WO2021105669A1

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Publication number: WO2021105669A1
Application number: PCT/GB2020/052998
Authority: WO
Inventors: Daniel John Lightwood; Victoria Louise REDGRAVE-O'DOWD; Kerry Louise Tyson; Simon John DRAPER; Francesca Rose DONNELLAN
Original assignee: Oxford University Innovation Limited
Priority date: 2019-11-29
Filing date: 2020-11-25
Publication date: 2021-06-03
Also published as: GB201917480D0

Abstract

The invention relates to antibodies that bind to the Ebola virus glycoprotein, particularly to antibody cocktails comprising such antibodies. The antibodies and cocktails may be used to treat, prevent or ameliorate Ebola virus infection.

Description

ANTIBODIES

Field of the Invention

The present invention relates to antibodies binding to the Ebola virus glycoprotein. The present invention also relates to pharmaceutical compositions comprising the antibodies and methods of preventing, ameliorating or treating an Ebola virus infection using such antibodies.

Background of the Invention The Zaire Ebola virus (EBOV) outbreak in 2013-2016 in West Africa resulted in

28,616 cases and 11,310 deaths as of June 10, 2016 after which the end of EBOV transmission was declared by the WHO. A new outbreak is in progress in the Democratic Republic of Congo (DRC), which has claimed 2150 lives to date (World Health Organisation, Ebola Virus Disease Democratic Republic of Congo External Situation Report 63, 15th October 2019).

Mixtures of monoclonal antibodies to the EBOV glycoprotein (GP) from convalescent humans (Maruyama et al. 1999, J Virol, 73: 6024-30; Flyak et al. 2016, Cell, 164: 392- 405; Corti et al. 2016, Science, 351: 1339-42; Bornholdt et al. 2016, Science, 351: 1078- 83; Wee et al. 2017, Cell, 169: 878-90 el 5; Flyak et al. 2018, Nat Microbiol, 3: 670-77; Gilchuk et al. 2018, Immunity, 49, 363-374), humanised mice (Pascal et al. 2018, J Infect Dis, 1-15), hyper-immunised macaques (Keck et al. 2016 J Virol, 90: 279-91 ; Zhao et al. 2017 J Biol Chem, 286: 33511-9) and wild-type mice (Furuyama et al. 2016 Sci Rep, 6: 20514; Marzi et al. 2012 PLoS One, 7: e36192; Wilson et al. 2000 Science, 287: 1664-6 ; Qiu et al. 2012 PLoS Negl Trop Dis, 6: el575 ; Pettitt et al. 2013 Sci Transl Med, 5: 199ral3; Takada et al. 2007 Vaccine, 25: 993-9) can be therapeutic in various animal models.

A recent comprehensive study of monoclonal antibodies collected from laboratories across the globe by the Viral Haemorrhagic Fever Immunotherapeutic Consortium (VIC) emphasised the variety of independent epitopes on the viral glycoprotein that can be bound by protective antibodies, and the range of antibody dependent mechanisms that can contribute to protection in vivo (Saphire 2018, Cell, 9, 938-952). The VIC study established that neutralisation in vitro was a strong indicator of the protective potential of an antibody in small animal studies, but in addition revealed that multiple Fc recruited functions also contributed to protection. In these examples the great majority of therapeutic antibodies were isolated from animals or humans after multiple or prolonged exposures to the Ebola glycoprotein.

Three monoclonal antibody therapeutics have recently been evaluated in a clinical trial in humans in the current outbreak in the DRC (NCT03719586): ZMapp, 114 and REGN- 3470-3471-3479 had all previously shown efficacy in non-human primate models as late as five days post infection, and were also compared to a fourth study arm in which patients received Remdesivir. In a recent press release, it was announced that the trial was to be terminated early after an interim analysis of 499 patients with an extension phase continuing in which all future patients be randomized to receive either REGN-3470-3471- 3479 or mAbl 14 (NIAID press release, 12th August 2019, Accessed on 23rd October 2019, via: https://www.niaid.nih.gov/news-events/independent-monitoring-board- recommends-early-termination-ebola-therapeutics-trial-drc). This was due to the reduction in case fatality rates seen in patients in these groups: case fatality rate in the ZMapp arm was 49%, whereas for patients receiving REGN-3470-3471-3479 and 114 the case fatality rates were 29% and 34% respectively. In addition, it was been reported that in these two groups, those patients who were treated soon after infection had a survival rate of 90% (Maxmen, 2019, Nature News doi: 10.1038/d41586-019-02442-6).

ZMapp is a cocktail of murine chimeric antibodies (cl3C6, c2G4 and c4G7), one targeting the glycan cap and two to the base of the GP (Qiu et al. 2014, Nature, 514, 47- 53). An antibody of human origin, 114 (to the receptor binding region) was assessed as a monotherapy (Corti et al. 2016, Science, 351: 1339-42). REGN-3470-3471-3479 is an antibody cocktail developed by Regeneron, derived from humanised mice, containing one antibody to the fusion loop, one to the head, and one glycan cap (Pascal et al. 2018, J Infect Dis, 1-15). This cocktail was intentionally chosen to combine antibodies to independent epitopes with neutralisation and immune effector functions, thought to be complementary. However, there are multiple species of Ebola virus that cause fatal haemorrhagic ebola virus disease (EVD) in humans: alongside EBOV, Sudan Ebola virus (SUDV) and Bundibugyo Ebola virus (BDBV) have caused several outbreaks. In addition Tai Forest Ebola virus (TAFV) caused a single case of non-lethal EVD (Burk et al, FEMS, 2019), and

Reston Ebola virus infections in humans were identified by seroconversion, but did not cause disease (Miranda et al, Lancet, 1991). The recent discovery of a sixth Ebola virus species in bats that can infect human cells in vitro (Goldstein et al, Nat. Microbiol., 2018) highlights the continued threat of the emergence of further Ebola viruses with the potential for spill over into humans. A key limitation of the therapies showing efficacy in the current outbreak is that they are specific to EBOV only.

There is therefore a need for the development of optimised cocktails of antibodies for use in human disease caused by the complete range of Ebola virus species.

More recently there have been several published antibodies that are able to bind GP from multiple Ebola virus species and that are protective in animal models of infection against multiple species of Ebola virus.

Many of these antibodies target the fusion loop of the GP: 6D6 contacts the FL tip (Furuyama et al, Sci Rep, 2016; Milligan et al, J Infect Dis, 2018), CA45 (Zhao et al, Cell, 2017; Janus et al, Nature Comm, 2018; Brannan et al, Nature Comm, 2019) binds along the length of the fusion loop paddle, ADI- 15787 binds across the fusion loop and HR1 region (Wee et al, Cell, 2017; Murin et al, Cell Rep, 2018; West et al, MBio 2018; Wee et al, Cell Host & Microbe, 2019). Additional broadly neutralising or protective mAbs bind non-fusion loop epitopes: ADI-15946 and its matured variant ADI-23774 bind the base of the GP chalice (Wee et al, Cell Host & Microbe 2019; West et al, Nature Structural & Molecular Biology, 2019), and FVM04 which binds the crest of the receptor binding region (Howell et al, Cell Rep, 2016).

As with the REGN cocktail designed to target EBOV, a rationally designed cocktail of therapeutic antibodies would target multiple non-overlapping conserved epitopes affecting multiple functions of the GP. There is therefore rationale to discover protective antibodies that can bind a broad range of Ebola viruses that target epitopes additional to those identified by published antibodies to the fusion loop.

Cocktails of antibodies can show greater therapeutic efficacy than monotherapies by both improving overall protection and reducing severity of symptoms (Qiu et al, Nature, 2014; Howell et al, Cell Rep, 2016; Keck et al, J Virol, 2016; Wee et al, Cell Host & Microbe, 2019; Brannan et al, Nature Comm, 2019; Bornholdt et al, Cell Host & Microbe, 2019); in some cases with synergy between non-neutralising and neutralising antibodies (Howell et al, Cell Rep, 2017). It has also been argued that targeting multiple epitopes using a cocktail of antibodies reduces the chance of viral escape: cocktails of antibodies can reduce transient viremia in comparison to a monotherapy even if both are protective (Corti et al, Science, 2016) and hence reduce opportunities for the virus to evolve to avoid therapeutic antibodies (Saphire and Aman, Trends in Microbiol., 2016). Conserved epitopes such as those targeted by pan-Ebola virus antibodies may also incur a higher fitness cost to viral escape mutations, decreasing the chances of viral escape from antibodies targeting these regions. However, single point mutations have been identified that abolish binding by broadly-protective antibodies without affecting viral entry into cells (Wee et al, Cell, 2017), highlighting the need for antibodies to be used in combination therapies.

Summary of the Invention

The present invention provides an anti-Ebola virus antibody that recognises an epitope on the Ebola virus glycoprotein comprising one or more residues selected from the group consisting of F132, P133, R134, C135, R136, Y137, V138, H139, K140, V141, S142 and G143 of SEQ ID NO: 22.

In addition, the invention provides an antibody that binds to the same epitope as, or competes for binding with, an antibody of the invention.

The present invention also provides a nucleic acid or a pair of nucleic acids, optionally mRNA, encoding the heavy and light chains of an antibody of the invention, an expression vector comprising the nucleic acid(s) or a pair of expression vectors comprising the pair of nucleic acids, a host cell comprising the expression vector(s) and a method of producing an antibody of the invention, said method comprising culturing the host cell under conditions permitting production of the antibody and recovering the antibody so produced.

Furthermore, the invention provides a pharmaceutical composition comprising an antibody of the invention, or a nucleic acid/pair of nucleic acids of the invention.

The invention also provides a method of treating, preventing or ameliorating Ebola virus infection, the method comprising administering a composition of the invention to a subject in need thereof. Likewise, the invention provides a composition of the invention for use in a method of treating, preventing or ameliorating Ebola virus infection, the method comprising administering the composition to a subject in need thereof and use of a composition of the invention in the manufacture of a medicament for treating, preventing or ameliorating Ebola virus infection. Brief Description of the Figures

Figure 1 shows binding of broadly reactive monoclonal antibodies to HEK293 cells transiently transfected with GP from different Ebola virus species or irrelevant antigen (mock). Mean and range of duplicates shown. Binding confirmed in two other experiments (TAFV GP binding for mabs 11886, 11881, 11889, 11892 confirmed in i other experiment only).

Figure 2 shows a table of inhibitory concentration at 50% (IC50) and 90% (IC90) values for in vitro neutralisation of S-FLU viruses pseudotyped with GP from EBOV, SUDV and BDBV. Strong: neutralises >90% of virus at tested concentrations; neut: neutralises virus to 90% at tested concentrations; partial: cannot neutralise 90% of virus at tested concentrations; nn: non-neutralising. *reached 90% at highest concentration tested. CA45 is a published broadly neutralising monolconal antibody (Zhao et al, cell, 2017).

Figure 3 shows in vitro neutralisation of Ebola virus GP pseudotyped S-FLU viruses by broadly-reactive monoclonal antibodies. Mean and range of duplicates shown for each assay. Results confirmed in independent repeat of experiment.

Figure 4 shows that broadly-reactive rabbit antibodies 11897, 11883 and 11889 compete for binding to EBOV GP with monoclonal antibodies with known epitopes in the glycan cap region in a competitive immunofluorescence assay. 21-D8-5A: influenza neuraminidase-specific antibody acting as a negative control; 6541, 66-4-cl2, c2G4 and c4G7: overlapping base epitopes; 6660 and 6662: receptor binding region epitopes; 66-3- 9C, 66-3-2C, 040 and cl3C6: different glycan cap epitopes; CA45, 6D6, FVM02, and ADI-15878 bind to the conserved fusion loop. Self indicates signal when competed with unbiotinylated version of the same monoclonal antibody. Error bars show mean and 95% confidence intervals for six replicates within assay.

Figure 5 shows that broadly-reactive rabbit antibodies 11886 and 11892 compete for binding to EBOV GP with monoclonal antibodies with known epitopes in the base region in a competitive immunofluorescence assay. 21-D8-5A: influenza neuraminidase-specific antibody acting as a negative control; 6541, 66-4-cl2, c2G4 and c4G7: overlapping base epitopes; 6660 and 6662: receptor binding region epitopes; 66-3-9C, 66-3-2C, 040 and cl3C6: different glycan cap epitopes; CA45, 6D6, FVM02, and ADI-15878 bind to the conserved fusion loop. Self indicates signal when competed with unbiotinylated version of the same monoclonal antibody. Error bars show mean and 95% confidence intervals for six replicates within assay.

Figure 6 shows binding of broadly reactive rabbit antibodies to EBOV GP and SUDV GP digested with thermolysin. Closed points: thermolysin treated GP; open points: untreated GP. Error bars show mean and range of 6 replicates per thermolysin treated sample and 3 replicates per undigested sample. 040: glycan cap antibody; 6541: base binding antibody; MR78: receptor binding region antibody that can only bind GP in the absence of the glycan cap.

Figure 7 shows logo plots generated by MEME tool from top 100 enriched sequences from panning 9mer and 13mer peptide phage display libraries with antibody 11886.

Motifs with an e value <0.05 only were considered significant.

Figure 8 shows alignment of GP sequences with meme-derived motifs from peptide sequences enriched across three experiments and two peptide phage libraries by antibody 11886.

Figure 9 shows arginine residues in 11886 binding peptide motifs highlighted on existing electron microscopy structure of EBOV GP. Key arginine residues highlighted in stick representation. Cartoon representation of PDB structure 5KEL (Pallesen et al, Nature Microbiology, 2016).

Figure 10 shows 11886 binding peptide motifs highlighted on existing electron microscopy structures of EBOV GP in complex with published fabs. The 11886 binding motif derived from peptide phage display is shown in rectangles. A. Cartoon representation of PDB structure 5KEL (Pallesen et al, Nature Microbiology, 2016). B. Surface rendering of 5KEL generated in Pymol with cl3C6 fab and c2G4 fab. C. Cartoon representation of PDB structure 6MAM (West et al, Nature Structural and Molecular Methods, 2019). D. Surface rendering of 6MAM generated in Pymol with ADI-15946 fab: teal. E. Surface rendering of 5KEL generated in Pymol with fabs hidden. FVM04 and m2 ID 10 contacts are shown.

Brief Description of the Sequence Listing

SEQ ID NO: 1 shows the 11886 CDRL1 SEQ ID NO: 2 shows the 11886 CDRL2 SEQ ID NO: 3 shows the 11886 CDRL3 SEQ ID NO: 4 shows a 11886 CDRL3 variant

SEQ ID NO: 5 shows a 11886 CDRL3 variant

SEQ ID NO: 6 shows the 11886 CDRH1

SEQ ID NO: 7 shows the 11886 CDRH2

SEQ ID NO: 8 shows the 11886 CDRH3

SEQ ID NO: 9 shows a 11886 CDRH3 variant

SEQ ID NO: 10 shows a 11886 CDRH3 variant

SEQ ID NO: 11 shows the rabbit 11886 VL (amino acid)

SEQ ID NO: 12 shows the rabbit 11886 VL (nucleic acid)

SEQ ID NO: 13 shows the rabbit 11886 VH (amino acid)

SEQ ID NO: 14 shows the rabbit 11886 VH (nucleic acid)

SEQ ID NO: 15 shows a humanised 11886 VL

SEQ ID NO: 16 shows a humanised 11886 VL (with CDRL3 variant)

SEQ ID NO: 17 shows a humanised 11886 VL (with CDRL3 variant)

SEQ ID NO: 18 shows a humanised 11886 VH

SEQ ID NO: 19 shows a humanised 11886 VH (with CDRH3 variant)

SEQ ID NO: 20 shows a humanised 11886 VH (with CDRH3 variant)

SEQ ID NO: 21 shows the Bundibugyo Ebola virus glycoprotein sequence (NBCI reference YP_003815435.1).

SEQ ID NO: 22 shows the Zaire Ebola virus glycoprotein sequence (ATY51135.1) SEQ ID NO: 23 shows the Sudan Ebola virus glycoprotein sequence (NCBI Reference Sequence: YP 138523.1)

SEQ ID NO: 24 shows the Tai Forest Ebola virus glycoprotein sequence (NCBI Reference Sequence: YP 003815426.1)

SEQ ID NO: 25 shows the Ebola virus glycoprotein consensus motif SEQ ID NOs: 26-33 show 9 and 13 mer peptide motifs which mapped onto Ebola glycoprotein sequences

SEQ ID NO: 34 shows the 11897 CDRLl SEQ ID NO: 35 shows the 11897 CDRL2 SEQ ID NO: 36 shows the 11897 CDRL3 SEQ ID NO: 37 shows the 11897 CDRHl SEQ ID NO: 38 shows the 11897 CDRH2 SEQ ID NO: 39 shows a 11897 CDRH2 variant SEQ ID NO: 40 shows a 11897 CDRH2 variant SEQ ID NO: 41 shows a 11897 CDRH2 variant SEQ ID NO: 42 shows the 11897 CDRH3 SEQ ID NO: 43 shows the 11897 rabbit VL (amino acid)

SEQ ID NO: 44 shows the 11897 rabbit VL (nucleic acid)

SEQ ID NO: 45 shows the 11897 rabbit VH (amino acid)

SEQ ID NO: 46 shows the 11897 rabbit VH (nucleic acid)

SEQ ID NO: 47 shows a humanised 11897 VL

SEQ ID NO: 48 shows a humanised 11897 VH

SEQ ID NO: 49 shows a humanised 11897 VH (CDRH2 variant)

SEQ ID NO: 50 shows a humanised 11897 VH (CDRH2 variant)

SEQ ID NO: 51 shows a humanised 11897 VH (CDRH2 variant)

SEQ ID NO: 52 shows the 11878 CDRL1

SEQ ID NO: 53 shows the 11878 CDRL2

SEQ ID NO: 54 shows the 11878 CDRL3

SEQ ID NO: 55 shows the 11878 CDRH1

SEQ ID NO: 56 shows the 11878 CDRH2

SEQ ID NO: 57 shows the 11878 CDRH3

SEQ ID NO: 58 shows the 11878 rabbit VL (amino acid)

SEQ ID NO: 59 shows the 11878 rabbit VL (nucleic acid)

SEQ ID NO: 60 shows the 11878 rabbit VH (amino acid)

SEQ ID NO: 61 shows the 11878 rabbit VH (nucleic acid)

SEQ ID NO: 62 shows a humanised 11878 VL SEQ ID NO: 63 shows a humanised 11878 VH SEQ ID NO: 64 shows the 11883 CDRL1 SEQ ID NO: 65 shows the 11883 CDRL2 SEQ ID NO: 66 shows the 11883 CDRL3 SEQ ID NO: 67 shows the 11883 CDRH1 SEQ ID NO: 68 shows a 11883 CDRH1 variant SEQ ID NO: 69 shows a 11883 CDRH1 variant

SEQ ID NO: 70 shows the 11883 CDRH2 SEQ ID NO: 71 shows a 11883 CDRH2 variant SEQ ID NO: 72 shows a 11883 CDRH2 variant SEQ ID NO: 73 shows the 11883 CDRH3 SEQ ID NO: 74 shows the 11883 rabbit VL (amino acid)

SEQ ID NO: 75 shows the 11883 rabbit VL (nucleic acid)

SEQ ID NO: 76 shows the 11883 rabbit VH (amino acid)

SEQ ID NO: 77 shows the 11883 rabbit VH (nucleic acid)

SEQ ID NO: 78 shows a humanised 11883 VL

SEQ ID NO: 79 shows a humanised 11883 VH

SEQ ID NO: 80 shows a humanised 11883 VH (CDRH1 and 2 variant)

SEQ ID NO: 81 shows a humanised 11883 VH (CDRH1 and 2 variant)

SEQ ID NO: 82 shows a humanised 11883 VH (CDRH1 and 2 variant)

SEQ ID NO: 83 shows the 11889 CDRL1

SEQ ID NO: 84 shows the 11889 CDRL2

SEQ ID NO: 85 shows the 11889 CDRL3

SEQ ID NO: 86 shows the 11889 CDRH1

SEQ ID NO: 87 shows a 11889 CDRH1 variant

SEQ ID NO: 88 shows the 11889 CDRH2

SEQ ID NO: 89 shows a 11889 CDRH2 variant

SEQ ID NO: 90 shows the 11889 CDRH3

SEQ ID NO: 91 shows the 11889 rabbit VL (amino acid)

SEQ ID NO: 92 shows the 11889 rabbit VL (nucleic acid)

SEQ ID NO: 93 shows the 11889 rabbit VH (amino acid)

SEQ ID NO: 94 shows the 11889 rabbit VH (nucleic acid)

SEQ ID NO: 95 shows a humanised 11889 VL SEQ ID NO: 96 shows a humanised 11889 VH

SEQ ID NO: 97 shows a humanised 11889 VH (CDRH1 and 2 variant)

SEQ ID NO: 98 shows the 11892 CDRL1

SEQ ID NO: 99 shows the 11892 CDRL2

SEQ ID NO: 100 shows the 11892 CDRL3

SEQ ID NO: 101 shows a 11892 CDRL3 variant

SEQ ID NO: 172 shows a 6662 variant CDRH2 SEQ ID NO: 173 shows a 6662 variant CDRH2 SEQ ID NO: 174 shows a 6662 variant CDRH3 SEQ ID NO: 175 shows a 6662 variant CDRH3 SEQ ID NO: 176 shows a 6662 variant CDRL3

SEQ ID NO: 177 shows a 6662 variant CDRL3 SEQ ID NO: 178 shows a 6662 variant CDRL3 Detailed Description of the Invention

It is to be understood that different applications of the disclosed products and methods may be tailored to the specific needs in the art. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments of the invention only, and is not intended to be limiting. In addition as used in this specification and the appended claims, the singular forms

“a”, “an”, and “the” include plural referents unless the content clearly dictates otherwise. Thus, for example, reference to “an amino acid sequence” includes two or more such sequences, and the like.

