、L368A及Y407V取代； (l)該H1包含F126C、F170V、S183I、V185L、C220S、T366S、L368A及Y407V取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366W取代； (m)該H1包含F126C、F170V、S183I、V185L、C220S及T366W取代，該L1包含E124C、L135F及C214S取代，且該H2包含T366S、L368A及Y407V取代； (n)該H1包含F126C、C220S、T366S、L368A及Y407V取代，該L1包含E124C及C214S取代，且該H2包含T366W取代； (o)該H1包含F126C、C220S及T366W取代，該L1包含E124C及C214S取代，且該H2包含T366S、L368A及Y407V取代； (p)該H1包含F170I、S183L、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代； (q)該H1包含F170I、S183L、V185L、L234A、L235A、T366W、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (r)該H1包含F170V、S183I、V185L、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代； (s)該H1包含F170V、S183I、V185L、L234A、L235A、T366W、M428L及N434S取代，該L1包含L135F取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (t)該H1包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代； (u)該H1包含F126C、F170I、S183L、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (v)該H1包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代； (w)該H1包含F126C、F170V、S183I、V185L、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C、L135F及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代； (x)該H1包含F126C、C220S、L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代，該L1包含E124C及C214S取代，且該H2包含L234A、L235A、T366W、M428L及N434S取代；或 (y)該H1包含F126C、C220S、L234A、L235A、T366W、M428L及N434S取代，該L1包含E124C及C214S取代，且該H2包含L234A、L235A、T366S、L368A、Y407V、M428L及N434S取代；且 其中該胺基酸位置係根據EU編號。The multispecific construct of claim 33 or 34, wherein:(a) H1 comprises F170I, S183L, and V185L substitutions, and L1 comprises L135F substitutions;(b) H1 comprises F170V, S183I, and V185L substitutions, and L1 comprises L135F substitutions;(c) H1 comprises F126C, F170I, S183L, V185L, and C220S substitutions, and L1 comprises E124C, L135F, and C214S substitutions;(d) H1 comprises F126C, F170V, S183I, V185L, and C220S substitutions, and L1 comprises E124C, L135F, and C214S substitutions; (e) H1 comprises F126C and C220S substitutions, and L1 comprises E124C and C214S substitutions;(f) H1 comprises F170I, S183L, V185L, T366S, L368A, and Y407V substitutions, L1 comprises L135F substitution, and H2 comprises T366W substitution;(g) H1 comprises F170I, S183L, V185L, and T366W substitutions, L1 comprises L135F substitution, and H2 comprises T366S, L368A, and Y407V substitutions;(h) H1 comprises F170V, S183I, V185L, T366S, L368A, and Y407V substitutions, L1 comprises L135F substitution, and H2 comprises T366W substitution; (i) H1 comprises F170V, S183I, V185L, and T366W substitutions, L1 comprises L135F substitutions, and H2 comprises T366S, L368A, and Y407V substitutions;(j) H1 comprises F126C, F170I, S183L, V185L, C220S, T366S, L368A, and Y407V substitutions, L1 comprises E124C, L135F, and C214S substitutions, and H2 comprises T366W substitution; (k) the H1 comprises F126C, F170I, S183L, V185L, C220S, and T366W substitutions, the L1 comprises E124C, L135F, and C214S substitutions, and the H2 comprises T366S, L368A, and Y407V substitutions;(l) the H1 comprises F126C, F170V, S183I, V185L, C220S, T366S, L368A, and Y407V substitutions, the L1 comprises E124C, L135F, and C214S substitutions, and the H2 comprises T366W substitutions; (m) the H1 comprises F126C, F170V, S183I, V185L, C220S, and T366W substitutions, the L1 comprises E124C, L135F, and C214S substitutions, and the H2 comprises T366S, L368A, and Y407V substitutions;(n) the H1 comprises F126C, C220S, T366S, L368A, and Y407V substitutions, the L1 comprises E124C and C214S substitutions, and the H2 comprises T366W substitutions;(o) the H1 comprises F126C, C220S, and T366W substitutions, the L1 comprises E124C and C214S substitutions, and the H2 comprises T366S, L368A, and Y407V substitutions; (p) H1 comprises F170I, S183L, V185L, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, L1 comprises L135F substitutions, and H2 comprises L234A, L235A, T366W, M428L, and N434S substitutions;(q) H1 comprises F170I, S183L, V185L, L234A, L235A, T366W, M428L, and N434S substitutions, L1 comprises L135F substitutions, and H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions; (r) the H1 comprises F170V, S183I, V185L, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, the L1 comprises L135F substitutions, and the H2 comprises L234A, L235A, T366W, M428L, and N434S substitutions;(s) the H1 comprises F170V, S183I, V185L, L234A, L235A, T366W, M428L, and N434S substitutions, the L1 comprises L135F substitutions, and the H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions; (t) the H1 comprises F126C, F170I, S183L, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises L234A, L235A, T366W, M428L and N434S substitutions; (u) the H1 comprises F126C, F170I, S183L, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (v) the H1 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises L234A, L235A, T366W, M428L and N434S substitutions; (w) the H1 comprises F126C, F170V, S183I, V185L, C220S, L234A, L235A, T366W, M428L and N434S substitutions, the L1 comprises E124C, L135F and C214S substitutions, and the H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L and N434S substitutions; (x) H1 comprises F126C, C220S, L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions, L1 comprises E124C and C214S substitutions, and H2 comprises L234A, L235A, T366W, M428L, and N434S substitutions; or(y) H1 comprises F126C, C220S, L234A, L235A, T366W, M428L, and N434S substitutions, L1 comprises E124C and C214S substitutions, and H2 comprises L234A, L235A, T366S, L368A, Y407V, M428L, and N434S substitutions; and The amino acid positions are based on EU numbering.如請求項35之多特異性構築體，其中： (a)該H1含有SEQ ID NO:87之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列； (b)該H1含有SEQ ID NO:88之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列； (c)該H1含有SEQ ID NO:89之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列； (d)該H1含有SEQ ID NO:90之胺基酸序列，該L1含有SEQ ID NO:173或202之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列； (e)該H1含有SEQ ID NO:91之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列； (f)該H1含有SEQ ID NO:92之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列； (g)該H1含有SEQ ID NO:93之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列； (h)該H1含有SEQ ID NO:94之胺基酸序列，該L1含有SEQ ID NO:174或203之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列； (i)該H1含有SEQ ID NO:198之胺基酸序列，該L1含有SEQ ID NO:204或205之胺基酸序列，且該H2含有SEQ ID NO:85之胺基酸序列；或 (j)該H1含有SEQ ID NO:199之胺基酸序列，該L1含有SEQ ID NO:204或205之胺基酸序列，且該H2含有SEQ ID NO:86之胺基酸序列。The multispecific construct of claim 35, wherein:(a) the H1 contains the amino acid sequence of SEQ ID NO:87, the L1 contains the amino acid sequence of SEQ ID NO:173 or 202, and the H2 contains the amino acid sequence of SEQ ID NO:85;(b) the H1 contains the amino acid sequence of SEQ ID NO:88, the L1 contains the amino acid sequence of SEQ ID NO:173 or 202, and the H2 contains the amino acid sequence of SEQ ID NO:86;(c) the H1 contains the amino acid sequence of SEQ ID NO:89, the L1 contains the amino acid sequence of SEQ ID NO:173 or 202, and the H2 contains the amino acid sequence of SEQ ID NO:85;(d) the H1 contains the amino acid sequence of SEQ ID NO:90, the L1 contains the amino acid sequence of SEQ ID NO:173 or 202, and the H2 contains the amino acid sequence of SEQ ID NO: (e) the H1 contains the amino acid sequence of SEQ ID NO:91, the L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H2 contains the amino acid sequence of SEQ ID NO:85; (f) the H1 contains the amino acid sequence of SEQ ID NO:92, the L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H2 contains the amino acid sequence of SEQ ID NO:86; (g) the H1 contains the amino acid sequence of SEQ ID NO:93, the L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H2 contains the amino acid sequence of SEQ ID NO:85; (h) the H1 contains the amino acid sequence of SEQ ID NO:94, the L1 contains the amino acid sequence of SEQ ID NO:174 or 203, and the H2 contains the amino acid sequence of SEQ ID NO:86; (i) the H1 contains the amino acid sequence of SEQ ID NO: 198, the L1 contains the amino acid sequence of SEQ ID NO: 204 or 205, and the H2 contains the amino acid sequence of SEQ ID NO: 85; or(j) the H1 contains the amino acid sequence of SEQ ID NO: 199, the L1 contains the amino acid sequence of SEQ ID NO: 204 or 205, and the H2 contains the amino acid sequence of SEQ ID NO: 86.