All publications, patents and patent applications cited herein, whether supra or infra, are hereby incorporated by reference in their entirety.

Anti-Ebola virus antibodies

The present invention relates to antibodies that bind to (recognise) the Ebola virus glycoprotein (GP) and to pharmaceutical compositions comprising such antibodies. There are currently six Ebola virus species (Zaire, Sudan, Bundibugyo, Reston, Tai Forest and Bombali) and many different strains. Zaire, Sudan, Bundibugyo and Tai Forest cause disease in humans, with Zaire being the most deadly. The Ebola vims glycoprotein is the only virally expressed protein on the vims surface and is critical for attachment to host cells and catalysis of membrane fusion. The glycoprotein is cleaved by furin to form a disulphide-linked GP1-GP2 heterodimer, which assembles as trimers on the vims surface. GP1 contains the receptor-binding site responsible for host cell attachment, the glycan cap and the mucin-like domain, whereas GP2 contains heptad repeats and a transmembrane domain.

Ebola vims glycoprotein sequences vary between species. The nucleotide and amino acid sequences of the glycoprotein from various Ebola vims species/strains have been determined, with examples including GenBank: AF086833.2 providing the complete genome of Zaire Mayinga, and GenBank: AAN37507.1, GenBank: AAG40168.1 and GenBank: ATY51135.1 providing examples of Zaire glycoprotein sequences. Accession numbers NC_014373.1 and NC_006432.1 provide the complete genome sequences of Budibugyo and Sudan Gulu. Accession numbers GenBank: AGL73446.1, GenBank: AGL73439.1 and NCBI Reference Sequence: YP 138523.1 provide examples of a Sudan glycoprotein sequence and accession numbers GenBank: AGL73474.1, GenBank: AGL73467.1 and NCBI Reference Sequence: YP 003815435.1 provide examples of Bundibugyo glycoprotein sequences. Accession number NCBI Reference Sequence:

YP 003815426.1 provides a Tai Forest glycoprotein sequence. Other sequences are readily available from sequence databases, such as GenBank and UniProt.

Antibodies of the invention may be “isolated” antibodies. An isolated antibody is an antibody which is substantially free of other antibodies having different antigenic specificities.

The term “antibody” as referred to herein includes whole antibodies and any antigen binding fragment (i.e., “antigen-binding portion”) or single chains thereof. An antibody refers to a glycoprotein comprising at least two heavy (H) chains and two light (L) chains inter-connected by disulfide bonds, or an antigen-binding portion thereof. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. Each light chain is comprised of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The variable regions of the heavy and light chains contain a binding domain that interacts with an antigen. The VH and VL regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR).

The constant regions of the antibodies may mediate the binding of the immunoglobulin to host tissues or factors, including various cells of the immune system (e.g., effector cells) and the first component (Clq) of the classical complement system.

Antibodies of the invention are typically monoclonal antibodies. An antibody of the invention may be a chimeric antibody, a CDR-grafted antibody, a nanobody, a human or humanised antibody or an antigen-binding portion of any thereof. Typically, the antibody is a humanised antibody. Fully human antibodies are those antibodies in which the variable regions and the constant regions (where present) of both the heavy and the light chains are all of human origin, or substantially identical to sequences of human origin, but not necessarily from the same antibody.

The antibody molecules of the present invention may comprise a complete antibody molecule having full length heavy and light chains or a fragment or antigen-binding portion thereof. The term "antigen-binding portion" of an antibody refers to one or more fragments of an antibody that retain the ability to selectively bind to an antigen. It has been shown that the antigen-binding function of an antibody can be performed by fragments of a full-length antibody. The antibodies and fragments and antigen binding portions thereof may be, but are not limited to Fab, modified Fab, Fab’, modified Fab’, F(ab’)2, Fv, single domain antibodies (e.g. VH or VL or VHH), scFv, bi, tri or tetra-valent antibodies, Bis-scFv, diabodies, triabodies, tetrabodies and epitope-binding fragments of any of the above (see for example Holliger and Hudson, 2005, Nature Biotech. 23(9): 1126- 1136; Adair and Lawson, 2005, Drug Design Reviews - Online 2(3), 209-217). The methods for creating and manufacturing these antibody fragments are well known in the art (see for example Verma et ak, 1998, Journal of Immunological Methods, 216, 165-181). Other antibody fragments for use in the present invention include the Fab and Fab’ fragments described in International patent applications WO 2005/003169, WO 2005/003170 and WO 2005/003171 and Fab-dAb fragments described in International patent application W02009/040562. Multi-valent antibodies may comprise multiple specificities or may be monospecific (see for example WO 92/22853 and WO 05/113605 and the DVD-Ig as disclosed in WO 2007/024715, or the so-called (FabFv)2Fc described in WO 2011/030107). An alternative multi-specific antigen-binding fragment comprises a Fab linked to two scFvs or dsscFvs, each scFv or dsscFv binding the same or a different target (e.g., one scFv or dsscFv binding a therapeutic target and one scFv or dsscFv that increases half-life by binding, for instance, albumin). Such antibody fragments are described in International Patent Application Publication No, WO 2015/197772.

These antibody fragments may be obtained using conventional techniques known to those of skill in the art, and the fragments may be screened for utility in the same manner as intact antibodies.

The constant region domains of the antibody molecule of the present invention, if present, may be selected having regard to the proposed function of the antibody molecule, and in particular the effector functions which may be required. For example, the constant region domains may be IgA, IgD, IgE, IgG or IgM domains, typically IgG (i.e. IgGl,

IgG2, IgG3 or IgG4). Typically, the constant regions are human. In particular, human IgG (i.e. IgGl, IgG2, IgG3 or IgG4) constant region domains may be used. Typically, a human IgGl constant region.

The light chain constant region may be either lambda or kappa.

Antibodies of the invention may be mono-specific or multi-specific (e.g. bi-specific).

A multi-specific antibody comprises at least two different variable domains, wherein each variable domain is capable of binding to a separate antigen or to a different epitope on the same antigen.

An antibody of the invention may be a human antibody. The term "human antibody", as used herein, is intended to include antibodies having variable regions in which both the framework and CDR regions are derived from human germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, the constant region also is derived from human germline immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo). However, the term "human antibody", as used herein, is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, have been grafted onto human framework sequences.

The term “human antibody derivatives” refers to any modified form of the human antibody, e.g., a conjugate of the antibody and another agent or antibody. It will also be understood by one skilled in the art that antibodies may undergo a variety of posttranslational modifications. The type and extent of these modifications often depends on the host cell line used to express the antibody as well as the culture conditions. Such modifications may include variations in glycosylation, methionine oxidation, diketopiperazine formation, aspartate isomerization and asparagine deamidation. A frequent modification is the loss of a carboxy-terminal basic residue (such as lysine or arginine) due to the action of carboxypeptidases (as described in Harris, RJ. Journal of Chromatography 705:129-134, 1995).

Biological molecules, such as antibodies or fragments, contain acidic and/or basic functional groups, thereby giving the molecule a net positive or negative charge. The amount of overall “observed” charge will depend on the absolute amino acid sequence of the entity, the local environment of the charged groups in the 3D structure and the environmental conditions of the molecule. The isoelectric point (pi) is the pH at which a particular molecule or surface carries no net electrical charge. In one embodiment the antibody or fragment according to the present disclosure has an isoelectric point (pi) of at least 7. In one embodiment the antibody or fragment has an isoelectric point of at least 8, such as 8.5, 8.6, 8.7, 8.8 or 9. In one embodiment the pi of the antibody is 8. Programs such as ** ExPASY http://www.expasy.ch/tools/pi_tool.html (see Walker, The Proteomics Protocols Handbook, Humana Press (2005), 571-607), may be used to predict the isoelectric point of the antibody or fragment.

Antibodies may be obtained by administering polypeptides to an animal, e.g. a non human animal, using well-known and routine protocols, see for example Handbook of Experimental Immunology, D. M. Weir (ed.), Vol 4, Blackwell Scientific Publishers, Oxford, England, 1986). Many warm-blooded animals, such as rabbits, mice, rats, sheep, cows, camels or pigs may be immunized. However, mice, rabbits, pigs and rats are generally most suitable.

Monoclonal antibodies may be prepared by any method known in the art such as the hybridoma technique (Kohler & Milstein, 1975, Nature, 256:495-497), the trioma technique, the human B-cell hybridoma technique (Kozbor et ah, 1983, Immunology Today, 4:72) and the EBV-hybridoma technique (Cole et ah, Monoclonal Antibodies and Cancer Therapy, pp77-96, Alan R Liss, Inc., 1985). Antibodies may also be generated using single lymphocyte antibody methods by cloning and expressing immunoglobulin variable region cDNAs generated from single lymphocytes selected for the production of specific antibodies by for example the methods described by Babcook, J. et al., 1996, Proc. Natl. Acad. Sci. USA 93(15): 7843-78481; WO92/02551; W02004/051268 and W02004/106377.

The antibodies can also be generated using various phage display methods known in the art and include those disclosed by Brinkman et al. (in J. Immunol. Methods, 1995, 182: 41-50), Ames et al. (J. Immunol. Methods, 1995, 184:177-186), Kettleborough et al. (Eur. J. Immunol. 1994, 24:952-958), Persic et al. (Gene, 1997 1879-18), Burton et al. (Advances in Immunology, 1994, 57:191-280) and WO 90/02809; WO 91/10737; WO 92/01047; WO 92/18619; WO 93/11236; WO 95/15982; WO 95/20401; and US

5,698,426; 5,223,409; 5,403,484; 5,580,717; 5,427,908; 5,750,753; 5,821,047; 5,571,698; 5,427,908; 5,516,637; 5,780,225; 5,658,727; 5,733,743 and 5,969,108.

Fully human antibodies are those antibodies in which the variable regions and the constant regions (where present) of both the heavy and the light chains are all of human origin, or substantially identical to sequences of human origin, but not necessarily from the same antibody. Examples of fully human antibodies may include antibodies produced, for example by the phage display methods described above and antibodies produced by mice in which the murine immunoglobulin variable and optionally the constant region genes have been replaced by their human counterparts e.g. as described in general terms in EP

0546073, US 5,545,806, US 5,569,825, US 5,625,126, US 5,633,425, US 5,661,016, US 5,770,429, EP 0438474 and EP 0463151.

The term “humanized antibody” is intended to refer to CDR-grafted antibody molecules in which CDR sequences derived from the germline of another mammalian species, such as a mouse or rabbit, have been grafted onto human framework sequences. Additional framework region modifications may be made within the human framework sequences and also within the CDR sequences.

As used herein, the term ‘CDR-grafted antibody molecule’ refers to an antibody molecule wherein the heavy and/or light chain contains one or more CDRs (including, if desired, one or more modified CDRs) from a donor antibody (e.g. a murine, rat or rabbit monoclonal antibody) grafted into a heavy and/or light chain variable region framework of an acceptor antibody (e.g. a human antibody). For a review, see Vaughan et al, Nature Biotechnology, 16, 535-539, 1998. In one embodiment rather than the entire CDR being transferred, only one or more of the specificity determining residues from any one of the CDRs described herein are transferred to the human antibody framework (see for example, Kashmiri et al., 2005, Methods, 36, 25-34). In one embodiment only the specificity determining residues from one or more of the CDRs described herein are transferred to the human antibody framework. In another embodiment only the specificity determining residues from each of the CDRs described herein are transferred to the human antibody framework.

When the CDRs or specificity determining residues are grafted, any appropriate acceptor variable region framework sequence may be used having regard to the class/type of the donor antibody from which the CDRs are derived, including mouse, primate and human framework regions. Suitably, the CDR-grafted antibody according to the present invention has a variable domain comprising human acceptor framework regions as well as one or more of the CDRs or specificity determining residues described above. Thus, provided in one embodiment is a CDR-grafted antibody wherein the variable domain comprises human acceptor framework regions and non-human donor CDRs.

Examples of human frameworks which can be used are KOL, NEWM, REI, EU, TUR, TEI, LAY and POM (Rabat et al., supra). For example, KOL and NEWM can be used for the heavy chain, REI can be used for the light chain and EU, LAY and POM can be used for both the heavy chain and the light chain. Alternatively, human germline sequences may be used; these are available for example at: http://www.vbase2.org/ (see Retter et al, Nucl. Acids Res. (2005) 33 (supplement 1), D671-D674).

In a CDR-grafted antibody, the acceptor heavy and light chains do not necessarily need to be derived from the same antibody and may, if desired, comprise composite chains having framework regions derived from different chains.

Also, in a CDR-grafted antibody, the framework regions need not have exactly the same sequence as those of the acceptor antibody. For instance, unusual residues may be changed to more frequently occurring residues for that acceptor chain class or type. Alternatively, selected residues in the acceptor framework regions may be changed so that they correspond to the residue found at the same position in the donor antibody (see Reichmann et al., 1998, Nature, 332, 323-324). Such changes should be kept to the minimum necessary to recover the affinity of the donor antibody. A protocol for selecting residues in the acceptor framework regions which may need to be changed is set forth in WO 91/09967.

In one aspect, the invention provides an anti-Ebola virus antibody that recognises (binds to) an epitope on the Ebola glycoprotein comprising one or more residues selected from the group consisting of F132, P133, R134, C135, R136, Y137, V138, H139, K140, V141, S142 and G143 of SEQ ID NO: 22. SEQ ID NO: 22 is the Zaire Ebola virus glycoprotein sequence with accession number ATY51135.1. Residues F132, P133, R134, C135, R136, Y137, V138, H139, K140 V141, S142 and G143 are conserved in Budibugyo (NBCI reference YP 003815435.1; SEQ ID NO: 21) and Tai forest (NCBI Reference Sequence: YP 003815426.1; SEQ ID NO: 24). In Sudan (NCBI Reference Sequence:

YP 138523.1; SEQ ID NO: 23) residue 141 is A and residue 142 is Q (see Figure 8).

Antibodies of the invention also therefore typically recognise an epitope comprising

(i) one or more residues selected from the group consisting of F132, P133, R134, C135, R136, Y137, V138, H139, K140, V141, S142 and G143 of SEQ ID NO: 21 or 24 and/or

(ii) one or more residues selected from the group consisting of F132, P133, R134, C135, R136, Y137, V138, H139, K140, A141, Q142 and G143 of SEQ ID NO: 23. In particular, antibodies of the invention recognise an epitope comprising one or more residues selected from the group consisting of F132, P133, R134, C135, R136, Y137, V138, H139 and K140 of any one of SEQ ID NOs: 21-24, preferably in all of SEQ ID NOs: 21-24.

Typically, antibodies of the invention recognise an epitope comprising one or more residues selected from the group consisting of R134, C135 and R136 (of any one of, preferably all of, SEQ ID NOs: 21-24. In some instances antibodies of the invention recognise an epitope comprising R134 and R136, or all three of R134, C135 and R136.

In some instances, the epitope may further comprise one of more residues selected from group consisting of G102, E103, W104, A105, E106, N107 and C108 and/or (b) one of more residues selected from group consisting of R605, W606, G607, G608, T609, C610 and H611 . The residue numbering may be according to SEQ ID NO: 22. However, these residues are conserved in Zaire, Bundibugyo, Sudan and Tai Forest. Therefore the residue numbering may also be according to SEQ ID NO: 21, 23 or 24.

As these antibodies recognise epitopes which are conserved across species, the antibodies are cross-reactive for all of Zaire, Bundibugyo, Sudan and Tai Forest To screen for antibodies that bind to a particular epitope, a routine cross-blocking assay such as that described in Antibodies, Harlow and Lane (Cold Spring Harbor Press, Cold Spring Harb., NY) can be performed. Other methods include alanine scanning mutants, peptide blots (Reineke (2004) Methods Mol Biol 248:443-63), or peptide cleavage analysis. In addition, methods such as hydrogen deuterium exchange, epitope excision, epitope extraction and chemical modification of antigens can be employed (Tomer (2000) Protein Science 9: 487-496). Such methods are well known in the art.

The antibody epitope may also be determined by electron microscopy.

In addition, antibody epitopes be determined by x-ray crystallography analysis. Antibodies of the present invention may therefore be assessed through x-ray crystallogray analysis of the antibody bound to the Ebola virus glycorptein. Epitopes may, in particular, be identified in this way by determining residues within 4Ά (possibly 5 A) of an antibody paratope residue.

In the examples below, the epitope residues of the 11886 antibody were determined by screening peptide libraries and mapping the peptides bound by the antibody onto the Ebola virus sequences. Such methods (e.g. peptide scanning) may be used in identifying antibodies binding to the regions specified in the claims. For example, an antibody of the invention may bind to peptides comprising the motifs as shown in SEQ ID NOs: 26-33. Peptides may also comprise residue numbers 102-108, 132-143 or 605-611 of any one of SEQ ID NOs: 21-24.

An antibody of the invention may in particular comprise one or more CDR sequences selected from the following:

(a) a CDRLl sequence of SEQ ID NO: 1;

(b)a CDRL2 sequence of SEQ ID NO: 2;

(c)a CDRL3 sequence of SEQ ID NO: 3, 4 or 5;

(d)a CDRHl sequence of SEQ ID NO: 6;

(e)a CDRH2 sequence of SEQ ID NO: 7; and

(f) a CDRH3 sequence of SEQ ID NO: 8, 9 and 10.

SEQ ID NOs: 1, 2, 3, 6, 7 and 8 are the parental rabbit antibody CDR sequences of 11886. SEQ ID NOs: 4, 5, 9 and 10 are humanized variants of the rabbit CDRs.

For example, an antibody of the invention may comprise at least one, at least two or all three heavy chain CDR sequences of SEQ ID NOs: 6, 7 and 8/9/10. An antibody of the invention may also comprise at least one, at least two or all three light chain CDR sequences of SEQ ID NOs: 1, 2 and 3/4/5.

Typically, the antibody of the invention comprises at least one heavy chain CDR sequence selected from SEQ ID NOs: 6-10 and at least one light chain CDR sequence selected from SEQ ID NOSs: 1-5. The antibody of the invention may comprise at least two heavy chain CDR sequences selected from SEQ ID NOs: 6-10 and at least two light chain CDR sequences selected from SEQ ID NOs: 1-5. In some instances, an antibody of the invention may comprise HCDR3 of SEQ ID NO: 8, 9 or 10.

An antibody of the invention may in particular comprise the following CDRs:

(a) a CDRLl sequence of SEQ ID NO: 1;

(b) a CDRL2 sequence of SEQ ID NO: 2;

(c) a CDRL3 sequence of SEQ ID NO: 3, 4 or 5;

(d) a CDRH1 sequence of SEQ ID NO: 6;

(e) a CDRH2 sequence of SEQ ID NO: 7; and

(f) a CDRH3 sequence of SEQ ID NO: 8, 9 or 10.

For example, an antibody of the invention may comprise the following:

(a) a CDRLl sequence of SEQ ID NO: 1;

(b) a CDRL2 sequence of SEQ ID NO: 2;

(c) a CDRL3 sequence of SEQ ID NO: 3;

(d) a CDRH1 sequence of SEQ ID NO: 6;

(e) a CDRH2 sequence of SEQ ID NO: 7; and

(f) a CDRH3 sequence of SEQ ID NO : 8.

An antibody of the invention may comprise a heavy chain variable region sequence of SEQ ID NO: 13 (the VH of 11886). An antibody of the invention may comprise a light chain variable region sequence of SEQ ID NO: 11 (the VL of 11886). An antibody of the invention may comprise a VH and VL sequence pair of SEQ ID NOs: 13 and 11.

An antibody of the invention may also comprise a VH of SEQ ID NO: 18, 19 or 20. These are humanised sequences of 11886. An antibody of the invention may comprise a VL of SEQ ID NO: 15, 16 or 17. These are also humanised sequences of 11886. In particular an antibody of the invention may comprise a VH selected from the group consisting of SEQ ID NOs: 18, 19 and 20 and a VL selected from the group consisting of SEQ ID NOs: 15, 16 and 17. An antibody of the invention may also comprise one or more CDR sequences selected from the following:

(a)a CDRL1 sequence of SEQ ID NO: 34;

(b)a CDRL2 sequence of SEQ ID NO: 35;

(c)a CDRL3 sequence of SEQ ID NO: 36;

(d)a CDRH1 sequence of SEQ ID NO: 37;

(e)a CDRH2 sequence of SEQ ID NO: 38, 39, 40 and 41; and

(f) a CDRH3 sequence of SEQ ID NO: 42.

SEQ ID NOs: 34, 35, 36, 37, 38 and 42 are the parental rabbit antibody CDR sequences of 11897. SEQ ID NOs: 39, 40 and 31 are humanized variants of the rabbit CDRs.

For example, an antibody of the invention may comprise at least one, at least two or all three heavy chain CDR sequences of SEQ ID NOS: 37, 38/39/40/41 and 42.

An antibody of the invention may also comprise at least one, at least two or all three light chain CDR sequences of SEQ ID NOS: 34, 35 and 36.

Typically, the antibody of the invention comprises at least one heavy chain CDR sequence selected from SEQ ID NOS: 37-42 and at least one light chain CDR sequence selected from SEQ ID NOS 34-36. The antibody of the invention may comprise at least two heavy chain CDR sequences selected from SEQ ID NOS: 37-42 and at least two light chain CDR sequences selected from SEQ ID NOS: 34-36. In some instances, an antibody of the invention may comprise HCDR3 of SEQ ID NO: 42.

An antibody of the invention may in particular comprise the following CDRs:

(a)a CDRLl sequence of SEQ ID NO: 34;

(b)a CDRL2 sequence of SEQ ID NO: 35;

(c)a CDRL3 sequence of SEQ ID NO: 36;

(d)a CDRH1 sequence of SEQ ID NO: 37;

(e)a CDRH2 sequence of SEQ ID NO: 38, 39, 40 or 41; and

(f) a CDRH3 sequence of SEQ ID NO: 42.

For example, an antibody of the invention may comprise the following:

(a)a CDRLl sequence of SEQ ID NO: 34;

(b)a CDRL2 sequence of SEQ ID NO: 35;

(c)a CDRL3 sequence of SEQ ID NO: 36; (d)a CDRH1 sequence of SEQ ID NO: 37;

(e)a CDRH2 sequence of SEQ ID NO: 38; and

(f) a CDRH3 sequence of SEQ ID NO: 42.

An antibody of the invention may comprise a heavy chain variable region sequence of SEQ ID NO: 45 (the VH of 11897). An antibody of the invention may comprise a light chain variable region sequence of SEQ ID NO: 43 (the VL of 11897). An antibody of the invention may comprise a VH and VL sequence pair of SEQ ID NOs: 45 and 43.

An antibody of the invention may also comprise a VH of SEQ ID NO: 48, 49, 50 or 51. These are humanised sequences of 11897. An antibody of the invention may comprise a VL of SEQ ID NO: 47. This is also a humanised sequence of 11897. In particular an antibody of the invention may comprise a VH selected from the group consisting of SEQ ID NOs: 48, 49, 50 and 51 and a VL of SEQ ID NO: 47.

An antibody of the invention may also comprise one or more CDR sequences selected from the following:

(a) a CDRL1 sequence of SEQ ID NO: 52;

(b)a CDRL2 sequence of SEQ ID NO: 53;

(c)a CDRL3 sequence of SEQ ID NO: 54;

(d)a CDRH1 sequence of SEQ ID NO: 55;

(e)a CDRH2 sequence of SEQ ID NO: 56; and

(f) a CDRH3 sequence of SEQ ID NO: 57.

SEQ ID NOs: 52-57 are the rabbit antibody CDR sequences of 11878.

For example, an antibody of the invention may comprise at least one, at least two or all three heavy chain CDR sequences of SEQ ID NOS: 55, 56 and 57.

An antibody of the invention may also comprise at least one, at least two or all three light chain CDR sequences of SEQ ID NOS: 52, 53 and 54.

Typically, the antibody of the invention comprises at least one heavy chain CDR sequence selected from SEQ ID NOS: 55-57 and at least one light chain CDR sequence selected from SEQ ID NOS 52-54. The antibody of the invention may comprise at least two heavy chain CDR sequences selected from SEQ ID NOS: 55-57 and at least two light chain CDR sequences selected from SEQ ID NOS: 52-54. In some instances, an antibody of the invention may comprise a HCDR3 of SEQ ID NO: 57.

An antibody of the invention may in particular comprise the following CDRs: (a) a CDRL1 sequence of SEQ ID NO: 52;

(b) a CDRL2 sequence of SEQ ID NO: 53;

(c) a CDRL3 sequence of SEQ ID NO: 54;

(d) a CDRH1 sequence of SEQ ID NO: 55;

(e) a CDRH2 sequence of SEQ ID NO: 56; and

(f) a CDRH3 sequence of SEQ ID NO: 57.

An antibody of the invention may comprise a heavy chain variable region sequence of SEQ ID NO: 60 (the VH of 11878). An antibody of the invention may comprise a light chain variable region sequence of SEQ ID NO: 58 (the VL of 11878). An antibody of the invention may comprise a VH and VL sequence pair of SEQ ID NOs: 60 and 58.

An antibody of the invention may also comprise a VH of SEQ ID NO: 63. This is a humanised sequence of 11878. An antibody of the invention may comprise a VL of SEQ ID NO: 62. This is also a humanised sequence of 11878. In particular an antibody of the invention may comprise a VH of SEQ ID NO: 63 and a VL of SEQ ID NO: 62.

Furthermore, an antibody of the invention may comprise one or more CDR sequences selected from the following:

(a) a CDRL1 sequence of SEQ ID NO: 64;

(b)a CDRL2 sequence of SEQ ID NO: 65;

(c)a CDRL3 sequence of SEQ ID NO: 66;

(d)a CDRH1 sequence of SEQ ID NO: 67, 68 or 69;

(e)a CDRH2 sequence of SEQ ID NO: 70, 71 or 72; and

(f) a CDRH3 sequence of SEQ ID NO: 73.

SEQ ID NOs: 64, 65, 66, 67, 70 and 73 are the parental rabbit antibody CDR sequences of 11883. SEQ ID NOs: 68, 69, 71 and 72 are humanized variants of the rabbit CDRs.

For example, an antibody of the invention may comprise at least one, at least two or all three heavy chain CDR sequences of SEQ ID NOs: 67/68/69, 70/71/72 or 73.

An antibody of the invention may also comprise at least one, at least two or all three light chain CDR sequences of SEQ ID NOS: 64, 65 or 66.

Typically, the antibody of the invention comprises at least one heavy chain CDR sequence selected from SEQ ID NOS: 67-73 and at least one light chain CDR sequence selected from SEQ ID NOS 64-66. The antibody of the invention may comprise at least two heavy chain CDR sequences selected from SEQ ID NOS: 67-73 and at least two light chain CDR sequences selected from SEQ ID NOS: 64-66. In some instances, an antibody of the invention may comprise a HCDR3 of SEQ ID NO: 73.

An antibody of the invention may in particular comprise the following CDRs:

(a) a CDRLl sequence of SEQ ID NO: 64;

(b)a CDRL2 sequence of SEQ ID NO: 65;

(c)a CDRL3 sequence of SEQ ID NO: 66;

(d)a CDRH1 sequence of SEQ ID NO: 67, 68 or 69;

(e)a CDRH2 sequence of SEQ ID NO: 70, 71 or 72; and

(f) a CDRH3 sequence of SEQ ID NO: 73.

For example, an antibody of the invention may comprise the following:

(a) a CDRLl sequence of SEQ ID NO: 64;

(b)a CDRL2 sequence of SEQ ID NO: 65;

(c)a CDRL3 sequence of SEQ ID NO: 66;

(d)a CDRH1 sequence of SEQ ID NO: 67;

(e)a CDRH2 sequence of SEQ ID NO: 70; and

(f) a CDRH3 sequence of SEQ ID NO: 73.

An antibody of the invention may comprise a heavy chain variable region sequence of SEQ ID NO: 76 (the VH of 11883). An antibody of the invention may comprise a light chain variable region sequence of SEQ ID NO: 74 (the VL of 11883). An antibody of the invention may comprise a VH and VL sequence pair of SEQ ID NOs: 76 and 74.

An antibody of the invention may also comprise a VH of SEQ ID NO: 79, 80, 81 or 82. These are humanised sequences of 11883. An antibody of the invention may comprise a VL of SEQ ID NO: 78. This is a humanised sequence of 11883. In particular an antibody of the invention may comprise a VH selected from the group consisting of SEQ ID NOs: 79, 80, 81 and 82 and a VL of SEQ ID NO: 78.

(a) a CDRLl sequence of SEQ ID NO: 83;

(b)a CDRL2 sequence of SEQ ID NO: 84;

(c)a CDRL3 sequence of SEQ ID NO: 85;

(d)a CDRH1 sequence of SEQ ID NO: 86 or 87; (e)a CDRH2 sequence of SEQ ID NO: 88 or 89; and

(f) a CDRH3 sequence of SEQ ID NO: 90.

SEQ ID NOs: 83, 84, 85, 86, 88 and 90 are the parental rabbit antibody CDR sequences of 11889. SEQ ID NOs: 87 and 89 are humanized variants of the rabbit CDRs.

For example, an antibody of the invention may comprise at least one, at least two or all three heavy chain CDR sequences of SEQ ID NOs:86/87, 88/89 or 90.

An antibody of the invention may also comprise at least one, at least two or all three light chain CDR sequences of SEQ ID NOs: 83, 84 or 85.

Typically, the antibody of the invention comprises at least one heavy chain CDR sequence selected from SEQ ID NOs: 86-90 and at least one light chain CDR sequence selected from SEQ ID NOs 83-85. The antibody of the invention may comprise at least two heavy chain CDR sequences selected from SEQ ID NOs: 86-90 and at least two light chain CDR sequences selected from SEQ ID NOS: 83-85. In some instances, an antibody of the invention may comprise a HCDR3 of SEQ ID NO: 90.

An antibody of the invention may in particular comprise the following CDRs:

(a) a CDRLl sequence of SEQ ID NO: 83;

(b)a CDRL2 sequence of SEQ ID NO: 84;

(c)a CDRL3 sequence of SEQ ID NO: 85;

(d)a CDRH1 sequence of SEQ ID NO: 86 or 87;

(e)a CDRH2 sequence of SEQ ID NO: 88 or 89; and

(f) a CDRH3 sequence of SEQ ID NO: 90.

For example, an antibody of the invention may comprise the following:

(a) a CDRLl sequence of SEQ ID NO: 83;

(b)a CDRL2 sequence of SEQ ID NO: 84;

(c)a CDRL3 sequence of SEQ ID NO: 85;

(d)a CDRH1 sequence of SEQ ID NO: 86;

(e)a CDRH2 sequence of SEQ ID NO: 88; and

(f) a CDRH3 sequence of SEQ ID NO: 90.

An antibody of the invention may comprise a heavy chain variable region sequence of SEQ ID NO: 93 (the VH of 11889). An antibody of the invention may comprise a light chain variable region sequence of SEQ ID NO: 91 (the VL of 11889). An antibody of the invention may comprise a VH and VL sequence pair of SEQ ID NOs: 93 and 91. An antibody of the invention may also comprise a VH of SEQ ID NO: 96 or 97.

These are humanised sequences of 11889. An antibody of the invention may comprise a VL of SEQ ID NO: 95. This is a humanised sequence of 11889. In particular an antibody of the invention may comprise a VH selected from the group consisting of SEQ ID NOs:

96 and 97 and a VL of SEQ ID NO: 95.

An antibody of the invention may comprise one or more CDR sequences selected from the following:

(a)a CDRL1 sequence of SEQ ID NO: 98;

(b)a CDRL2 sequence of SEQ ID NO: 99;

(c)a CDRL3 sequence of SEQ ID NO: 100, 101 or 102;

(d)a CDRH1 sequence of SEQ ID NO: 103;

(e)a CDRH2 sequence of SEQ ID NO: 104; and

(f) a CDRH3 sequence of SEQ ID NO: 105, 106 or 107.

SEQ ID NOs: 98, 99, 100, 103, 104 and 105 are the parental rabbit antibody CDR sequences of 11892. SEQ ID NOs: 101, 102, 106 and 107 are humanized variants of the rabbit CDRs.

For example, an antibody of the invention may comprise at least one, at least two or all three heavy chain CDR sequences of SEQ ID NOs: 103, 104 and 105/106/107.

An antibody of the invention may also comprise at least one, at least two or all three light chain CDR sequences of SEQ ID NOs: 98, 99 and 100/101/102.

Typically, the antibody of the invention comprises at least one heavy chain CDR sequence selected from SEQ ID NOs: 103-107 and at least one light chain CDR sequence selected from SEQ ID NOs 98-102. The antibody of the invention may comprise at least two heavy chain CDR sequences selected from SEQ ID NOs: 103-107 and at least two light chain CDR sequences selected from SEQ ID NOs: 98-102. In some instances, an antibody of the invention may comprise a HCDR3 of SEQ ID NO: 105, 106 or 107.

An antibody of the invention may in particular comprise the following CDRs:

(a)a CDRLl sequence of SEQ ID NO: 98;

(b)a CDRL2 sequence of SEQ ID NO: 99;

(c)a CDRL3 sequence of SEQ ID NO: 100, 101 or 102;

(d)a CDRH1 sequence of SEQ ID NO: 103;

(e)a CDRH2 sequence of SEQ ID NO: 104; and (f) a CDRH3 sequence of SEQ ID NO: 105, 106 or 107.

For example, an antibody of the invention may comprise the following:

(a)a CDRL1 sequence of SEQ ID NO: 98;

(b)a CDRL2 sequence of SEQ ID NO: 99;

(c)a CDRL3 sequence of SEQ ID NO: 100;

(d)a CDRH1 sequence of SEQ ID NO: 103;

(e)a CDRH2 sequence of SEQ ID NO: 104; and

(f) a CDRH3 sequence of SEQ ID NO: 105.

An antibody of the invention may comprise a heavy chain variable region sequence of SEQ ID NO: 110 (the VH of 11892). An antibody of the invention may comprise a light chain variable region sequence of SEQ ID NO: 108 (the VL of 11892). An antibody of the invention may comprise a VH and VL sequence pair of SEQ ID NOs: 110 and 108.

An antibody of the invention may also comprise a VH of SEQ ID NO: 115, 116 or 117. These are humanised sequences of 11892. An antibody of the invention may comprise a VL of SEQ ID NO: 112, 113 or 114. These are humanised sequences of 11892. In particular an antibody of the invention may comprise a VH selected from the group consisting of SEQ ID NOs: 115-117 and a VL selected from the group consisting of SEQ ID NO: 112-114.

(a) a CDRL1 sequence of SEQ ID NO: 118;

(b)a CDRL2 sequence of SEQ ID NO: 119;

(c)a CDRL3 sequence of SEQ ID NO: 120, 121 or 122;

(d)a CDRH1 sequence of SEQ ID NO: 123;

(e)a CDRH2 sequence of SEQ ID NO: 124; and

(f) a CDRH3 sequence of SEQ ID NO: 125, 126 and 127.

SEQ ID NOs: 118, 119, 120, 123, 124 and 125 are the parental rabbit antibody CDR sequences of 11881. SEQ ID NOs: 121, 122, 126 and 127 are humanized variants of the rabbit CDRs.

For example, an antibody of the invention may comprise at least one, at least two or all three heavy chain CDR sequences of SEQ ID NOs: 123, 124 and 125/126/127. An antibody of the invention may also comprise at least one, at least two or all three light chain CDR sequences of SEQ ID NOs: 118, 119 and 120/121/122.

Typically, the antibody of the invention comprises at least one heavy chain CDR sequence selected from SEQ ID NOs: 123-127 and at least one light chain CDR sequence selected from SEQ ID NOs 118-122. The antibody of the invention may comprise at least two heavy chain CDR sequences selected from SEQ ID NOs: 123-127 and at least two light chain CDR sequences selected from SEQ ID NOs: 118-122. In some instances, an antibody of the invention may comprise a HCDR3 of SEQ ID NO: 125, 126 or 127.

An antibody of the invention may in particular comprise the following CDRs:

(a) a CDRLl sequence of SEQ ID NO: 118;

(b)a CDRL2 sequence of SEQ ID NO: 119;

(c)a CDRL3 sequence of SEQ ID NO: 120, 121 or 122;

(d)a CDRH1 sequence of SEQ ID NO: 123;

(e)a CDRH2 sequence of SEQ ID NO: 124; and

(f) a CDRH3 sequence of SEQ ID NO: 125, 126 or 127.

For example, an antibody of the invention may comprise the following:

(a) a CDRLl sequence of SEQ ID NO: 118;

(b)a CDRL2 sequence of SEQ ID NO: 119;

(c)a CDRL3 sequence of SEQ ID NO: 120;

(d)a CDRH1 sequence of SEQ ID NO: 123;

(e)a CDRH2 sequence of SEQ ID NO: 124; and

(f) a CDRH3 sequence of SEQ ID NO: 125.

An antibody of the invention may comprise a heavy chain variable region sequence of SEQ ID NO: 130 (the VH of 11881). An antibody of the invention may comprise a light chain variable region sequence of SEQ ID NO: 128 (the VL of 11881). An antibody of the invention may comprise a VH and VL sequence pair of SEQ ID NOs: 130 and 128.

An antibody of the invention may also comprise a VH of SEQ ID NO: 135, 136 or 137. These are humanised sequences of 11881. An antibody of the invention may comprise a VL of SEQ ID NO: 132, 133 or 134. These are humanised sequences of 1188 T In particular an antibody of the invention may comprise a VH selected from the group consisting of SEQ ID NOs: 135, 136 and 137 and a VL selected from the group consisting of SEQ ID NOs: 132, 133 and 134. Also included are antibodies with engineered for example to (i) remove deamidation and glycosylation sites and/or (ii) iso-asp removal and/or (iii) C-terminal lysine removal and/or N-terminal Q to E exchange.

In one example one or more sequences (for example one or more CDRs) provided herein may be modified to remove undesirable residues or sites, such as cysteine residues or aspartic acid (D) isomerisation sites or asparagine (N) deamidation sites.

For example, one or more cysteine residues in any one of the sequences (for example, in any one of the CDRs) may be substituted with another amino acid, such as serine.

In one example, an asparagine deamidation site may be removed from one or more of the sequences (for example, one or more of the CDRs) by mutating the asparagine residue (N) and/or a neighbouring residue to any other suitable amino acid. In one example an asparagine deamidation site such as NG or NS may be mutated, for example to NA orNT.

In one example, an aspartic acid isomerisation site may be removed from one or more of the sequences (for example, one or more of the CDRs) by mutating the aspartic acid residue (D) and/or a neighbouring residue to any other suitable amino acid. In one example an aspartic acid isomerisation site such as DG or DS may be mutated, for example to EG, DA or DT.

In one example, an N-glycosylation site such as NLS may be removed by mutating the asparagine residue (N) to any other suitable amino acid, for example to SLS or QLS.

In one example an N-glycosylation site such as NLS may be removed by mutating the serine residue (S) to any other residue with the exception of threonine (T).

Antibodies of the invention may include a plurality of the above modifications.

The antibody may be or may comprise a variant of one of the specific sequences recited above. For example, a variant may be a substitution, deletion or addition variant of any of the above amino acid sequences.

A variant antibody may comprise 1, 2, 3, 4, 5, up to 10, up to 20 or more (typically up to a maximum of 50) amino acid substitutions and/or deletions from the specific sequences discussed above. “Deletion” variants may comprise the deletion of individual amino acids, deletion of small groups of amino acids such as 2, 3, 4 or 5 amino acids, or deletion of larger amino acid regions, such as the deletion of specific amino acid domains or other features. "Substitution" variants typically involve the replacement of one or more amino acids with the same number of amino acids and making conservative amino acid substitutions. For example, an amino acid may be substituted with an alternative amino acid having similar properties, for example, another basic amino acid, another acidic amino acid, another neutral amino acid, another charged amino acid, another hydrophilic amino acid, another hydrophobic amino acid, another polar amino acid, another aromatic amino acid or another aliphatic amino acid. Some properties of the 20 main amino acids which can be used to select suitable substituents are as follows:

Ala aliphatic, hydrophobic, neutral Met hydrophobic, neutral

Cys polar, hydrophobic, neutral Asn polar, hydrophilic, neutral

Asp polar, hydrophilic, charged (-) Pro hydrophobic, neutral

Glu polar, hydrophilic, charged (-) Gin polar, hydrophilic, neutral

Phe aromatic, hydrophobic, neutral Arg polar, hydrophilic, charged (+)

Gly aliphatic, neutral Ser polar, hydrophilic, neutral

His aromatic, polar, hydrophilic, charged (+) Thr polar, hydrophilic, neutral

He aliphatic, hydrophobic, neutral Val aliphatic, hydrophobic, neutral Lys polar, hydrophilic, charged(+) Trp aromatic, hydrophobic, neutral

Leu aliphatic, hydrophobic, neutral Tyr aromatic, polar, hydrophobic

"Derivatives" or "variants" generally include those in which instead of the naturally occurring amino acid the amino acid which appears in the sequence is a structural analog thereof. Amino acids used in the sequences may also be derivatized or modified, e.g. labelled, providing the function of the antibody is not significantly adversely affected. Derivatives and variants as described above may be prepared during synthesis of the antibody or by post- production modification, or when the antibody is in recombinant form using the known techniques of site- directed mutagenesis, random mutagenesis, or enzymatic cleavage and/or ligation of nucleic acids.