如請求項33至36中任一項之多特異性構築體，其中該L2-CL係衍生自人類κ輕鏈。The multispecific construct of any one of claims 33 to 36, wherein the L2-CL is derived from a human kappa light chain.如請求項37之多特異性構築體，其中該第二抗原為TSLP，且其中： (a)該H1含有SEQ ID NO:139之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (b)該H1含有SEQ ID NO:138之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (c)該H1含有SEQ ID NO:141之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (d)該H1含有SEQ ID NO:140之胺基酸序列，該L1含有SEQ ID NO:122或207之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (e)該H1含有SEQ ID NO:126之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (f)該H1含有SEQ ID NO:127之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (g)該H1含有SEQ ID NO:128之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (h)該H1含有SEQ ID NO:129之胺基酸序列，該L1含有SEQ ID NO:120或208之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列； (i)該H1含有SEQ ID NO:200之胺基酸序列，該L1含有SEQ ID NO:121或206之胺基酸序列，該H2含有SEQ ID NO:136之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列；或 (j)該H1含有SEQ ID NO:201之胺基酸序列，該L1含有SEQ ID NO:121或206之胺基酸序列，該H2含有SEQ ID NO:137之胺基酸序列，且該L2含有SEQ ID NO:103之胺基酸序列。The multispecific construct of claim 37, wherein the second antigen is TSLP, and wherein:(a) the H1 contains the amino acid sequence of SEQ ID NO:139, the L1 contains the amino acid sequence of SEQ ID NO:122 or 207, the H2 contains the amino acid sequence of SEQ ID NO:136, and the L2 contains the amino acid sequence of SEQ ID NO:103;(b) the H1 contains the amino acid sequence of SEQ ID NO:138, the L1 contains the amino acid sequence of SEQ ID NO:122 or 207, the H2 contains the amino acid sequence of SEQ ID NO:137, and the L2 contains the amino acid sequence of SEQ ID NO:103;(c) the H1 contains the amino acid sequence of SEQ ID NO:141, the L1 contains the amino acid sequence of SEQ ID NO:122 or 207, and the H2 contains the amino acid sequence of SEQ ID NO:141. (d) the H1 contains the amino acid sequence of SEQ ID NO: 140, the L1 contains the amino acid sequence of SEQ ID NO: 122 or 207, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (e) the H1 contains the amino acid sequence of SEQ ID NO: 126, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (f) the H1 contains the amino acid sequence of SEQ ID NO: 127, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, and the H2 contains the amino acid sequence of SEQ ID NO: 137. (g) the H1 contains the amino acid sequence of SEQ ID NO: 128, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 136, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (h) the H1 contains the amino acid sequence of SEQ ID NO: 129, the L1 contains the amino acid sequence of SEQ ID NO: 120 or 208, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103; (i) the H1 contains the amino acid sequence of SEQ ID NO: 200, the L1 contains the amino acid sequence of SEQ ID NO: 121 or 206, and the H2 contains the amino acid sequence of SEQ ID NO: 137. (j) the H1 contains the amino acid sequence of SEQ ID NO: 201, the L1 contains the amino acid sequence of SEQ ID NO: 121 or 206, the H2 contains the amino acid sequence of SEQ ID NO: 137, and the L2 contains the amino acid sequence of SEQ ID NO: 103.一種分離的抗體構築體(抗IL-13抗體構築體)，其包含特異性地結合介白素-13 (IL-13)(抗IL-13抗體部分)的抗體部分，其中該抗IL-13抗體部分包含重鏈可變區(VH)及輕鏈可變區(VL)，且其中：該VH包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1或其含有至多3個胺基酸變異的變體；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2或其含有至多3個胺基酸變異的變體；及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3或其含有至多3個胺基酸變異的變體；且該VL包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1或其含有至多3個胺基酸變異的變體；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2或其含有至多3個胺基酸變異的變體；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3或其含有至多3個胺基酸變異的變體。