Variant antibodies may have an amino acid sequence which has more than about 60%, or more than about 70%, e.g. 75 or 80%, preferably more than about 85%, e.g. more than about 90 or 95% amino acid identity to the amino acid sequences disclosed herein

(particularly the VH/VL sequences). Furthermore, the antibody may be a variant which has more than about 60%, or more than about 70%, e.g. about 75 or 80%, typically more than about 85%, e.g. more than about 90 or 95% amino acid identity to the VH/VL sequences disclosed herein, whilst retaining the exact CDRs disclosed for these sequences. Variants may retain at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity to the VH/VL sequences disclosed herein (in some circumstances whilst retaining the exact CDRs).

This level of amino acid identity is typically seen across the full length of the relevant SEQ ID NO sequence but may be over a part of the sequence, such as across about 20, 30, 50, 75, 100, 150, 200 or more amino acids, depending on the size of the full length polypeptide.

In connection with amino acid sequences, "sequence identity" refers to sequences which have the stated value when assessed using ClustalW (Thompson et al., 1994, supra) with the following parameters:

Pairwise alignment parameters -Method: accurate, Matrix: PAM, Gap open penalty: 10.00, Gap extension penalty: 0.10;

Multiple alignment parameters -Matrix: PAM, Gap open penalty: 10.00, % identity for delay: 30, Penalize end gaps: on, Gap separation distance: 0, Negative matrix: no, Gap extension penalty: 0.20, Residue-specific gap penalties: on, Hydrophilic gap penalties: on, Hydrophilic residues: GPSNDQEKR. Sequence identity at a particular residue is intended to include identical residues which have simply been derivatized.

The present invention thus provides antibodies having specific sequences and variants which maintain the function or activity of the antibody.

With regards to function, in some instances antibodies of the invention are able to neutralise at least one biological activity of Ebola virus (a neutralising antibody), particularly to neutralise virus infectivity. The ability of an antibody to neutralise virus infectivity may be measured using an appropriate assay, particularly using a cell-based neutralisation assay. In the invention, neutralisation may be determined using an assay for measuring infection of cells using a surrogate virus coated with the Ebola virus glycoprotein. For these surrogate viruses infection of cells is dependent on the Ebola virus glycoprotein.

An example of such an assay uses the E-S-FLU Ebola virus surrogate as described in the Examples below. This assay utilises a disabled influenza virus core coated with Ebola virus GP. The E-S-FLU encodes eGFP that replaces the hemagglutinin coding sequence so that infected cells fluoresce green. The loss of fluorescent signal e.g. after overnight infection provides a measure of the inhibition of infection by an antibody.

In a neutralisation assay, antibodies of the invention may be “partial” neutralising antibodies, where inhibition of infection plateaus at 50-90% inhibition or “strong” neutralising antibodies, which achieve > 90% inhibition. Antibody concentrations may be as tested in the Examples/Figures, for example with an concentration of 5 pg/ml used to determine if any antibody is a “strong” or “partial” neutralising antibody. Neutralisation may also be determined up to a maximum concentration of 50 pg/ml.

As shown in Figures 2 and 3, the 11886 antibody is a strong neutralising antibody for all of Zaire, Sudan and Bundibugyo Ebola virus.

Antibodies of the invention may have sequences as described above and be either a strong or partial neutralising antibody. For example, an antibody may have six CDR sequences of 11886 and be a strong neutralising antibody (preferably for all of Zaire,

Sudan and Bundibugyo Ebola virus).

Neutralisation may also be determined using IC50 or IC90 values. IC50 and IC90 values can be determined from the results of a neutralisation assay (as discussed above) using standard methods. An antibody of the invention may for example have an IC50 value of less than (i.e. better than) 10 pg/ml, less than 5 pg/ml, less than 2 pg/ml or less than 1 pg/ml (typically down to 0.1 pg/ml). In some instances an antibody of the invention may have an IC50 value of between 0.1 pg/ml and 10 pg/ml, sometimes between 0.1 pg/ml and 5 pg/ml, between 0.1 pg/ml and 2 pg/ml or even between 0.1 pg/ml and 1 pg/ml. In some instances, an antibody of the invention may have an IC50 value of between 0.5 pg/ml and 10 pg/ml, sometimes between 0.5 pg/ml and 5 pg/ml or between 0.5 pg/ml and 2 pg/ml.

An antibody of the invention may have an IC90 value of less than 10 pg/ml, optionally less than 5 pg/ml (typically down to 1 pg/ml). For example, an antibody of the invention may have an IC90 value of between 1 pg/ml and 10 pg/ml, for example between 1 pg/ml and 5 pg/ml.

Neutralisation ability may be determined for any species of Ebola virus, such as Zaire, as shown in the Examples. Typically, an antibody of the invention will have the above IC50/90 values for all of Zaire, Sudan and Budibugyo. An antibody of the invention may have an IC50 value of less than 2 pg/ml for Zaire, less than 2 pg/ml (or preferably less than 1 pg/ml) for Sudan and/or less than 1 pg/ml for Budibugyo. In some instances, an antibody of the invention may therefore have an IC50 value of less than 2 pg/ml for all three species. Typical lower limits are as described above.

An antibody of the invention may also have an IC90 of less than 10 pg/ml for Zaire, less than 5 pg/ml for Sudan and less than 5 (or 3 or 2) pg/ml for Budibugyo. An antibody of the invention may therefpre have an IC90 value of less than 10 pg/ml for all three species. Once again, typical lower limits are as described above.

These IC₅₀/ IC₉₀ values may be applied to the sequences described above. For example, an antibody of the invention may have six CDR sequences as described above (particularly the CDRs of 11886) and an IC₅₀/ IC₉₀ value as presented above.

Antibodies of the invention are also preferably able to provide in vivo protection in Ebola virus infected animals. For example, administration of an antibody of the invention to Ebola virus infected animals may result in a survival rate of greater than 30% or greater than 50%. Ideally, antibodies of the invention achieve a survival rate of 100%. Survival rates may be determined using routine methods. For example, in in vivo protection may be determined in mice (such as after a dose of 100 pg antibody at day two of infection). In vivo protection may also be determined in guinea pigs (for example, at a dose of 10 mg/kg of each antibody at day three of infection).

As discussed above, antibodies of the invention are typically cross-reactive for one or more Ebola virus species, such as Zaire (e.g. Zaire Mayinga and/or Makona), Bundibugyo and Sudan (e.g. Sudan Gulu). Antibodies of the invention are also typically cross-reactive for Tai Forest. Preferably, antibodies are cross-reactive for all of the above. In other words, the antibodies are capable of binding to the glycoprotein from these species/strains. Binding can be measured, for example, using Surface Plasmon Resonance. An antibody may be cross-reactive if it retains 100% of its binding capability. An antibody may also be cross-reactive with lower retention of binding, such as retaining at least 50% or at least 30% binding capability across one or all species.

A measure of binding would be the K_D value. For example, antibodies of the invention may be cross-reactive if they have a K_D value of less than 1 pM for more than one species (antibodies may have a K_D value of less than 1 mM for more than one species, such as Zaire, Bundibugyo, Tai Forest and Sudan).

Antibodies of the invention may have any combination of one or more of the above properties.

Antibodies of the invention may bind to the same epitope, or compete for binding to Ebola virus glycoprotein, with any one of the reference antibodies described above (i.e. in particular with antibodies with the heavy and light chain variable regions described above). Methods for identifying antibodies binding to the same epitope, or cross-competing with one another, are discussed below.

Nucleic acids, vectors, host cells and methods of producing antibodies

The present invention also provides an isolated nucleic sequence encoding the heavy and/or light chain variable regions(s) of an antibody molecule of the present invention, or the full heavy and/or light chain (in some instances a pair of nucleic acids encoding the heavy and light chain variable regions or full heavy/light chains).

Nucleic acid sequences which encode an antibody molecule of the present invention may be DNA or RNA (for example mRNA).

Nucleic acid sequences which encode an antibody molecule of the present invention can be obtained by methods well known to those skilled in the art. For example, nucleic acids sequences coding for part or all of the antibody heavy and light chains may be synthesised as desired from the corresponding amino acid sequences.

The invention also provides expression vectors comprising the nucleic acid sequences(s).

Also provided is a host cell comprising one or more cloning or expression vectors comprising one or more nucleic acid sequences encoding an antibody of the present invention. Any suitable host cell/vector system may be used for expression of the nucleic acid sequences encoding the antibody molecule of the present invention. Bacterial, for example E. coli, and other microbial systems may be used or eukaryotic, for example mammalian, host cell expression systems may also be used. Suitable mammalian host cells include CHO, or myeloma. Typically, antibodies may be produced in CHO cells, modified CHO cells (to produce afucosylated antibodies) or HEK-293 cells. The present invention also provides a process for the production of an antibody molecule according to the present invention comprising culturing a host cell containing a vector of the present invention under conditions suitable for leading to expression of protein from DNA encoding the antibody molecule of the present invention, and isolating the antibody molecule.

General methods by which the vectors may be constructed, transfection methods and culture methods are well known to those skilled in the art. In this respect, reference is made to “Current Protocols in Molecular Biology”, 1999, F. M. Ausubel (ed), Wiley Interscience, New York and the Maniatis Manual produced by Cold Spring Harbor Publishing.

Pharmaceutical composition

The invention also provides a pharmaceutical composition comprising an antibody of the invention and a pharmaceutically acceptable carrier or diluent. In some instances, a pharmaceutical composition of the invention may comprise one or more nucleic acids (as described above) encoding an antibody of the invention. The nucleic acids may for example be mRNA.

The pharmaceutical composition may comprises one or more additional anti-Ebola virus antibodies, or sequences encoding one or more additional anti-Ebola virus antibodies. It is preferable that the antibodies do not cross-compete with one another, particularly that the antibodies bind to non-overlapping epitopes on the Ebola virus glycoprotein.

Numerous methods may be used to determine whether antibodies cross-compete or bind to non-overlapping epitopes. Such methods are utilised in the Examples and are discussed further below.

A pharmaceutical composition of the invention may comprise any of the antibodies described above, alone or in any combination (or sequences encoding any of the antibodies described above).

As used herein, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like that are physiologically compatible.

The pharmaceutical compositions of the invention may include one or more pharmaceutically acceptable salts. A "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the parent compound and does not impart any undesired toxicological effects. Examples of such salts include acid addition salts and base addition salts.

Pharmaceutically acceptable carriers comprise aqueous carriers or diluents. Examples of suitable aqueous carriers that may be employed in the pharmaceutical compositions of the invention include water, buffered water and saline. Examples of other carriers include ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. In many cases, it will be desirable to include isotonic agents, for example, sugars, polyalcohols such as mannitol, sorbitol, or sodium chloride in the composition.

Therapeutic compositions typically must be sterile and stable under the conditions of manufacture and storage. The composition can be formulated as a solution, microemulsion, liposome, or other ordered structure suitable to high drug concentration.

Pharmaceutical compositions of the invention may comprise additional therapeutic ingredients, for example additional anti-viral agents. Anti-viral agents may bind to Ebola virus and inhibit viral activity. Alternatively, anti-viral agents may not bind directly to Ebola virus but still affect viral activity/infectivity. An anti-viral agent could be a further anti-Ebola antibody, which binds somewhere other than the glycoprotein. The additional therapeutic ingredient could also be an anti-inflammatory agent, such as a corticosteroid or a non-steroidal anti-inflammatory drug. The additional therapeutic agent could also be an anti-Ebola vaccine.

The pharmaceutical composition may be administered subcutaneously, intravenously, intradermally, intramuscularly, intranasally or orally.

Antibody cocktails

The invention also provides anti-Ebola virus antibody cocktails, particularly a cocktail comprising two or more, typically three or more antibodies to the Ebola virus glycoprotein. As used herein, an “antibody cocktail” generally refers to a combination/mixture of antibodies within the same composition, i.e. a single pharmaceutical composition comprising the antibodies. However, the invention also includes the combined use of different anti-Ebola virus antibodies in separate pharmaceutical compositions. Once again the pharmaceutical compositions may comprise the antibodies per se, or nucleic acid sequences (e.g. mRNA) encoding the antibodies.

The antibodies may bind to the Ebola virus glycoprotein from any of the species/strains discussed above.

A cocktail of the invention may comprise two or more antibodies (or sequences encoding the antibodies). Typically, a cocktail of the invention comprises two or more antibodies binding to different regions of the Ebola virus glycoprotein. For example, a cocktail may comprise two or more antibodies binding to at least two of the following regions of the glycoprotein: glycan cap, b17-18 loop, receptor binding region and base. In some instances, a cocktail of the invention may comprise three or four antibodies binding to the Ebola virus glycoprotein.

The skilled person would readily be able to determine the amino acid numbering for each of these domains of the glycoprotein based on the published information for the various species/strains.

The skilled person would also readily be able to determine the binding site (epitope) of an antibody using standard techniques, such as those described above. The skilled person could also readily determine whether an antibody binds to the same epitope as, or competes for binding with, a reference antibody by using routine methods known in the art.

For example, to determine if a test antibody binds to the same epitope as a reference antibody of the invention, the reference antibody is allowed to bind to a protein or peptide under saturating conditions. Next, the ability of a test antibody to bind to the protein or peptide is assessed. If the test antibody is able to bind to the protein or peptide following saturation binding with the reference antibody, it can be concluded that the test antibody binds to a different epitope than the reference antibody. On the other hand, if the test antibody is not able to bind to protein or peptide following saturation binding with the reference antibody, then the test antibody may bind to the same epitope as the epitope bound by the reference antibody of the invention.

To determine if an antibody competes for binding with a reference antibody, the above-described binding methodology is performed in two orientations. In a first orientation, the reference antibody is allowed to bind to a protein/peptide under saturating conditions followed by assessment of binding of the test antibody to the protein/peptide molecule. In a second orientation, the test antibody is allowed to bind to the protein/peptide under saturating conditions followed by assessment of binding of the reference antibody to the protein/peptide. If, in both orientations, only the first (saturating) antibody is capable of binding to the protein/peptide, then it is concluded that the test antibody and the reference antibody compete for binding to the protein/peptide. As will be appreciated by the skilled person, an antibody that competes for binding with a reference antibody may not necessarily bind to the identical epitope as the reference antibody, but may sterically block binding of the reference antibody by binding an overlapping or adjacent epitope.

Two antibodies bind to the same or overlapping epitope if each competitively inhibits (blocks) binding of the other to the antigen. That is, a 1-, 5-, 10-, 20- or 100-fold excess of one antibody inhibits binding of the other by at least 50%, 75%, 90% or even 99% as measured in a competitive binding assay (see, e.g., Junghans et ak, Cancer Res,

1990:50: 1495-1502). Alternatively, two antibodies have the same epitope if essentially all amino acid mutations in the antigen that reduce or eliminate binding of one antibody reduce or eliminate binding of the other. Two antibodies have overlapping epitopes if some amino acid mutations that reduce or eliminate binding of one antibody reduce or eliminate binding of the other.

Additional routine experimentation (e.g., peptide mutation and binding analyses) can then be carried out to confirm whether the observed lack of binding of the test antibody is in fact due to binding to the same epitope as the reference antibody or if steric blocking (or another phenomenon) is responsible for the lack of observed binding. Experiments of this sort can be performed using ELISA, RIA, surface plasmon resonance, flow cytometry or any other quantitative or qualitative antibody-binding assay available in the art.

Other techniques that may be used to determine antibody epitopes include hydrogen/deuterium exchange, X-ray crystallography and peptide display libraries (as described in the Examples). A combination of these techniques may be used to determine the epitope of the test antibody.

In the antibody cocktails of the invention it is preferred that the antibodies bind non-overlapping epitopes, or do not cross-compete with one another. This can be determined using the methods described above. One or more of the antibodies included in the cocktails of the invention may be neutralising antibodies (in other words, one or more of the antibodies may be individually neutralising). In some instances, all of the antibodies in the cocktail may be neutralising antibodies. Such neutralising antibodies are described above.

It is also typical that one or more of the antibodies in the cocktail individually enhance survival of animals infected with Ebola virus. In some instances, all of the antibodies in the cocktail enhance survival of Ebola virus infected animals.

Typically, administration of an antibody cocktail of the invention to Ebola virus infected animals results in a survival rate of at least 50%. Preferably, administration of an antibody cocktail of the invention to Ebola virus infected animals results in a 100% survival rate. For example, survival rates may be determined in mice (e.g. with a single dose of 100 pg of antibody at day 2 of infection). Survival rates may also be determined in infected guinea pigs. Antibodies may be administered at day three following infection. Such experiments may be conducted at a dose of 10 mg/kg of each antibody, or in some instances at a total dose (for all antibodies) of 5 mg/kg. A cocktail may therefore result in a survival rate of at least 50% at a dose of 10 mg/kg of each antibody or at a total dose of 5 mg/kg. A cocktail may result in a 100% survival rate at a dose of 10 mg/kg of each antibody or at a total dose of 5 mg/kg.

Furthermore, it is advantageous if one or more (for example, one, two, three or four) antibodies in the cocktail cross-react with different Ebola virus species, for example Zaire and/or Sudan and/or Bundibugyo. It is most preferred that all of the antibodies in the cocktail cross-react with all of these species. Cross-reactivity is discussed above.

The cocktail may include any of the antibodies of the invention as defined above.

For example, the cocktail of the invention may include an antibody based on 11886 (i.e. with the sequences described above). In this scenario, additional antibodies included in the cocktail could be one or more antibodies selected from 66-3-9C, 040 and 6662.

For example an antibody in the cocktail may comprise one or more of the following CDR sequences:

(a) a CDRL1 sequence of SEQ ID NO: 141, 168, 169 or 170;

(b)a CDRL2 sequence of SEQ ID NO: 142;

(c)a CDRL3 sequence of SEQ ID NO: 143;

(d)a CDRH1 sequence of SEQ ID NO: 138; (e)a CDRH2 sequence of SEQ ID NO: 139; and

(f) a CDRH3 sequence of SEQ ID NO: 140.

These are CDR sequences of the 66-3-9C antibody. Typically the antibody comprises six CDRs as set out above.

An antibody in the cocktail may also comprise a VH of SEQ ID NO: 144 and/or a VL of SEQ ID NO: 145. These are the VH and VL sequences of the 66-3 -9C antibody. Furthermore, an antibody may comprise a heavy chain of SEQ ID NO: 162 and a light chain of SEQ ID NO: 163. These are heavy and light chain sequences of 66-9-3 C.

An antibody in the cocktail may also comprise one or more of the following CDR sequences:

(a) a CDRLl sequence of SEQ ID NO: 149;

(b)a CDRL2 sequence of SEQ ID NO: 150;

(c)a CDRL3 sequence of SEQ ID NO: 151;

(d)a CDRH1 sequence of SEQ ID NO: 146;

(e)a CDRH2 sequence of SEQ ID NO: 147; and

(f) a CDRH3 sequence of SEQ ID NO: 148.

These are CDR sequences of the 040 antibody. Typically the antibody comprises six CDRs as set out above.

An antibody in the cocktail may also comprise a VH of SEQ ID NO: 152 and/or a VL of SEQ ID NO: 153. These are the VH and VL sequences of the 040 antibody. Furthermore, an antibody may comprise a heavy chain of SEQ ID NO: 164 and a light chain of SEQ ID NO: 165. These are heavy and light chain sequences of 040.

(a) a CDRLl sequence of SEQ ID NO: 157;

(b)a CDRL2 sequence of SEQ ID NO: 158;

(c)a CDRL3 sequence of SEQ ID NO: 159, 176, 177 or 178;

(d)a CDRH1 sequence of SEQ ID NO: 154;

(e) a CDRH2 sequence of SEQ ID NO: 155, 171, 172 or 173; and

(f) a CDRH3 sequence of SEQ ID NO: 156, 174 or 175.

These are CDR sequences of the 6662 antibody. Typically the antibody comprises six CDRs as set out above. An antibody in the cocktail may also comprise a VH of SEQ ID NO: 160 and/or a VL of SEQ ID NO: 161. These are the VH and VL sequences of the 6662 antibody. Furthermore, an antibody may comprise a heavy chain of SEQ ID NO: 166 and a light chain of SEQ ID NO: 167. These are heavy and light chain sequences of 6662.

These sequences may be modified as discussed above.

In some instances, an antibody of the invention may be used in a cocktail in combination with an antibody based on 66-3-9C, 040 and 6662 (i.e. with the above sequences).

The antibodies may be formulated using a pharmaceutically acceptable carrier or diluent, as discussed above.

Therapeutic uses

The antibodies, nucleic acids, pharmaceutical composition and cocktails of the invention may be used for the treatment, prevention or amelioration of Ebola virus infection. In other words, the antibodies may be used for the treatment of disease associated with Ebola virus and/or to decrease the viral load. Ebola virus disease develops after infection with Ebola virus and the subsequent incubation period. Early symptoms of Ebola virus infection are fatigue fever, myalgia, headache, sore throat, which are followed by vomiting, diarrhoea, exanthema, renal and hepatic dysfunction, external haemorrhage and other symptoms. Antibodies, pharmaceutical composition and cocktails of the invention may be used to ameliorate or reduce the severity, duration or frequency of one or more symptoms associated with Ebola virus infection. The symptom may be fever, headache, fatigue, loss of appetite, myalgia, diarrhoea, vomiting, abdominal pain, dehydration and/or bleeding.

As described above, the invention comprises not only the administration of antibodies themselves, but also nucleic acid sequences encoding the antibodies (typically mRNA).

Typically, the invention relates to the administration of the antibodies/compositions to a human subject in need thereof. However, administration to non-human animals such as rats, rabbits, sheep, pigs, cows, cats, dogs is also contemplated. The subject may be at risk of exposure to Ebola virus infection, such as a healthcare worker or a person who has come into contact with an infected individual. A subject may have visited or be planning to visit a country known or suspected of having an Ebola outbreak. A subject may also be at greater risk, such as an immunocompromised individual (for example an individual receiving immunosuppressive therapy or an individual suffering from human immunodeficiency syndrome (HIV) or acquired immune deficiency syndrome (AIDS).

The antibodies, nucleic acids, compositions and cocktails of the invention may be administered therapeutically or prophylactically.

As discussed above, the antibodies, nucleic acids, pharmaceutical compositions and cocktails may be administered subcutatneously, intravenously, intradermally, orally, intranasally, intramuscularly or intracranially, Typically, the antibodies, nucleic acids, pharmaceutical compositions and cocktails are administered intravenously or subcutaneously.

The dose of an antibody may vary depending on the age and size of a subject, as well as on the disease, conditions and route of administration. Antibodies may be administered at a dose of about 0.1 mg/kg body weight to a dose of about 100 mg/kg body weight, such as at a dose of about 5 mg/kg to about 10 mg/kg. Antibodies may also be administered at a dose of about 50 mg/kg, 10 mg/kg or about 5 mg/kg body weight.

A cocktail of the invention may for example be administered at a dose of about 5 mg/kg to about 10 mg/kg for each antibody, or at a dose of about 10 mg/kg or about 5 mg/kg for each antibody. Alternatively, a cocktail may be administered at a dose of about 5 mg/kg total (e.g. a dose of 1.67 mg/kg of each antibody in a three antibody cocktail).

The initial dose may be followed by administration of a second or plurality of subsequent doses. The second and subsequent doses may be separated by an appropriate time.

As discussed above, the antibodies of the invention (or sequences encoding antibodies of the invention) are typically used in a single pharmaceutical composition/cocktail (co-formulated). However, the invention also generally includes the combined use of antibodies of the invention (in separate preparations/compositions). “In combination with” means that a first antibody may be administered prior to, concurrent with or after a second (or subsequent) antibody. “Concurrent” with includes administration both in single and separate dosage forms, where such separate dosage forms may be administered e.g. within 30 minutes or less of one another. “Prior to” may include administration e.g. one week before, 48 hours before or 24 hours before. “After” may include e.g. 24 hours after, 48 hours after, or 72 hours after. The dosage forms may be administered by the same route, or by different routes. “In combination with” also includes sequential or concomitant administration.

The same applies for administration of additional therapeutic agents, which are discussed above. These may be administered in combination with antibodies/nucleic acids/pharmaceutical compositions/cocktails of the invention.

The following examples are presented below so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the invention. The examples are not intended to limit the scope of what the inventors regard as their invention.

Examples

Example 1: Antibody discovery rabbit immunisations

Adherent rabbit fibroblast cell line cultured in 5-stack CellSTACK® Culture Chambers (Corning, 3319) were lifted using StemPro™ Accutase™ Cell Dissociation Reagent (Gibco, A1110501), washed and resuspended in Earle’s Balanced Salts (Sigma, E3024). Cells were transfected using electroporation (160-170V, 20 seconds, 5 amps) with 3x10^L7 cells in 600 pL per GenePulser electroporation cuvette (BioRad, #1652088). Cells were transfected with EBOV GP (NP 066246.1), SUDV GP (YP 138523.1), BDBV GP (YP_003815435.1) or TAFV GP (YP_003815426.1). Cells were recovered in 50 mL media (RPME 10% FBS /2mM glutamine), transferred to T175 tissue culture flask and incubated at 37°C, 5% CO2 overnight. Cells were lifted the next day, counted and cell viability measured. Expression of antigens was confirmed by FACS via staining with human antigen-specific antibody and goat anti-human Fc-specific AffmiPure F(ab')₂ Fragment AlexaFluor-647 conjugate (Jackson 109-606-170). Fluorescence was measured using BD FACS CantoTM 11 and analysed with FlowJo3 software. Cells were frozen down prior to formulation in for vaccination and stored in LN2.

One female New Zealand White rabbit was immunised four times subcutaneously at two week intervals with rabbit fibroblast cells expressing Ebola virus glycoproteins. Each dose consisted of 2.5xl0⁶ cells transfected with EBOV GP, SUDV GP, BDBV GP and TAFV GP (lxlO⁷ cells/dose total) in 500 mE PBS. First dose only was adjuvanted with complete Freund’s adjuvant delivered at a separate site. Serum was taken on day of each vaccination and presence of antigen specific IgG confirmed by binding of serum IgG to GPs transiently transfected HEK cells assessed using an iQUE flow cytometer system (Intellicyt).

Example 2: isolation of mAbs from memory B cells

Lymph nodes were harvested 14 days after final vaccination. B cell culture screening was performed using a method similar to that described by Tickle et al. (Tickle et al. 2015, J Biomol Screen, 20: 492-7). Rabbit B cell cultures were prepared using 50 x 96-well plates at a cell density of approximately 5000 cells per well. After 6 days culture, screening was performed.

Briefly, the presence of Ebola virus glycoprotein-binding antibodies in B cell culture supernatants was determined using a homogeneous fluorescence-based binding assay performed on a Mirrorball® fluorescence cytometer device using MDCK cells stably transfected to express surface glycoprotein from EBOV or SUDV B cell supernatants were simultaneously counter screened for binding to the parental MDCK SIAT-1 cells. EBOV and SUDV GP expressing cells were stained with DiO and untransfected parental cells were stained with Dil (Vybrant Multicolour cell-labelling kit, Invitrogen, V22889). Binding of B cell culture supernatant was revealed with Alexa Fluor® 647 AffmiPure F(ab')₂ Fragment Rabbit Anti-Human IgG, Fey fragment specific antibody (Jackson, 309- 606-008).

Following primary screening, positive supernatants containing GP -reactive antibody were consolidated on 96-well bar-coded master plates and B cells in cell culture plates frozen at -80°C. Master plates were then screened in a further homogeneous fluorescence binding assay to confirm that the antibodies bound the Ebola virus glycoprotein-expressing MDCK-SIATl cells and not the parental MDCK-SIAT1 cells, as well as to identify antibodies that bound to both EBOV and SUDV GPs. Briefly, a homogeneous fluorescence-based binding assay was performed on an Applied Biosystems 8200 cellular detection system device using parental MDCK SIAT-1 cells, and stably transfected EBOV GP and SUDV GP expressing MDCK SIAT-1 cell lines thawed from aliquots stored in LN2. Binding was revealed with a Alexa Fluor® 647 AffmiPure F(ab')₂ Fragment Goat Anti-Rabbit IgG, Fc fragment specific antibody (Jackson, 111-606-046). Additionally, supernatants were screened in a flow cytometric assay using suspension ExpiHEK293 cells transiently transfected to express EBOV GP or SUDV GP. This was to confirm that binding to the GP expressed on the MDCK SIAT-1 cell lines had not been due to changes to the GP caused by trypsin or Accutase digestion used to lift the adherent cells from tissue culture flasks for use in the previous binding assays.

Briefly, ExpiHEK293 cells were transfected two days prior to use in the binding assay using ExpiFectamine 293 (Invitrogen) with EBOV GP, SUDV GP or an irrelevant antigen podoplanin as a negative control. Transfection enhancers were added the next day. On the third day, expression of surface Ebola virus glycoproteins was confirmed using known broadly-reactive Ebola virus GP human mAh and Alexa Fluor® 647 AffmiPure F(ab')₂ Fragment Goat Anti-Human IgG, Fey fragment specific antibody (Jackson, 109- 606-170). Expression of podoplanin was confirmed with an Anti -Podoplanin mAh PE conjugate (Biolegend, 337004). After confirmation of antigen expression, approximately 10,000 transiently transfected cells/well were incubated with 10 pL B cell culture supematant/well in 384 well plates. After washing, cells were incubated with Alexa Fluor® 647 AffmiPure F(ab')₂ Fragment Goat Anti-Rabbit IgG, Fc fragment specific antibody (Jackson, 111-606-046). After washing, fluorescence was detected using an iQUE flow cytometer system (Intellicyt).

The Fluorescent Foci method (US Patent 7993864/ Europe EP1570267B1; (Clargo et al. 2014, MAbs, 6: 143-59) utilizing soluble recombinant EBOV GP with deleted transmembrane domain (GPATM) conjugated directly to 2-2.9 pm diameter SPHEROTM polystyrene magnetic beads (Spherotech, PP -20-100) and soluble recombinant biotinylated SUDV GPATM conjugated to 0.1 pm diameter SuperAvidin™ Coated Microspheres (Bangs Laboratories, Inc. CPOIN). SUDV GPATM was biotinylated using EZ-Link Sulfo-NHS-Biotin kit (Thermo Scientific, 21326).

This method was used to identify and isolate antigen-specific B cells from positive wells with the different diameter beads allowing for identification of wells with foci formed as a result of binding to both antigens. Specific antibody variable region genes were recovered from single cells by reverse transcription (RT)-PCR using heavy and light chain variable region-specific primers. PCR primers contained restriction sites at the 3’ and 5’ ends allowing cloning of the variable region into a human IgGl (VH), human kappa (VK) or human lambda (Vk) mammalian expression vector. Heavy and light chain constructs were co-transfected into Expi293F cells using Expifectamine 293 (Invitrogen) and recombinant antibody expressed. After 5 days expression, supernatants were harvested and antibody rescreened for selectivity using the specificity assays described above.

Example 3: Purification of antibodies

Antibody was purified from cell culture supernatant via Protein A affinity chromatography utilising an AKTA Pure system (GE Healthcare) with CETAC autosampler. Antibodies were purified from 30 mL of culture supernatant using 2ml HiTrap MabSelect SuRe Protein A column (GE Healthcare, Life Sciences) and eluted in 0.1M sodium citrate, pH 3.4. Eluted fractions were neutralised with 2 M Tris/HCl pH 8.0. Pooled peak fractions were buffer exchanged into PBS pH 7.4 and concentrated using Amicon Ultra Spin columns with a 30KDa cut off membrane (Millipore, UFC905008) and centrifugation at 4000 g, before sterile filtering. Concentration was measured by A280 (Nanodrop spectrophotometer). Monomer purity was assessed by size exclusion on a UPLC (Acquity) with a BEH200; 1.7 mM, 4.6 mm X 300 mm column (Waters,

176003905) and developed with an isocratic gradient of 0.2 M phosphate, pH 7.0 at 0.3 mL/min.

Example 4: Binding of antibodies to Ebola virus glycoproteins

As previously, ExpiHEK293 cells were transfected two days prior to use in the binding assay using ExpiFectamine 293 (Invitrogen) with EBOV GP, SUDV GP, BDBV GP or TAFV GP (same strains as used for immunisations for antibody generation) or an irrelevant antigen podoplanin as a negative control. Transfection enhancers were added the next day. On the third day, expression of surface Ebola virus glycoproteins was confirmed using known broadly-reactive Ebola virus GP human mAh and Alexa Fluor® 647 AffmiPure F(ab')₂ Fragment Rabbit Anti -Human IgG, Fey fragment specific antibody (Jackson, 309-606-008). Expression of podoplanin was confirmed with an Anti- Podoplanin mAh PE conjugate (Biolegend, 337004). Assay was conducted in 384-well plates with approximately 5,000 transiently transfected cells/well. Cells were incubated with 30 pL of titrated purified rabbit monoclonal antibody starting at 10 pg/mL. After washing, cells were incubated with Alexa Fluor® 647 AffmiPure F(ab')₂ Fragment Goat Anti-Rabbit IgG, Fc fragment specific antibody (Jackson, 111-606-046). After washing, fluorescence was detected using an iQUE flow cytometer system (Intellicyt).

Results are presented in Figure 1. All broadly reactive mAbs showed titratable binding to all four species of GP tested and no reactivity to mock cells transfected with an irrelevant antigen.

Example 5: In vitro Ebola virus glycoprotein pseudotyped S-FLU virus microneutralisation assay

This assay measures the ability of an antibody to prevent the infection of MDCK SIAT-1 cells by an S-FLU pseudotype virus coated with GP from an Ebola virus (Xiao, J., et al, J Virol., 2018 Jan 30;92(4)). The virus is replication incompetent and contains no viral RNA or DNA encoding the GP allowing it to be handled at BSL-2. A GFP reporter gene in the virus replaces the hemagglutinin coding sequence, making infected MDCK SIAT-1 cells detectable by fluorescence. Loss of fluorescence signal indicates neutralisation of the virus and prevention of infection.

In a 96 well, flat bottom tissue culture plate, virus diluted in 50 pL viral growth media (VGM: DMEM, 0.1% BSA, lOmM HEPES, Penicillin/Streptomycin 100U each, 2mM Glutamine) was incubated with antibody diluted 50 pL in phosphate buffered saline (PBS) at titrated concentration for 2 hours, 37°C, 5% C02, before addition of 100 pL MDCK SIAT-1 cells in VGM (3c10^L5 cells/mL). Virus was used at a dilution previously determined to give maximum infection of cells. After 20-24 hours incubation at 37°C, 5% CO2, supernatant was aspirated from cells, and cells fixed with 100 pL 10% formalin for 30 minutes, 4°C. Formalin was removed and 100 pL PBS added to each well. GFP fluorescence was read using a Clariostar plate reader (BMG Labtech). Maximum infection was indicated by signal from cells infected with viruses pre-incubated with PBS only or a non-GP binding antibody. Minimum signal was indicated by uninfected cells. Inhibitory concentration at 50% (IC50) and 90% (IC90) was derived by linear interpolation.

Results are presented in Figures 2 and 3. Example 6: Peptide phage display

The panel of novel broadly reactive monoclonal antibodies were used to pan two different random combinatorial linear peptide libraries. Peptide sequences enriched by each mAb were analysed and motifs aligned with GP sequences to identify potential partial epitopes. In each experiment two rounds of panning against each mAb was conducted with decreasing concentrations of antibody. Round 2 panning also included a subtractive step where phage were incubated with non-GP specific rabbit IgG and unbound phage transferred to incubate with the broadly reactive GP mAb. All panning steps occurred with mAb adhered to a plastic surface. After Round 2, phagemid DNA sequences encoding peptide sequences were barcoded via PCR and subjected to Next Generation Sequencing (NGS).

9mer and 13mer random combinatorial linear p8 phage peptide libraries were available.

Biopanning was carried out with both libraries against each monoclonal antibody with two rounds of selection and additional subtractive panning against species relevant IgG to reduce amplification of phage bound to portions of the antibodies other than the CDRs. A control human antibody with known GP epitope previously identified via yeast display technologies was used to pan phage libraries in parallel to rabbit monoclonal antibodies to validate method.

In brief, 3-8 wells of 96 well NUNC MAXISORP plates were coated with 1 5pg of monoclonal antibody in PBS and incubated overnight at 4°C. Plates were washed twice with 0.1%Tween/PBS. Plates were blocked with 3% milk powder/PBS at room temperature for at least one hour. Phage library aliquot was thawed and incubated in approximately 2.5 times volume of 6% milk powder/PBS, at room temperature with mixing for at least one hour. Plates were washed twice with 0. l%Tween/PBS. Each mAb coated well was incubated with 100pL of blocked phage (~2.5c10^L13 cfu/mL), at room temperature for one hour with gentle shaking. Plates were washed ten times with 0.1%T ween/PBS. Bound phage were eluted in 0.1 M HC1 (Sigma, 2104) and neutralised with 1 M Tris-HCl pH 8.0 (Sigma, T2694). Eluted phage (Rl) were used to infect TGI E.coli (F¹ traD36 proAB lacIqZ DM15] supE thi-1 A(lac-proAB) A(mcrB-hsdSM)5(rK - mK -))(Lucigen, 60502) cells and grown on 2TY plates supplemented with carbenicillin and glucose at 30°C. R1 output bacteria were scraped (a small aliquot of each was flash frozen and stored at -80°C), grown at 37°C to OD-0.5 and rescued using M13K07 helper phage (NEB, N0315S). Rescued cultures were resuspended in 2TY- Carbenicillin/Kanamycin without glucose and grown overnight at 30°C to produce phage for R2 panning. Next day phage were precipitated using 20% PEG80002.5 M NaCl, resuspended in l-2ml PBS and blocked with 6% Milk/PBS for a R2 biopanning including a subtraction step. For subtractive selection 6 wells of 96 well NUNC MAXISORP were coated with 0.5pg Chromopure Rabbit IgG (Jackson, 011-000-003) in PBS. For mAb 66- 3-9C, Chromopure Human IgG (Jackson, 009-000-003) was used. Plates were washed and blocked as before. After washing, each Chromopure coated well was incubated with 100 pL of blocked phage at room temperature for one hour with gentle shaking. Unbound phage were transferred to 3-6 wells of 96 well NUNC MAXISORP plate coated with 0.5 pg of monoclonal antibody of interest and pre-blocked with 3% Milk/PBS for the second round of panning. Phage were eluted (R2), and used to infect TGI cells as previously for R1. Finally, R2 bacterial lawn were scraped and glycerol stock were prepared and flash frozen at -80°C.

R2 phagemid DNA was extracted and purified from bacterial glycerol stocks; 100 pL of each bacterial library was thawed and phagemid DNA purified using QIAprep Spin Miniprep kit (QIAGEN, 27104). DNA was eluted in 50 pL H2O.

PCR amplicon DNA containing peptide sequences of interest (9mer or 13mer) was generated using R2 phagemid DNA as template and barcoded primers were used that added DNA encoded barcodes to distinguish libraries generated by panning against different antibodies and that added PI adaptor sequences designed to enable attachment of DNA to beads for emulsion PCR required for Ion Torrent sequencing platform. For 9mer phagemid libraries a single step PCR was utilised. For 13mer phagemid libraries a preparatory PCR was required to incorporate an adaptor sequence. Between PCRs samples were column purified using QiaQuick PCR Purification kit (QIAGEN) to remove primers.

PCR products were separated by gel electrophoresis using low melting point agarose and bands at ~350bp were excised and purified using Nucleospin columns (Machery- Nagel, 740609-250). DNA concentration was quantified using A260 (NanoDrop spectrophotometer) and a Bioanalyser 2100 high sensitivity DNA chip. 100 ng of DNA from each individually barcoded library was pooled. Pooled DNA was purified using magnetic AMPure XP beads (Beckman Coulter, A63880) and DNA concentration quantified using A260 (NanoDrop spectrophotometer).

Pooled barcoded DNA was sequenced via an Ion Torrent PGM service (Macrogen, Inc.) on a 318 chip.

NGS data was processed as described in Naqid et al, Scientific Reports, 2016, 6: 24232-24232.

A two proportion Z-score analysis (Zhang et al, PNAS, 2011) was conducted to identify peptides specifically enriched by mAh of interest. For the 9mer library, each set of peptides enriched by a mAh of interest was compared to those enriched by mAh 11886. Peptides enriched by mAh 11886 were compared to those enriched by human GP -binding antibody 66-3-9C. For the 13mer library, each set of peptides enriched by a target mAh of interest was compared to those enriched by a control non-GP specific rabbit IgG which was panned against the library in parallel to the target GP -binding mAbs. Z-score takes into account the frequency of a peptide sequence in the total number of sequences in the sample, as well as the ratio of the number of copies of a sequence in the sample isolated against the target antibody and number of copies of the same sequence in the sample isolated against the control antibody. Z-score can be used to rank peptide sequences by relative statistical importance and is calculated using the formula: pi — p2 z = pl(l — pi) p2(l — p 2) nl n2

Where nl = number of peptide sequences obtained with the target sample; n2 = number of peptide sequences obtained with the control sample; pi = the number of sequences obtained for a specific peptide against the target sample/nl; p2 = the number of sequences obtained for a specific peptide against the control sample/n2 (Naqid et al, Scientific Reports, 2016).

Peptides were ranked based on Z-score i.e. for specific enrichment in the total reads from Ion torrent sequencing by target mAh of interest. The top 100 peptides were used in all further analysis.

Motifs in the top 100 ranked peptides by z-score were identified using the Multiple EM for Motif Elicitation (MEME) algorithm (Bailey et al, Nucleic Acids Research , 1994). For MEME analysis it was assumed that each motif was expected to occur zero or one time per sequence (zoop). Algorithm was constrained such that motifs occur in a minimum number of ten peptide sequences. Motifs with an E value <0.05 were considered significant.