An isolated antibody construct (anti-IL-13 antibody construct) comprising an antibody portion that specifically binds interleukin-13 (IL-13) (anti-IL-13 antibody portion), wherein the anti-IL-13 antibody portion comprises a heavy chain variable region (VH) and a light chain variable region (VL), and wherein: the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66, or a variant thereof containing up to 3 amino acid variations; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to 3 amino acid variations; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68, or a variant thereof containing up to 3 amino acid variations; and the VL comprises (i) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67, or a variant thereof containing up to 3 amino acid variations; (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70, or a variant thereof containing up to 3 amino acid variants; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71, or a variant thereof containing up to 3 amino acid variants; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72, or a variant thereof containing up to 3 amino acid variants.如請求項39之分離的抗IL-13抗體構築體，其中該VH包含(i)含有SEQ ID NO:66之胺基酸序列的CDR-H1；(ii)含有SEQ ID NO:67之胺基酸序列的CDR-H2；及(iii)含有SEQ ID NO:68之胺基酸序列的CDR-H3；且該VL包含(i)含有SEQ ID NO:70之胺基酸序列的CDR-L1；(ii)含有SEQ ID NO:71之胺基酸序列的CDR-L2；及(iii)含有SEQ ID NO:72之胺基酸序列的CDR-L3。The isolated anti-IL-13 antibody construct of claim 39, wherein the VH comprises (i) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 66; (ii) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 67; and (iii) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 68; and the VL comprises (i) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 70; (ii) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 71; and (iii) a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 72.如請求項39或40之分離的抗IL-13抗體構築體，其中該VH含有SEQ ID NO:65之胺基酸序列或其與SEQ ID NO:65具有至少約80%序列同一性的變體，且該VL含有SEQ ID NO:69之胺基酸序列或其與SEQ ID NO:69具有至少約80%序列同一性的變體。The isolated anti-IL-13 antibody construct of claim 39 or 40, wherein the VH comprises the amino acid sequence of SEQ ID NO: 65 or a variant thereof having at least about 80% sequence identity to SEQ ID NO: 65, and the VL comprises the amino acid sequence of SEQ ID NO: 69 or a variant thereof having at least about 80% sequence identity to SEQ ID NO: 69.如請求項41之分離的抗IL-13抗體構築體，其中該VH含有SEQ ID NO:65之胺基酸序列，且該VL含有SEQ ID NO:69之胺基酸序列。The isolated anti-IL-13 antibody construct of claim 41, wherein the VH comprises the amino acid sequence of SEQ ID NO: 65, and the VL comprises the amino acid sequence of SEQ ID NO: 69.如請求項39至42中任一項之分離的抗IL-13抗體，其中該抗IL-13抗體部分係選自由以下組成之群組：全長抗體、Fab、Fab’、F(ab’)2、雙抗體及scFv。The isolated anti-IL-13 antibody of any one of claims 39 to 42, wherein the anti-IL-13 antibody portion is selected from the group consisting of a full-length antibody, Fab, Fab', F(ab')2, a diabody, and a scFv.如請求項43之分離的抗IL-13抗體構築體，其中該抗IL-13抗體部分為scFv (抗IL-13 scFv)。The isolated anti-IL-13 antibody construct of claim 43, wherein the anti-IL-13 antibody portion is a scFv (anti-IL-13 scFv).如請求項44之分離的抗IL-13抗體構築體，其中該抗IL-13 scFv含有SEQ ID NO:108之胺基酸序列。The isolated anti-IL-13 antibody construct of claim 44, wherein the anti-IL-13 scFv comprises the amino acid sequence of SEQ ID NO: 108.如請求項43之分離的抗IL-13抗體構築體，其中該抗IL-13抗體部分為Fab (抗IL-13 Fab)。The isolated anti-IL-13 antibody construct of claim 43, wherein the anti-IL-13 antibody portion is Fab (anti-IL-13 Fab).如請求項46之分離的抗IL-13抗體構築體，其中該抗IL-13 Fab包含含有SEQ ID NO:124之胺基酸序列的第一多肽及含有SEQ ID NO:102或197之胺基酸序列的第二多肽。The isolated anti-IL-13 antibody construct of claim 46, wherein the anti-IL-13 Fab comprises a first polypeptide comprising the amino acid sequence of SEQ ID NO: 124 and a second polypeptide comprising the amino acid sequence of SEQ ID NO: 102 or 197.如請求項43之分離的抗IL-13抗體構築體，其中該抗IL-13抗體部分為全長抗體(抗IL-13全長抗體)。The isolated anti-IL-13 antibody construct of claim 43, wherein the anti-IL-13 antibody portion is a full-length antibody (anti-IL-13 full-length antibody).如請求項48之分離的抗IL-13抗體構築體，其中該抗IL-13全長抗體包含： i)兩條各自含有SEQ ID NO:125之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； ii)兩條各自含有SEQ ID NO:101之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈； iii)兩條各自含有SEQ ID NO:123之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈； iv)兩條各自含有SEQ ID NO:209之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； v)兩條各自含有SEQ ID NO:210之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:122或207之胺基酸序列的輕鏈； vi)兩條各自含有SEQ ID NO:211之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:121或206之胺基酸序列的輕鏈； vii)兩條各自含有SEQ ID NO:212之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； viii)兩條各自含有SEQ ID NO:213之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:120或208之胺基酸序列的輕鏈； ix)兩條各自含有SEQ ID NO:225之胺基酸序列的重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈；或 x)含有SEQ ID NO:130之胺基酸序列的第一重鏈、含有SEQ ID NO:131之胺基酸序列的第二重鏈及兩條各自含有SEQ ID NO:102或197之胺基酸序列的輕鏈。