Peptide motifs were aligned with GP sequences from EBOV (ATY51135), SUDV (YP_138523.1), BDBV (YP_003815435.1) and TAFV (YP_003815426.1) using Clustal Omega algorithm and MegAlignPro software (Version: 11.2.1).

Example 7: Competitive immunofluorescence assay

This assay was used to determine if a GP antibody of interest competed for binding with GP antibodies of known epitope. A panel of antibodies including >1 mAh for each epitope bin was used to confirm competition and place new antibodies of interest in one of three epitope bins; base, glycan cap or receptor binding region.

Antibodies cl3C6, c4G7, c2G4, CA45, 6D6, FVM02 and ADI15878 have epitopes published in the literature by other groups. Antibodies 6541, 66-4-C12, 6660, 6662, 66-3- 9C, 66-3 -2C and 040 are fully human antibodies with epitopes determined via multiple assays as previously described (Rijal, P., et al, Cell Reports, 2019). Antibody 21-D8-5A is an influenza neuraminidase-specific antibody acting as a negative control. Antibodies 6541, 66-4-C12, c2G4 and c4G7 represent different overlapping base epitopes. Antibodies 6660 and 6662 are designated as RBR binding. 66-3-9C, 66-3-2C, 040 and cl3C6 represent different GC epitopes. CA45, 6D6, FVM02, and ADI15878 bind to the conserved fusion loop. It is noted all novel antibodies tested all competed with cl3C6, irrespective of the other mAbs they competed in the panel, hence any apparent competition with cl3C6 is likely not meaningful.

Antibodies were biotinylated using EZ-Link Sulfo-NHS-Biotin kit (Thermo Scientific, 21326). Free biotin was removed using Zeba desalting 7K columns. Biotinylation of mAbs confirmed via dot blot revealed with ExtrAvidin-alkaline phosphatase conjugate (Sigma, E2636) andNBT/BCIP development.

In 96 well flat bottom tissue culture plates, MDCK SIAT-1 cells stably transfected to express EBOV GP were seeded at a density of 3xl0^A5/per well in 100 pL D10 media and incubated for 18 hours, 37°C, 5% CO2. Biotinylated mAbl at 5 pg/mL in PBS and unbiotinylated mAb2 at 50 pg/mL in PBS were mixed in equal volume prior to addition to cells. Each assay plate includes competition controls where mAbl=mAb2 and additionally where mAb2 is a mAb against an irrelevant antigen as a minimum competition control. Cells were washed in PBS to remove D10 media, and 50 pL mix of biotinylated and unbiotinylated mAbs added. Cells were incubated for 1 hr at 4°C. Plates were washed three times with PBS. 50 pL of 2 pg/mL of Streptavidin Alexa Fluor® 647 conjugate (Invitrogen, S21374) was added to each well and cells incubated for 1 hr at 4°C. Plates were washed three times with PBS and cells fixed with 1% formalin/PBS before fluorescence was measured using Clariostar plate reader. Degree of competition was determined by: (X-Minimum binding)/(Maximum binding - Minimum binding), where ‘X’ is binding of the biotinylated mAb in presence of competing mAb, ‘minimum binding’ is the signal from the biotinylated mAb in the presence of self (unbiotinylated) and ‘maximum binding’ is the signal from the biotinylated mAb in presence of a non competing non-GP mAb.

Results are presented in Figures 4 and 5.

Example 8: Binding to thermolysin digested glycoproteins

During infection of a cell, Ebola virus GP is cleaved by cathepsins in the endosome in a process necessary for binding of GP to the human receptor NPC-C1. Thermolysin mimics cathepsin cleavage of GP by removing the glycan cap and mucin-like domains revealing more of the receptor binding region and leaving base epitopes intact. By comparing the binding of mAbs of interest to the GP with and without thermolysin treatment, it can be determined if binding to the GP is dependent on the presence of the glycan cap and mucin-like domain portions of the GP being present.

In 96 well round bottom plates, a minimum of 10^L5 MDCK SIAT-1 cells stably expressing GP from EBOV, SUDV, BDBV or TAFV per well seeded in D10 media (DMEM, Penicillin/Streptomycin 100U each, 2mM Glutamine) and incubated for 18 hours, 37°C, 5% C02.

Cells were washed and incubated in 100 pL 0.25mg/mL thermolysin (Sigma, P1512) in HM buffer (20mM MES/1M HEPES/5M NaCl in dH20) or HM buffer alone for lhr,

37°C, 5% C02. After washing, cells were incubated with 50 uL 10 pg/mL of mAbs of interest and control human mAbs known to bind the base, glycan cap, for lhr, at room temperature with gentle shaking and protected from light. MR78 is a published receptor binding region antibody that cannot bind Ebola virus GP unless the glycan cap domain has been removed (Hashiguchi et al, Cell, 2015); in this assay MR78 directly labelled with Alexa Fluor 647 was used as positive control for digestion of GP by thermolysin. After washing, cells were incubated with 50 uL 1 :400 Goat anti-Rabbit IgG (H+L) Cross- Adsorbed Secondary Antibody Alexa Fluor 647 conjugate (Invitrogen, A21244) (for rabbit mAbs) or Goat anti -Human IgG (H+L) Cross- Adsorbed Secondary Antibody Alexa Fluor 647 conjugate (Invitrogen, A21445) (for control human mAbs), and 5pg/ml of wheat germ agglutinin Alexa Fluor 488 conjugate (Invitrogen, W 11261) for lhr, at room temperature with gentle shaking and protected from light. Cells were washed three times after each incubation with PBS/0.02% sodium azide/0.1% BSA, with plates spun at 500g and supernatant aspirated between washes.

Cells were washed and fixed (PBS/1% formalin/0.1% BSA) before plates were spun to collect all cells at the bottom of plate wells and fluorescence read at both 625-30/680-30 and 488-14/535-30 using a Clariostar plate reader. Gain was adjusted to give a ratio of approximately 1 in wells containing cells that had been stained with both WGA-488 and anti-Rabbit IgG-647 conjugates only (i.e. no primary antibody added). Wells with too few cells as determined by WGA-488 signal were excluded from analysis. AF647 signal normalised to AF488 signal for analysis.

Results are present in Figure 6

Results and discussion 11886 antibody

11886 is a broadly neutralising antibody and can neutralise all three pseudotyped viruses tested (Figure 3). IC50 values are within 2.5 fold of those of CA45 in the same assay against EBOV, SUDV and BDBV GP pseudotypes (Figure 2). This assay suggests that compared to CA45, 11886 neutralisation of the EBOV S-FLU pseudotype is less potent, but neutralisation of SUDV S-FLU pseudotype is more potent. The BDBV S-FLU pseudotype neutralisation profile for both mAbs is highly similar. Neutralisation of the EBOV S-FLU pseudotype in vitro has shown strong though incomplete correlation with protection against EBOV in a mouse challenge model (Rijal et al, Cell Rep, 2019). No comparison has been conducted for neutralisation of the SUDV S-FLU pseudotype, but CA45 (a published broadly protective mAb in small animal models) is protective in SUDV challenge models (Zhao et al, Cell, 2017) and was also neutralising in our SUDV S-FLU in vitro neutralisation assay. 11886 competes for binding to full length EBOV GP expressed on cells with antibodies 6541, c4G7 and c2G4 that bind the base of GP chalice, but not antibodies 66-3- 9C, 66-3 -2C and 040 that bind the glycan cap or antibodies 6660 and 6662 that bind the receptor binding region (Figure 5 A). Unlike antibody 11892 which also competes the base binding antibodies in the epitope binning panel for binding to EBOV GP (Figure 5C), binding of 11886 to GP is not completely independent of the presence of the glycan cap (Figure 6). Binding of 11886 to EBOV GP is reduced to background after digestion of the GP by thermolysin. Binding to SUDV GP is partially reduced after thermolysin digestion. This indicates an intermediate epitope between the glycan cap and chalice base that is not part of the glycan cap, but that is affected by thermolysin digestion. This distinguishes the epitope of this antibody from other base mAbs such as 11892 or 6541 and from true glycan cap antibodies such as 11897 or 040.

For finer mapping of the contacts between 11886 and the GP, 11886 was used to pan two different random combinatorial linear peptide libraries across three independent experiments. Z score analysis was applied to compare sequences enriched by 11886 and a control antibody. The top one hundred sequences specifically enriched by 11886 were analysed for motifs using the MEME tool (Bailey et al, Nucleic Acids Research, 1994) with an arginine-cysteine-arginine (RCR) motif highly represented with similar motifs enriched across experiments and in the independent libraries (Figure 7). Motifs derived from peptides enriched by 11886 predominantly align with a conserved portion of the GP sequences in GP1 (Figure 8 ) that are not in the glycan cap domain, but the exterior of the receptor binding chalice (Figure 10A, B) (GP residues 102-108 and 132-143, with motif of most confidence: R134, C135, R136).

Many of the peptides enriched by 11886 also contain an additional cysteine with spacing consistent with formation of an intra-peptide disulphide bond and a non-linear peptide. This structural feature of the peptides is consistent with a turn structure seen in the GP in the middle of the arginine-cysteine-arginine (RCR) motif (Figure 9).

In the context of a previously published electron microscopy structures of EBOV GP with the glycan cap present (Figure 10A, B; Protein data bank (PDB): 5KEL), and with the glycan cap domain removed (Figure IOC, D; PDB: 6MAM) the residues in the GP that align with peptide phage display derived motifs are surface exposed and not part of the glycan cap. However, these residues sit very close to the surface that is affected by removal of the glycan cap which is consistent with sensitivity to thermolysin cleavage. The location of the motifs on the outside of the receptor binding region chalice but not the glycan cap is consistent with the competition data showing competition of base binding antibodies, but not those that bind the glycan cap.

11886 does not strongly compete known fusion loop antibodies tested, especially those that are known to bind the fusion loop tip; the assay does indicate some, although not strong, competition with ADI-15878 (Figure 5B). ADI-15878 has a published epitope spanning the paddle of fusion loop and contacts the GP predominantly in GP2, and the residues identified as involved in binding of this mAh to GP do not include those identified for 11886 by peptide phage display (Wee et al, Cell, 2017; West et al, mBio, 2018). The partial competition seen may be due to the angle that these mAbs bind rather than overlapping binding footprint on the GP.

Although not tested experimentally in the competition assay, the epitope of 11886 as defined by the peptide mapping is distinct form epitopes in the literature for other broadly protective mAbs.

The location of the 11886 binding region is proximal to, but not the same as, the binding region of fab ADI-15946 (Figure 10D, PDB:6MAM). The contacts made by ADI- 15946 involve the regions 71-77 (GP1), 251-303) (GC) and 508-514 (GP2) with a K510E escape mutant sufficient for complete escape of the virus from this antibody (Wee et al, Cell, 2017; West et al, Nature Structural & Molecular Biology, 2019).

A cocktail of afucosylated versions of ADI-23774 (a derivative of ADI-15946) and ADI-15878 named MBP134AF has been shown to be protective against EBOV, SUDV and BDBV challenge in non-human primates (Bomholdt et al, Cell Rep, 2019). Antibody 11886 represents a broadly neutralising antibody that targets an epitope distinct to those in MBP134AF and thus a potential additional component to such cocktails as their design continues to develop.

Antibody FVM04 which has shown to be protective in EBOV and SUDV animal models and was able to partially neutralise BDBV in vitro binds to the crest of the chalice at the receptor binding region (Howell et al, Cell Rep, 2016). The key residues for binding of this mAh identified by alanine scanning (K115, D117, and G118) (Figure 10E) are not those identified for 18866 binding and sit higher up the chalice crest than the RCR motif. Antibody m2 ID 10 is a broadly reactive, though non-neutralising mAb, that binds to GP from EBOV, SUDV (although with lower affinity) BDBV, RESTV as well as a related filovirus, Marburg virus (Holtsberg et al, J Virol, 2016). Like 11886 it also has an epitope on the outside of the receptor binding chalice; however, this antibody shows improved binding after removal of the glycan cap and its epitope constitutes residues 81-90 placing its epitope on the opposite side of the GP1 monomer to the RCR binding motif of 11886 (Figure 10E).

Therefore antibody 11886 represents a novel broadly-reactive broadly-neutralising antibody that has an epitope distinct from those already published. The current data suggest the epitope of this antibody sits on the chalice between the crest epitope represented by FVM04, and above the fusion loop antibodies and base epitope of ADI- 15946.

Example 9: In silico humanisation of sequences

Antibodies were humanised in silico by grafting the CDRs from the rabbit antibody V-regions onto human germline antibody V-region frameworks. In order to recover the activity of the antibody, a number of framework residues from the rabbit V-regions were also retained in the humanised sequences. These residues were selected using the protocol outlined by Adair et al. (1991) (Humanised antibodies. WO91/09967). The CDRs grafted from the donor to the acceptor sequence are as defined by Rabat (Rabat et al., 1987), with the exception of CDRH1 where the combined Chothia/Rabat definition is used (see Adair et al., 1991 Humanised antibodies. WO91/09967). Commonly the VH genes of rabbit antibodies are shorter than the selected human VH acceptor genes. When aligned with the human acceptor sequences, framework 1 of the VH regions of rabbit antibodies typically lack the N-terminal residue, which is retained in the humanised antibody. Framework 3 of the rabbit antibody VH regions also typically lack one or two residues (75, or 75 and 76) in the loop between beta sheet strands D and E: in the humanised antibodies the gap is filled with the corresponding residues from the selected human acceptor sequence.

Antibody 11886

Human V-region IGRV1-5 plus JR4 J-region (IMGT, http://www.imgt.org/) was chosen as the acceptor for antibody 11886 light chain CDRs. In addition to the CDRs, one or more of the following framework residues from the 11886 VK gene (donor residue) may be retained at positions 2 and 3 (Rabat numbering): Valine (V2) and Valine (V3), respectively. In some cases, CDRL3 may be mutated to remove a Cysteine residue (CDRL3 variants; SEQ ID NOs: 4 and 5).

Human V-region IGHV3-66 plus JH5 J-region (IMGT, http://www.imgt.org/) was chosen as an acceptor for the heavy chain CDRs of antibody 11886. In addition to the CDRs, one or more of the following framework residues from the 11886 VH gene (donor residues) may be retained at positions 23, 24, 48, 49, 71, 73 and 78 (Rabat numbering): Isoleucine (123), Valine (V24), Isoleucine (148), Glycine (G49), Lysine (R71), Alanine (A73) and Valine (V78), respectively. In some cases, CDRH3 may be mutated to modify a potential Aspartic Acid-Proline hydrolysis site (CDRH3 variants; SEQ ID NOs: 9 and 10).

Antibody 11897

Human V-region IGRV1D-13 plus JR4 J-region (IMGT, http://www.imgt.org/) was chosen as the acceptor for antibody 11897 light chain CDRs. In addition to the CDRs, one or more of the following framework residues from the 11897 VR gene (donor residue) may be retained at positions 2, 3 and 70 (Rabat numbering): Valine (V2), Valine (V3) and Glutamine (Q70), respectively.

Human V-region IGHV3-30-3 plus JH4 J-region (IMGT, http://www.imgt.org/) was chosen as an acceptor for the heavy chain CDRs of antibody 11897. In addition to the CDRs, one or more of the following framework residues from the 11897 VH gene (donor residues) may be retained at positions 24, 47, 48, 49, 73 and 78 (Rabat numbering): Valine (V24), Tyrosine (Y47), Isoleucine (148), Glycine (G49), Serine (S73) and Valine (V78), respectively. The Glutamine residue at position 1 of the human framework was replaced with Glutamic acid (El) to afford the expression and purification of a homogeneous product: the conversion of Glutamine to pyroGlutamate at the N-terminus of antibodies and antibody fragments is widely reported. In some cases, CDRH2 may be mutated to remove a potential N-linked glycosylation site (CDRH2 variants; SEQ ID NOs: 39-41).

Antibody 11878

Human V-region IGRV1-5 plus JR4 J-region (IMGT, http://www.imgt.org/) was chosen as the acceptor for antibody 11878 light chain CDRs. In addition to the CDRs, one or more of the following framework residues from the 11878 VK gene (donor residue) may be retained at positions 2, 3 and 38 (Kabat numbering): Valine (V2), Valine (V3) and Leucine (L38), respectively.

Human V-region IGHV3-66 plus JH4 J-region (IMGT, http://www.imgt.org/) was chosen as an acceptor for the heavy chain CDRs of antibody 11878. In addition to the CDRs, one or more of the following framework residues from the 11878 VH gene (donor residues) may be retained at positions 24, 48, 49, 71, 73 and 78 (Kabat numbering): Valine (V24), Isoleucine (148), Glycine (G49), Lysine (K71), Serine (S73) and Valine (V78), respectively.

Antibody 11883

Human V-region IGKV1D-13 plus JK4 J-region (IMGT, http://www.imgt.org/) was chosen as the acceptor for antibody 11883 light chain CDRs. In addition to the CDRs, one or more of the following framework residues from the 11883 VK gene (donor residue) may be retained at positions 2, 3 and 71 (Kabat numbering): Valine (V2), Valine (V3) and Tyrosine (Y71), respectively.

Human V-region IGHV3-72 plus JH5 J-region (IMGT, http://www.imgt.org/) was chosen as an acceptor for the heavy chain CDRs of antibody 11883. In addition to the CDRs, one or more of the following framework residues from the 11883 VH gene (donor residues) may be retained at positions 48, 49, 71, 73 and 78 (Kabat numbering): Isoleucine (148), Alanine (A49), Lysine (K71), Serine (S73) and Valine (V78), respectively. In some cases, CDRHl and CDRH2 may be mutated to remove Cysteine residues (CDRHl variants and CDRH2 variants, respectively; SEQ ID NOs: 68, 69, 71 and 72).

Antibody 11889

Human V-region IGKV1D-13 plus JK4 J-region (IMGT, http://www.imgt.org/) was chosen as the acceptor for antibody 11889 light chain CDRs. In addition to the CDRs, one or more of the following framework residues from the 11889 VK gene (donor residue) may be retained at positions 2, 3 and 66 (Kabat numbering): Valine (V2), Valine (V3) and Arginine (R66), respectively.

Human V-region IGHV3-23 plus JH4 J-region (IMGT, http://www.imgt.org/) was chosen as an acceptor for the heavy chain CDRs of antibody 11889. In addition to the CDRs, one or more of the following framework residues from the 11889 VH gene (donor residues) may be retained at positions 48, 49, 71, 73, 76, 78 and 94 (Rabat numbering): Isoleucine (148), Alanine (A49), Lysine (K71), Serine (S73), Threonine (T76), Valine (V78) and Threonine (T94), respectively. In some cases, CDRH1 and CDRH2 may be mutated to remove Cysteine residues (CDRH1 variants and CDRH2 variants, respectively; SEQ ID NOs: 87 and 89).

Antibody 11892

Human V-region IGKV1-12 plus JK4 J-region (IMGT, http://www.imgt.org/) was chosen as the acceptor for antibody 11892 light chain CDRs. In addition to the CDRs, one or more of the following framework residues from the 11892 VK gene (donor residue) may be retained at positions 2 and 3 (Rabat numbering): Valine (V2) and Valine (V3), respectively. In some cases, CDRL3 may be mutated to remove a Cysteine residue (CDRL3 variants; SEQ ID NOs: 101 and 102).

Human V-region IGHV3-66 plus JH5 J-region (IMGT, http://www.imgt.org/) was chosen as an acceptor for the heavy chain CDRs of antibody 11892. In addition to the CDRs, one or more of the following framework residues from the 11892 VH gene (donor residues) may be retained at positions 24, 48, 49, 71, 73, 76 and 78 (Rabat numbering): Valine (V24), Isoleucine (148), Glycine (G49), Lysine (R71), Serine (S73), Threonine (T76) and Valine (V78), respectively. In some cases, CDRH3 may be mutated to modify a potential Aspartic Acid-Proline hydrolysis site (CDRH3 variants; SEQ ID NOs: 106 and 107).

Antibody 11881

Human V-region IGRV1-12 plus JR4 J-region (IMGT, http://www.imgt.org/) was chosen as the acceptor for antibody 11881 light chain CDRs. In addition to the CDRs, one or more of the following framework residues from the 11881 VR gene (donor residue) may be retained at positions 2 and 3 (Rabat numbering): Valine (V2) and Valine (V3), respectively. In some cases, CDRL3 may be mutated to remove a Cysteine residue (CDRL3 variants; SEQ ID NOs: 121 and 122).

Human V-region IGHV3-66 plus JH5 J-region (IMGT, http://www.imgt.org/) was chosen as an acceptor for the heavy chain CDRs of antibody 11881. In addition to the CDRs, one or more of the following framework residues from the 11881 VH gene (donor residues) may be retained at positions 24, 48, 49, 71, 73, 76 and 78 (Rabat numbering): Valine (V24), Isoleucine (148), Glycine (G49), Lysine (K71), Serine (S73), Threonine (T76) and Valine (V78), respectively. In some cases, CDRH3 may be mutated to modify a potential Aspartic Acid-Proline hydrolysis site (CDRH3 variants; SEQ ID NOs: 126 and 127).

Materials

Sequence listing

SEQ ID NO: 1 - 11886 CDRL1

QASQSIGSNLA

SEQ ID NO: 2 - 11886 CDRL2

AASTLAS

SEQ ID NO: 3 - 11886 CDRL3

QCTYYDSSYVYNN

SEQ ID NO: 4 - 11886 CDRL3 variant

QQTYYDSSYVYNN

SEQ ID NO: 5 - 11886 CDRL3 variant

QSTYYDSSYVYNN

SEQ ID NO: 6 - 11886 CDRH1

GFSLSNYYMS

SEQ ID NO: 7 - 11886 CDRH2

VIGWSGTSSYASWAKG

SEQ ID NO: 8 - 11886 CDRH3

VLYIGGGFNYYDAFDP

SEQ ID NO: 9 - 11886 CDRH3 variant

VLYIGGGFNYYDAFEP

SEQ ID NO: 10 - 11886 CDRH3 variant

VLYIGGGFNYYDAFNP

SEQ ID NO: 11 - Rabbit Ab 11886 VL region DW MTQTPASVSEPVGGTVTIKCQASQSIGSNLAWYQQKPGQPPKLLIYAASTLASGVPSR FKGSGSGTEFTLTISDLECADAATYYCQCTYYDSSYVYNNFGGGTEW VK

SEQ ID NO: 12 - Rabbit Ab 11886 VL region Gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagt caeca tcaagtgccaggccagtcagagtattggtagtaatttagcctggtatcagcagaaaccagg gcagcctcccaagctcctgatctatgctgcatccactctggcatctggggtcccatcgcgg ttcaaaggcagtggatctgggacagagttcactctcaccatcagcgacctggagtgtgccg atgctgccacttactactgtcaatgtacttattatgatagtagttatgtttataataattt tggcggagggaccgaggtggtggtcaaa

SEQ ID NO: 13 - Rabbit Ab 11886 VH region

QSVEESGGRLVTPGTPLTLTCIVSGFSLSNYYMSWVRQVPGKGLEWIGVIGWSGTSSYASW

AKGRFTISKTASTTVDLKITSPTTEDTATYFCARVLYIGGGFNYYDAFDPWGPGTLVTVSS

SEQ ID NO: 14 - Rabbit Ab 11886 VH region

Cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacct gcatagtctctggattctccctcagtaactactatatgagctgggtccgccaggttccagg gaaggggctggaatggatcggagtcattggttggagtggtacctcatcctacgcgagctgg gcgaaaggccgattcaccatctccaaaaccgcgtcgaccacggtggatctgaaaat caeca gtccgacaaccgaggacacggccacctatttctgtgccagagtcctatatattggtggtgg ttttaattactatgatgcttttgatccctggggcccaggcaccctggtcaccgtctcgagc

SEQ ID NO: 15 - 11886 humanised VL region

DVVMTQSPSTLSASVGDRVTITCQASQS IGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQCTYYDSSYVYNNFGGGTKVEIK

SEQ ID NO: 16 - 11886 humanised VL region with variant CDR3

DVVMTQSPSTLSASVGDRVTITCQASQS IGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQQTYYDSSYVYNNFGGGTKVEIK

SEQ ID NO: 17 - 11886 humanised VL region with variant CDR3

DVVMTQSPSTLSASVGDRVTITCQASQS IGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQSTYYDSSYVYNNFGGGTKVEIK

SEQ ID NO: 18 - 11886 humanised VH region

EVQLVESGGGLVQPGGSLRLSCIVSGFSLSNYYMSWVRQAPGKGLEWIGVIGWSGTSSYAS

WAKGRFTISKDASKNTVYLQMNSLRAEDTAVYYCARVLYIGGGFNYYDAFDPWGQGTLVTV

SS

SEQ ID NO: 19 - 11886 humanised VH region with variant CDR3

EVQLVESGGGLVQPGGSLRLSCIVSGFSLSNYYMSWVRQAPGKGLEWIGVIGWSGTSSYAS

WAKGRFTISKDASKNTVYLQMNSLRAEDTAVYYCARVLYIGGGFNYYDAFEPWGQGTLVTV

SS SEQ ID NO: 20 - 11886 humanised VH region with variant CDR3

EVQLVESGGGLVQPGGSLRLSCIVSGFSLSNYYMSWVRQAPGKGLEWIGVIGWSGTSSYAS

WAKGRFTISKDASKNTVYLQMNSLRAEDTAVYYCARVLYIGGGFNYYDAFNPWGQGTLVTV

SS

SEQ ID NO: 21 - Bundibugyo Ebola virus glycoprotein sequence (NBCI reference YP_003815435.1)

MVTSGILQLPRERFRKTSFFVWVIILFHKVFPIPLGVVHNNTLQVSDIDKLVCRDKLSSTS QLKSVGLNLEGNGVATDVPTATKRWGFRAGVPPKW NYEAGEWAENCYNLDIKKADGSECL PEAPEGVRGFPRCRYVHKVSGTGPCPEGYAFHKEGAFFLYDRLASTI IYRSTTFSEGW AF LILPETKKDFFQSPPLHEPANMTTDPSSYYHTVTLNYVADNFGTNMTNFLFQVDHLTYVQL EPRFTPQFLVQLNETIYTNGRRSNTTGTLIWKVNPTVDTGVGEWAFWENKKNFTKTLSSEE LSVIFVPRAQDPGSNQKTKVTPTSFANNQTSKNHEDLVPEDPASW QVRDLQRENTVPTPP PDTVPTTLIPDTMEEQTTSHYEPPNISRNHQERNNTAHPETLANNPPDNTTPSTPPQDGER TSSHTTPSPRPVPTSTIHPTTRETHIPTTMTTSHDTDSNRPNPIDISESTEPGPLTNTTRG AANLLTGSRRTRREITLRTQAKCNPNLHYWTTQDEGAAIGLAWIPYFGPAAEGI YTEGIMH NQNGLICGLRQLANETTQALQLFLRATTELRTFS ILNRKAIDFLLQRWGGTCHILGPDCCI EPHDWTKNITDKIDQIIHDFIDKPLPDQTDNDNWWTGWRQWVPAGIGITGVI IAVIALLCI CKFLL

SEQ ID NO: 22 - Zaire Ebola virus glycoprotein sequence (ATY51135.1)

MGVTGILQLPRDRFKRTSFFLWVIILFQRTFS IPLGVIHNSTLQVSDVDKLVCRDKLSSTN QLRSVGLNLEGNGVATDVPSATKRWGFRSGVPPKW NYEAGEWAENCYNLEIKKPDGSECL PAAPDGIRGFPRCRYVHKVSGTGPCAGDFAFHKEGAFFLYDRLASTVI YRGTTFAEGW AF LILPQAKKDFFSSHPLREPVNATEDPSSGYYSTTIRYQATGFGTNETEYLFEVDNLTYVQL ESRFTPQFLLQLNETIYASGKRSNTTGKLIWKVNPEIDTTIGEWAFWETKKNLTRKIRSEE LSFTAVSNGPKNISGQSPARTSSDPETNTTNEDHKIMASENSSAMVQVHSQGRKAAVSHLT TLATISTSPQPPTTKTGPDNSTHNTPVYKLDISEATQVGQHHRRADNDSTASDTPPATTAA GPLKAENTNTSKSADSLDLATTTSPQNYSETAGNNNTHHQDTGEESASSGKLGLITNTIAG VAGLITGGRRTRREVIVNAQPKCNPNLHYWTTQDEGAAIGLAWIPYFGPAAEGI YIEGLMH NQDGLICGLRQLANETTQALQLFLRATTELRTFS ILNRKAIDFLLQRWGGTCHILGPDCCI EPHDWTKNITDKIDQIIHDFVDKTLPDQGDNDNWWTGWRQWIPAGIGVTGVI IAVIALFCI CKFVF

SEQ ID NO: 23 - Sudan Ebola virus glycoprotein sequence (NCBI Reference Sequence: YP_138523.1)

MGGLSLLQLPRDKFRKSSFFVWVIILFQKAFSMPLGW TNSTLEVTEIDQLVCKDHLASTD QLKSVGLNLEGSGVSTDIPSATKRWGFRSGVPPKW SYEAGEWAENCYNLEIKKPDGSECL PPPPDGVRGFPRCRYVHKAQGTGPCPGDYAFHKDGAFFLYDRLASTVI YRGVNFAEGVIAF LILAKPKETFLQSPPIREAVNYTENTSSYYATSYLEYEIENFGAQHSTTLFKIDNNTFVRL DRPHTPQFLFQLNDTIHLHQQLSNTTGRLIWTLDANINADIGEWAFWENKKNLSEQLRGEE LSFEALSLNETEDDDAASSRITKGRISDRATRKYSDLVPKNSPGMVPLHIPEGETTLPSQN STEGRRVGVNTQETITETAATIIGTNGNHMQISTIGIRPSSSQIPSSSPTTAPSPEAQTPT THTSGPSVMATEEPTTPPGSSPGPTTEAPTLTTPENITTAVKTVLPQESTSNGLITSTVTG ILGSLGLRKRSRRQTNTKATGKCNPNLHYWTAQEQHNAAGIAWIPYFGPGAEGI YTEGLMH NQNALVCGLRQLANETTQALQLFLRATTELRTYTILNRKAIDFLLRRWGGTCRILGPDCCI EPHDWTKNITDKINQIIHDFIDNPLPNQDNDDNWWTGWRQWIPAGIGITGI IIAIIALLCV CKLLC

SEQ ID NO: 24 - Tai Forest Ebola virus glycoprotein sequence (NCBI Reference Sequence: YP 003815426.1)

MGASGILQLPRERFRKTSFFVWVI ILFHKVFSIPLGVVHNNTLQVSDIDKFVCRDKLSSTS QLKSVGLNLEGNGVATDVPTATKRWGFRAGVPPKW NCEAGEWAENCYNLAIKKVDGSECL PEAPEGVRDFPRCRYVHKVSGTGPCPGGLAFHKEGAFFLYDRLASTI IYRGTTFAEGVIAF LILPKARKDFFQSPPLHEPANMTTDPSSYYHTTTINYVVDNFGTNTTEFLFQVDHLTYVQL EARFTPQFLVLLNETIYSDNRRSNTTGKLIWKINPTVDTSMGEWAFWENKKNFTKTLSSEE LSFVPVPETQNQVLDTTATVSPP ISAHNHAAEDHKELVSEDSTPW QMQNIKGKDTMPTTV TGVPTTTPSPFPINARNTDHTKSFIGLEGPQEDHSTTQPAKTTSQPTNSTESTTLNPTSEP SSRGTGPSSPTVPNTTESHAELGKTTPTTLPEQHTAASAIPRAVHPDELSGPGFLTNTIRG VTNLLTGSRRKRRDVTPNTQPKCNPNLHYWTALDEGAAIGLAWIPYFGPAAEGI YTEGIME NQNGLICGLRQLANETTQALQLFLRATTELRTFS ILNRKAIDFLLQRWGGTCHILGPDCCI EPQDWTKNITDKIDQIIHDFVDNNLPNQNDGSNWWTGWKQWVPAGIGITGVI IAIIALLCI CKEML

SEQ ID NO: 25 Glycoprotein consensus sequence

PRCRYVHK

SEQ ID NO: 26 9mer motif 1 RCRSW C

SEQ ID NO: 27 9mer motif 2

KFCQTCQ

SEQ ID NO: 28 9mer3 motif 3

YLRRSRG SEQ ID NO: 29 9mer2 motif 1.1

GKRCRSVDC

SEQ ID NO: 30 9mer2 motif 1.2

GKRCRGVDC SEQ ID NO: 31 - 9mer2 motif 2

GRCRSSV SEQ ID NO: 32 - 13mer motif 1

PRCRSII

SEQ ID NO: 33 - 13mer motif 2

GGWCGRG

SEQ ID NO: 34 - 11897 CDRL1 QASQNVYGLLA

SEQ ID NO: 35 - 11897 CDRL2

SASTLAS

SEQ ID NO: 36 - 11897 CDRL3

QRYYYSSGTTETT SEQ ID NO: 37 - 11897 CDRH1

GFDLSSYAM

SEQ ID NO: 38 - 11897 CDRH2

IINYSGNRYYASWAKG

SEQ ID NO: 39 - 11897 CDRH2 variant I IDYSGNRYYASWAKG

SEQ ID NO: 40 - 11897 CDRH2 variant

IISYSGNRYYASWAKG

SEQ ID NO: 41 - 11897 CDRH2 variant IIQYSGNRYYASWAKG SEQ ID NO: 42 - 11897 CDRH3

GGYDDYGYVSYFDI SEQ ID NO: 43 - 11897 rabbit VL (amino acid)

AW LTQTPASVSAPVGGTVTINCQASQNVYGLLAWYQQKPGQPPKLLIYSASTLASGVPSR

FKGSGSGTQFTLTISDLECADAATYYCQRYYYSSGTTETTFGGGTEW VK

SEQ ID NO: 44 - 11897 rabbit VL (nucleic acid) gccgtcgtgttgacccagactccagcctctgtgtctgcacctgtgggaggcacagt caeca tcaattgccaggccagtcagaatgtttatggtttattggcctggtatcaacagaaaccagg gcagcctcccaagctcctgatctattctgcatccactctggcatctggggtcccatcgcga ttcaaaggcagtggatctgggacacagttcactctcaccatcagcgacctggagtgtgccg atgctgccacttactactgtcaaaggtattattatagtagtggtactactgagactacttt tggcggagggaccgaggtggtggtcaaa

SEQ ID NO: 45 - 11897 rabbit VH (amino acid)

QSVEESGGRLVTPGTPLTLTCTVSGFDLSSYAMGWVRQAPGKGLEYIGI INYSGNRYYASW AKGRFTISRTSTTVDLSMTSLTTEDTATYFCARGGYDDYGYVSYFDIWGPGTLVTVSS

SEQ ID NO: 46 - 11897 rabbit VH (nucleic acid) cagtcggtggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacct gcacagtctctgggttcgacctcagtagctatgcaatgggctgggtccgccaggctccagg gaaggggctggaatacatcggaatcattaattatagtggtaacagatattacgcgagctgg gcgaaaggccgattcaccatctccagaacctcgaccacggtggatctgtcaatgaccagtc tgacaaccgaggacacggccacctatttctgtgccagagggggttatgatgattatggtta tgtgtcctactttgacatctggggcccaggcaccctggtcaccgtctcgagc

SEQ ID NO: 47 - humanised 11897 VL

AVVLTQSPSSLSASVGDRVTITCQASQNVYGLLAWYQQKPGKAPKLLI YSASTLASGVPSR FSGSGSGTQFTLTISSLQPEDFATYYCQRYYYSSGTTETTFGGGTKVEIK

SEQ ID NO: 48 - humanised 11897 VH

EVQLVESGGGW QPGRSLRLSCAVSGFDLSSYAMGWVRQAPGKGLEYIGIINYSGNRYYAS WAKGRFTISRDSSKNTVYLQMNSLRAEDTAVYYCARGGYDDYGYVSYFDIWGQGTLVTVSS

SEQ ID NO: 49 - humanised 11897 VH (CDRH2 variant)

EVQLVESGGGW QPGRSLRLSCAVSGFDLSSYAMGWVRQAPGKGLEYIG11DYSGNRYYAS WAKGRFTISRDSSKNTVYLQMNSLRAEDTAVYYCARGGYDDYGYVSYFDIWGQGTLVTVSS

SEQ ID NO: 50 - humanised 11897 VH (CDRH2 variant)

EVQLVESGGGW QPGRSLRLSCAVSGFDLSSYAMGWVRQAPGKGLEYIGIISYSGNRYYAS WAKGRFTISRDSSKNTVYLQMNSLRAEDTAVYYCARGGYDDYGYVSYFDIWGQGTLVTVSS SEQ ID NO: 51 - humanised 11897 VH (CDRH2 variant)

EVQLVESGGGW QPGRSLRLSCAVSGFDLSSYAMGWVRQAPGKGLEYIG11QYSGNRYYAS WAKGRFTISRDSSKNTVYLQMNSLRAEDTAVYYCARGGYDDYGYVSYFDIWGQGTLVTVSS

SEQ ID NO: 52 - 11878 CDRL1

QASENIDNLLA

SEQ ID NO: 53 - 11878 CDRL2

PASTLAS

SEQ ID NO: 54 - 11878 CDRL3

QSNYYGFYYGMT

SEQ ID NO: 55 - 11878 CDRH1

GFSLSSNDMN

SEQ ID NO: 56 - 11878 CDRH2

HIWSGGSTYYPSWARG

SEQ ID NO: 57 - 11878 CDRH3

GPVSDI

SEQ ID NO: 58 - 11878 rabbit VL (amino acid)

DW MTQTASPVSAPVGGTVTIKCQASENIDNLLAWYQLKPGQPPKLLIYRASTLASGVPSR FKGSGSGTEFTLTISGVQCDDAATYYCQSNYYGFYYGMTFGGGTEW VK

SEQ ID NO: 59 - 11878 rabbit VL (nucleic acid) gatgttgtgatgacccagactgcatcccccgtgtctgcacctgtgggaggcacagt caeca tcaagtgccaggccagtgagaacattgataacttattggcctggtatcagctgaaaccagg gcagcctcccaagctcctgatctacagggcatccactctggcatctggggtcccatcgcgg ttcaaaggcagtggatctgggacagagttcactctcaccatcagcggcgtgcagtgtgacg atgctgccacttactactgtcaaagcaattattatggcttttattatggtatgactttcgg cggagggaccgaggtggtggtcaaa

SEQ ID NO: 60 - 11878 rabbit VH (amino acid)

QSVEESGGRLVTPGTPLTLTCTVSGFSLSSNDMNWVRQAPGKGLEWIGHIWSGGSTYYPSW

ARGRFTISKTSTTVDLKITSPTSEDTATYFCARGPVSDIWGPGTLVTVSS SEQ ID NO: 61 11878 rabbit VH (nucleic acid) cagtcggtggaggagtcggggggtcgcctggtcacgcctgggacacccctgacactcacct gcacagtctctggattctccctcagcagcaacgacatgaactgggtccgccaggctccagg gaaggggctggagtggatcggacacatttggagtggtggtagtacatactacccgagctgg gcgagaggccgattcaccatctccaaaacctcgaccacggtggatctgaaaatcaccagtc cgacaagcgaggacacggccacctatttctgtgccagagggcctgttagtgacatctgggg cccaggcaccctggtcaccgtctcgagc

SEQ ID NO: 62 - humanised 11878 VL

DVVMTQSPSTLSASVGDRVTITCQASENIDNLLAWYQLKPGKAPKLLI YRASTLASGVPSR FSGSGSGTEFTLTISSLQPDDFATYYCQSNYYGFYYGMTFGGGTKVEIK

SEQ ID NO: 63 - humanised 11878 VH

EVQLVESGGGLVQPGGSLRLSCAVSGFSLSSNDMNWVRQAPGKGLEWIGHIWSGGSTYYPS

WARGRFTISKDSSKNTVYLQMNSLRAEDTAVYYCARGPVSDIWGQGTLVTVSS

SEQ ID NO: 64 - 11883 CDRL1

QASQNIYSNLA

SEQ ID NO: 65 - 11883 CDRL2

SASTLAS

SEQ ID NO: 66 - 11883 CDRL3

QRYYYLSGSADNT

SEQ ID NO: 67 - 11883 CDRH1

GFSFSSNCWRC

SEQ ID NO: 68 - 11883 CDRH1 variant

GFSFSSNSWRC

SEQ ID NO: 69 - 11883 CDRH1 variant

GFSFSSNCWRS

SEQ ID NO: 70 - 11883 CDRH2

CVCAGRSGGTTYYASWAKG

SEQ ID NO: 71 - 11883 CDRH2 variant CVSAGRSGGTTYYASWAKG

SEQ ID NO: 72 - 11883 CDRH2 variant

SVCAGRSGGTTYYASWAKG

SEQ ID NO: 73 - 11883 CDRH3

AGYDDYGDASFFNL

SEQ ID NO: 74 - 11883 rabbit VL (amino acid)

AW LTQTASPVSTPVGGTVTIKCQASQNIYSNLAWYQQKPGQPPKLLIYSASTLASGVPSR FKGSGSGTEYTLTISDLECADAATYYCQRYYYLSGSADNTFGGGTEW VK

SEQ ID NO: 75 - 11883 rabbit VL (nucleic acid) gccgtcgtgctgacccagactgcatcccccgtgtctacacctgtgggaggcacagt caeca tcaagtgccaggccagtcagaacatttacagtaatttagcctggtatcagcagaaaccagg gcagcctcccaagctcctgatctattctgcatccactctggcatctggggtcccatcgcgg ttcaaaggcagtggatctgggacagagtacactctcaccatcagcgacctggagtgtgccg atgctgccacttactactgtcaaaggtattattatcttagtggtagtgctgataatacttt cggcggagggaccgaggtggtggtcaaa

SEQ ID NO: 76 - 11883 rabbit VH (amino acid)