The isolated anti-IL-13 antibody construct of claim 48, wherein the full-length anti-IL-13 antibody comprises:i) two heavy chains each containing the amino acid sequence of SEQ ID NO: 125 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197;ii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 101 and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197;iii) two heavy chains each containing the amino acid sequence of SEQ ID NO: 123 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206;iv) two heavy chains each containing the amino acid sequence of SEQ ID NO: 209 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207; v) Two heavy chains each containing the amino acid sequence of SEQ ID NO: 210 and two light chains each containing the amino acid sequence of SEQ ID NO: 122 or 207;vi) Two heavy chains each containing the amino acid sequence of SEQ ID NO: 211 and two light chains each containing the amino acid sequence of SEQ ID NO: 121 or 206;vii) Two heavy chains each containing the amino acid sequence of SEQ ID NO: 212 and two light chains each containing the amino acid sequence of SEQ ID NO: 120 or 208;viii) Two heavy chains each containing the amino acid sequence of SEQ ID NO: 213 and two light chains each containing the amino acid sequence of SEQ ID NO: 120 or 208;ix) Two heavy chains each containing the amino acid sequence of SEQ ID NO: 225 and two light chains each containing the amino acid sequence of SEQ ID NO: A light chain containing the amino acid sequence of SEQ ID NO: 102 or 197; orx) a first heavy chain containing the amino acid sequence of SEQ ID NO: 130, a second heavy chain containing the amino acid sequence of SEQ ID NO: 131, and two light chains each containing the amino acid sequence of SEQ ID NO: 102 or 197.如請求項39至49中任一項之分離的抗IL-13抗體，其中該分離的抗IL-13抗體構築體係多特異性。The isolated anti-IL-13 antibody of any one of claims 39 to 49, wherein the isolated anti-IL-13 antibody construct is multispecific.如請求項50之分離的抗IL-13抗體構築體，其中該分離的抗IL-13抗體構築體更包含特異性地結合至第二抗原的第二抗體部分。The isolated anti-IL-13 antibody construct of claim 50, wherein the isolated anti-IL-13 antibody construct further comprises a second antibody portion that specifically binds to a second antigen.一種醫藥組成物，其包含(i)如請求項1至38中任一項之多特異性構築體，或如請求項39至51中任一項之分離的抗IL-13抗體；及(ii)醫藥上可接受之載體。A pharmaceutical composition comprising (i) the multispecific construct of any one of claims 1 to 38, or the isolated anti-IL-13 antibody of any one of claims 39 to 51; and (ii) a pharmaceutically acceptable carrier.一種分離的核酸或載體，其編碼如請求項1至38中任一項之多特異性構築體或如請求項39至51中任一項之分離的抗IL-13抗體的一或多個多肽。An isolated nucleic acid or vector encoding one or more polypeptides of the multispecific construct of any one of claims 1 to 38 or the isolated anti-IL-13 antibody of any one of claims 39 to 51.一種宿主細胞，其包含如請求項53之分離的核酸或載體。A host cell comprising the isolated nucleic acid or vector of claim 53.一種治療個體之發炎性疾病的方法，其包含向該個體投予有效量之如請求項52之醫藥組成物。A method for treating an inflammatory disease in a subject, comprising administering to the subject an effective amount of the pharmaceutical composition of claim 52.如請求項55之方法， (i)其中該發炎性疾病為氣喘、異位性皮膚炎或慢性阻塞性肺病(COPD)；及/或 (ii)其中該個體為人類。The method of claim 55,(i) wherein the inflammatory disease is asthma, atopic dermatitis, or chronic obstructive pulmonary disease (COPD); and/or(ii) wherein the individual is a human.一種產生多特異性構築體或抗IL-13抗體構築體之方法，其包含 i)在適合表現該多特異性構築體或該抗IL-13抗體構築體的條件下培養含有如請求項53之分離的核酸或載體的宿主細胞或如請求項54之宿主細胞；及 ii)獲得該表現的多特異性構築體或抗IL-13抗體構築體。A method for producing a multispecific construct or an anti-IL-13 antibody construct, comprising:i) culturing a host cell containing the isolated nucleic acid or vector of claim 53 or the host cell of claim 54 under conditions suitable for expressing the multispecific construct or the anti-IL-13 antibody construct; andii) obtaining the expressed multispecific construct or anti-IL-13 antibody construct.
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