QEQLVESGGGLVQPEGSLTLTCTASGFSFSSNCWRCWVRQAPGKGLEWIACVCAGRSGGTT

YYASWAKGRFTISKTSSPTVTLQMTSLTAADTATYFCARAGYDDYGDASFFNLWGPGTLVT

VSS

SEQ ID NO: 77 - 11883 rabbit VH (nucleic acid) caggagcagctggtggagtccgggggaggcctggtccagcctgagggatccctgacactca cctgcacagcctctggattctccttcagtagcaactgctggagatgctgggtccgccaggc tccagggaaggggctggagtggatcgcatgcgtttgtgctggtaggagtggtggtaccact tactacgcgagctgggcgaaaggccgattcaccatctccaaaacctcgtcgcccacggtga ctcttcaaatgaccagtctgacagccgcggacacggccacctatttctgtgcgagagctgg ttatgatgattatggtgacgcttccttctttaacttgtggggcccaggcaccctggtcacc gtetegage

SEQ ID NO: 78 - humanised 11883 VL

AVVLTQSPSSLSASVGDRVTITCQASQNIYSNLAWYQQKPGKAPKLLI YSASTLASGVPSR FSGSGSGTDYTLTISSLQPEDFATYYCQRYYYLSGSADNTFGGGTKVEIK

SEQ ID NO: 79 - humanised 11883 VH EVQLVESGGGLVQPGGSLRLSCAASGFSFSSNCWRCWVRQAPGKGLEWIACVCAGRSGGTT YYASWAKGRFTISKDSSKNSVYLQMNSLKTEDTAVYYCARAGYDDYGDAS FFNLWGQGTLV TVSS SEQ ID NO: 80 - humanised 11883 VH (CDRH1 and 2 variant)

EVQLVESGGGLVQPGGSLRLSCAASGFSFSSNSWRCWVRQAPGKGLEWIACVSAGRSGGTT YYASWAKGRFTISKDSSKNSVYLQMNSLKTEDTAVYYCARAGYDDYGDAS FFNLWGQGTLV TVSS

SEQ ID NO: 81 - humanised 11883 VH (CDRH1 and 2 variant)

EVQLVESGGGLVQPGGSLRLSCAASGFSFSSNCWRSWVRQAPGKGLEWIASVCAGRSGGTT YYASWAKGRFTISKDSSKNSVYLQMNSLKTEDTAVYYCARAGYDDYGDAS FFNLWGQGTLV TVSS

SEQ ID NO: 82 - humanised 11883 VH (CDRH1 and 2 variant)

EVQLVESGGGLVQPGGSLRLSCAASGFSFSSNSWRSWVRQAPGKGLEWIASVSAGRSGGTT YYASWAKGRFTISKDSSKNSVYLQMNSLKTEDTAVYYCARAGYDDYGDAS FFNLWGQGTLV TVSS

SEQ ID NO: 83 - 11889 CDRL1

QASQNIYSDLA

SEQ ID NO: 84 - 11889 CDRL2

DTSNLAS

SEQ ID NO: 85 - 11889 CDRL3

QAYYYSSSSGDTT

SEQ ID NO: 86 - 11889 CDRH1

GFSFSSSYWIC

SEQ ID NO: 87 - 11889 CDRH1 variant

GFSFSSSYWIS

SEQ ID NO: 88 - 11889 CDRH2

ClYAGSSGSTYYASWAKG

SEQ ID NO: 89 11889 CDRH2 variant SIYAGSSGSTYYASWAKG

SEQ ID NO: 90 - 11889 CDRH3

AYAVSAPFGYTLFRYFEL

SEQ ID NO: 91 - 11889 rabbit VL (amino acid)

AW LTQTASPVSAPVGGTVTIKCQASQNIYSDLAWYQQKPGQPPKLLIYDTSNLASGVSSR FKGSRSGTEFTLTISDLECADAATYYCQAYYYSSSSGDTTFGGGTEW VK

SEQ ID NO: 92 - 11889 rabbit VL (nucleic acid) gccgtcgtgctgacccagactgcatcccccgtgtctgcacctgtgggaggcacagt caeca tcaagtgccaggccagtcagaacatttacagcgatttagcctggtatcagcagaaaccagg gcagcctcccaagctcctgatctatgatacatccaatctggcatctggggtctcatcgcgg ttcaaaggcagtagatctgggacagagttcactctcaccatcagcgacctggagtgtgccg atgctgccacttactactgtcaagcctattattatagtagtagtagtggtgatactacttt cggcggagggaccgaggtggtggtcaaa

SEQ ID NO: 93 - 11889 rabbit VH (amino acid)

QSLEESGGDLVKPGASLTLTCTASGFSFSSSYWICWVRQAPGKGLEWIACI YAGSSGSTYY ASWAKGRFTISKTSSTTVTLQMTSLTAADTATYFCATAYAVSAPFGYTLFKYFELWGPGTL VTVSS

SEQ ID NO: 94 - 11889 rabbit VH (nucleic acid) cagtcgttggaggagtccgggggagacctggtcaagcctggggcatccctgacactcacct gcacagcctctggattctccttcagtagcagctactggatatgctgggtccgccaggctcc agggaaggggctggagtggatcgcatgcatttatgctggtagtagtggtagcacttactac gcgagctgggcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtgactctgc aaatgaccagtctgacagccgcggacacggccacctatttctgtgcgaccgcttatgctgt ttctgctccttttggttatactctttttaaatactttgaattgtggggcccaggcaccctg gtcaccgtctcgagc

SEQ ID NO: 95 - humanised 11889 VL

AW LTQSPSSLSASVGDRVTITCQASQNIYSDLAWYQQKPGKAPKLLIYDTSNLASGVPSR FSGSRSGTDFTLTISSLQPEDFATYYCQAYYYSSSSGDTTFGGGTKVEIK

SEQ ID NO: 96 - humanised 11889 VH

EVQLLESGGGLVQPGGSLRLSCAASGFSFSSSYWICWVRQAPGKGLEWIACI YAGSSGSTY YASWAKGRFTISKDSSKTTVYLQMNSLRAEDTAVYYCATAYAVSAPFGYTLFKYFELWGQG TLVTVSS

SEQ ID NO: 97 - humanised 11889 VH (CDRH1 and 2 variant) EVQLLESGGGLVQPGGSLRLSCAASGFSFSSSYWISWVRQAPGKGLEWIAS IYAGSSGSTY YASWAKGRFTISKDSSKTTVYLQMNSLRAEDTAVYYCATAYAVSAPFGYTLFKYFELWGQG TLVTVSS

SEQ ID NO: 98 - 11892 CDRL1

QASQSIGSNLA

SEQ ID NO: 99 - 11892 CDRL2

AASTLAS

SEQ ID NO: 100 - 11892 CDRL3

QCTYYANTYVAET

SEQ ID NO: 101 - 11892 CDRL3 variant

QQTYYANTYVAET

SEQ ID NO: 102 - 11892 CDRL3 variant

QSTYYANTYVAET

SEQ ID NO: 103 - 11892 CDRH1

GFSLSIYSMS

SEQ ID NO: 104 - 11892 CDRH2

IIYLGDRAYYASWAKG

SEQ ID NO: 105 - 11892 CDRH3

VAGYAGYGYAFYDAFDP

SEQ ID NO: 106 - 11892 CDRH3 variant

VAGYAGYGYAFYDAFEP

SEQ ID NO: 107 - 11892 CDRH3 variant

VAGYAGYGYAFYDAFNP

SEQ ID NO: 108 - 11892 rabbit VL (amino acid) DW MTQTPASVSEPVGGTVTIKCQASQSIGSNLAWYQQKPGQPPKLLIYAASTLASGVPSR

FKGSGSGTEFTLTINGVQCDDTATYYCQCTYYANTYVAETFGGGTEW VK

SEQ ID NO: 109 - 11892 rabbit VL (nucleic acid) gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagt caeca tcaagtgccaggccagtcagagcattggtagtaatttagcctggtatcagcagaaaccagg gcagcctcccaagctcctgatctatgctgcatccactctggcatctggggtcccatcgcgg ttcaaaggcagtggatctgggacagagttcactctcaccatcaacggcgtgcagtgtgacg atactgccacttactactgtcaatgtacttattatgctaatacttatgtggctgagacttt cggcggagggaccgaggtggtggtcaaa

SEQ ID NO: 110 - 11892 rabbit VH (amino acid)

QSMEESGGRLVTPGTPLTLTCTVSGFSLS IYSMSWVRQAPGKGLEWIGIIYLGDRAYYASW AKGRFTISKTSSTTVDLKITSPTTEDTATYFCARVAGYAGYGYAFYDAFDPWGPGTLVTVS S

SEQ ID NO: 111 - 11892 rabbit VH (nucleic acid) cagtcaatggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacct gcacagtctctggattctccctcagtatctattcaatgagctgggtccgccaggctccagg gaaggggctggaatggatcggaattatttatcttggtgatagggcatactacgcgagctgg gcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtggatctgaaaat caeca gtccgacaaccgaggacacggccacctatttctgtgccagagttgctggttatgctggtta tggttatgcgttctatgatgcttttgatccctggggcccaggcaccctggtcaccgtctcg age

SEQ ID NO: 112 - humanised 11892 VL

DVVMTQSPSSVSASVGDRVTITCQASQS IGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCQCTYYANTYVAETFGGGTKVEIK

SEQ ID NO: 113 - humanised 11892 VL (CDRL3 variant)

DVVMTQSPSSVSASVGDRVTITCQASQS IGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCQQTYYANTYVAETFGGGTKVEIK

SEQ ID NO: 114 - humanised 11892 VL (CDRL3 variant)

DVVMTQSPSSVSASVGDRVTITCQASQS IGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCQSTYYANTYVAETFGGGTKVEIK

SEQ ID NO: 115 - humanised 11892 VH

EVQLVESGGGLVQPGGSLRLSCAVSGFSLS IYSMSWVRQAPGKGLEWIGIIYLGDRAYYAS WAKGRFTISKDSSKTTVYLQMNSLRAEDTAVYYCARVAGYAGYGYAFYDAFDPWGQGTLVT VSS SEQ ID NO: 116 - humanised 11892 VH (CDRH3 variant)

EVQLVESGGGLVQPGGSLRLSCAVSGFSLS IYSMSWVRQAPGKGLEWIGIIYLGDRAYYAS WAKGRFTISKDSSKTTVYLQMNSLRAEDTAVYYCARVAGYAGYGYAFYDAFEPWGQGTLVT VSS

SEQ ID NO: 117 - humanised 11892 VH (CDRH3 variant)

EVQLVESGGGLVQPGGSLRLSCAVSGFSLS IYSMSWVRQAPGKGLEWIGIIYLGDRAYYAS WAKGRFTISKDSSKTTVYLQMNSLRAEDTAVYYCARVAGYAGYGYAFYDAFNPWGQGTLVT VSS

SEQ ID NO: 118 - 11881 CDRL1

QASQSIGSNLA

SEQ ID NO: 119 - 11881 CDRL2

AASTLAS

SEQ ID NO: 120 - 11881 CDRL3

QCTYYASTYVAET

SEQ ID NO: 121 - 11881 CDRL3 variant

QQTYYASTYVAET

SEQ ID NO: 122 - 11881 CDRL3 variant

QSTYYASTYVAET

SEQ ID NO: 123 - 11881 CDRH1

GFSLSIYSMS

SEQ ID NO: 124 - 11881 CDRH2

IIYLGDRAYYASWAKG

SEQ ID NO: 125 - 11881 CDRH3

VAGYAGYGYAFYDAFDP

SEQ ID NO: 126 - 11881 CDRH3 variant

VAGYAGYGYAFYDAFEP SEQ ID NO: 127 - 11881 CDRH3 variant

VAGYAGYGYAFYDAFNP

SEQ ID NO: 128 - 11881 rabbit VL (amino acid)

DW MTQTPASVSEPVGGTVTIKCQASQSIGSNLAWYQQKPGQPPKLLIYAASTLASGVPSR

FKGSGSGTEFTLTINGVQCDDTATYYCQCTYYASTYVAETFGGGTEW VK

SEQ ID NO: 129 - 11881 rabbit VL (nucleic acid) gatgttgtgatgacccagactccagcctccgtgtctgaacctgtgggaggcacagt caeca tcaagtgccaggccagtcagagcattggtagtaatttagcctggtatcagcagaaaccagg gcagcctcccaagctcctgatctatgctgcatccactctggcatctggggtcccatcgcgg ttcaaaggcagtggatctgggacagagttcactctcaccatcaacggcgtgcagtgtgacg atactgccacttactactgtcaatgtacttattatgctagtacttatgtggctgagacttt cggcggagggaccgaggtggtggtcaaa

SEQ ID NO: 130 - 11881 rabbit VH (amino acid)

SEQ ID NO: 131 - 11881 rabbit VH (nucleic acid) cagtcaatggaggagtccgggggtcgcctggtcacgcctgggacacccctgacactcacct gcacagtctctggattctccctcagtatctattcaatgagctgggtccgccaggctccagg gaaggggctggaatggatcggaattatttatcttggtgatagggcatactacgcgagctgg gcgaaaggccgattcaccatctccaaaacctcgtcgaccacggtggatctgaaaat caeca gtccgacaaccgaggacacggccacctatttctgtgccagagttgctggttatgctggtta tggttatgcgttctatgatgcttttgatccctggggcccaggcaccctggtcaccgtctcg age

SEQ ID NO: 132 - humanised 11881 VL

DVVMTQSPSSVSASVGDRVTITCQASQS IGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCQCTYYASTYVAETFGGGTKVEIK

SEQ ID NO: 133 - humanised 11881 VL (CDRL3 variant)

DVVMTQSPSSVSASVGDRVTITCQASQS IGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCQQTYYASTYVAETFGGGTKVEIK

SEQ ID NO: 134 - humanised 11881 VL (CDRL3 variant) DW MTQSPSSVSASVGDRVTITCQASQSIGSNLAWYQQKPGKAPKLLIYAASTLASGVPSR FSGSGSGTDFTLTISSLQPEDFATYYCQSTYYASTYVAETFGGGTKVEIK

SEQ ID NO: 135 - humanised 11881 VH

EVQLVESGGGLVQPGGSLRLSCAVSGFSLS IYSMSWVRQAPGKGLEWIGIIYLGDRAYYAS WAKGRFTISKDSSKTTVYLQMNSLRAEDTAVYYCARVAGYAGYGYAFYDAFDPWGQGTLVT VSS SEQ ID NO: 136 - humanised 11881 VH (CDRH3 variant)

SEQ ID NO: 137 - humanised 11881 VH (CDRH3 variant)

SEQ ID NO: 138 (66-3-9C HCDR1)

GFTFSSYNMN

SEQ ID NO: 139 (66-3-9C HCDR2)

SITTSSSYIYYAYSVKG SEQ ID NO: 140 (66-3-9C HCDR3)

FLGYSYGTNYYYYGMDV

SEQ ID NO: 141 (66-3-9C LCDR1)

RSSQSLLHSDGYNYLD

SEQ ID NO: 142 (66-3-9C LCDR2) LGSNRAS

SEQ ID NO: 143 (66-3-9C LCDR3)

MQALQTLT

SEQ ID NO: 144 (66-3-9C VH)

EVQLVESGGGLVKPGGSLRLSCAASG FTFSSYNMNWVRQAPGKGLEWVSSITTSSSYIYYA YSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARFLGYSYGTNYYYYGMDVWGQGTTV TVSS

SEQ ID NO: 145 (66-3-9C VL)

DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSDGYNYLDWYLQKPGQSPQLLI YLGSNRAS GVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTLTFGGGTKVEIK

SEQ ID NO: 146 (040 HCDR1)

GFTFSSYAMS

SEQ ID NO: 147 (040 HCDR2)

TISGSGGTTYYADSVKG

SEQ ID NO: 148 (040 HCDR3)

SYYYHSSGLLIRWDDMDV

SEQ ID NO: 149 (040 LCDR1)

RASQGIRNDLG

SEQ ID NO: 150 (040 LCDR2)

AASSLQS SEQ ID NO: 151 (040 LCDR3)

LQHSSYPWT

SEQ ID NO: 152 (040 VH)

QVQLVESGGGFVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSTISGSGGTTYYA

DSVKGRFTISRDNSKNTLYLEMITLRAEDTATYFCANSYYYHSSGLLIRWDDMDVWGQGTT

VTVSS

SEQ ID NO: 153 (040 VL)

DIQMTQSPSSLSASVGDRVTITCRASQGIRNDLGWYQQKPGKAPKRLIYAASSLQSGVPSR

FSGSGSGTEFTLTISSLQPEDFATYYCLQHSSYPWTFGQGTKVEIK

SEQ ID NO: 154 (6662 HCDR1)

GFGFSSAWMN

SEQ ID NO: 155 (6662 HCDR2)

RIKSKTDGGKTDYAAPVKG

SEQ ID NO: 156 (6662 HCDR3)

RIVLNGMDV

SEQ ID NO: 157 (6662 LCDR1)

TGSSSNIGAGYDVH

SEQ ID NO: 158 (6662 LCDR2)

ANNNRPS

SEQ ID NO: 159 (6662 LCDR3)

QSYDSRLSDGW

SEQ ID NO: 160 (6662 VH)

EVQLVESGGGLVKPGGSLRLSCAASGFGFSSAWMNWVRQAPGKGLEWVGRIKSKTDGGKTD

YAAPVKGRFIMSRDDSKNTLYLQMNSLKTEDAGVYYCTTRIVLNGMDVWGQGTLVTVSS SEQ ID NO: 161 (6662 VL)

QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYDVHWYQQLPGTAPKLLIYANNNRPSGVP DRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSRLSDGW FGGGTKLTVL

SEQ ID NO: 162 (66-3-9C heavy chain)

MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGSLRLSCAASGFTFSSYNMNWVRQAPG KGLEWVSSITTSSSYIYYAYSVKGRFTISRDNAKNSLYLQMNSLRAEDTAVYYCARFLGYS YGTNYYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVE PKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLD SDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

SEQ ID NO: 163 (66-3-9C kappa light chain) MGWSCIILFLVATATGVHSDIVMTQSPLSLPVTPGEPAS ISCRSSQSLLHSDGYNYLDWYL QKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQALQTLTFG GGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 164 (040 heavy chain)

MGWSCIILFLVATATGVHSQVQLVESGGGFVQPGGSLRLSCAASGFTFSSYAMSWVRQAPG KGLEWVSTISGSGGTTYYADSVKGRFT ISRDNSKNTLYLEMITLRAEDTATYFCANSYYYH SSGLLIRWDDMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPV TVSWNSGALTSGVHTFPAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKV EPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFN WYVDGVEVHNAKTKPREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEQ ID NO: 165 (040 kappa light chain)

MGWSCIILFLVATATGVHSDIQMTQSPSSLSASVGDRVTITCRASQGIRNDLGWYQQKPGK APKRLIYAASSLQSGVPSRFSGSGSGTEFTLTISSLQPEDFATYYCLQHSSYPWTFGQGTK VEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVT EQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

SEQ ID NO: 166 (6662 heavy chain)

MGWSCIILFLVATATGVHSEVQLVESGGGLVKPGGSLRLSCAASGFGFSSAWMNWVRQAPG KGLEWVGRIKSKTDGGKTDYAAPVKGRFIMSRDDSKNTLYLQMNSLKTEDAGVYYCTTRIV LNGMDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSW TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVW DVSHEDPEVKFNWYVDGVE VHNAKTKPREEQYNSTYRW SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPR EPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

SEQ ID NO: 167 (6662 lambda light chain)

MGWSCIILFLVATATGVHSQSVLTQPPSVSGAPGQRVT ISCTGSSSNIGAGYDVHWYQQLP GTAPKLLIYANNNRPSGVPDRFSGSKSGTSASLAITGLQAEDEADYYCQSYDSRLSDGW F GGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAG VETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

SEQ ID NO: 168 (66-3-9C variant LCDR1)

RSSQSLLHSEGYNYLD

SEQ ID NO: 169 (66-3-9C variant LCDR1)

RSSQSLLHSDSYNYLD SEQ ID NO: 170 (66-3-9C variant LCDR1)

RSSQSLLHSDAYNYLD

SEQ ID NO: 171 (6662 variant HCDR2)

RIKSKTEGGKTDYAAPVKG SEQ ID NO: 172 (6662 variant HCDR2)

RIKSKTDSGKTDYAAPVKG

SEQ ID NO: 173 (6662 variant HCDR2)

RIKSKTDAGKTDYAAPVKG

SEQ ID NO: 174 (6662 variant HCDR3) RIVLNSMDV

SEQ ID NO: 175 (6662 variant HCDR3)

RIVLNAMDV

SEQ ID NO: 176 (6662 variant LCDR3)

QSYDSRLSEGW SEQ ID NO: 177 (6662 variant LCDR3)

QSYDSRLSDSW SEQ ID NO: 178 (6662 variant LCDR3)

QSYDSRLSDAW

Claims

1. An anti-Ebola vims antibody that recognises an epitope on the Ebola vims glycoprotein comprising one or more residues selected from the group consisting of FI 32, P133, R134, C135, R136, Y137, V138, H139, K140, V141, S142 and G143 of SEQ ID

NO: 22.

2. The antibody of claim 1, which recognises an epitope comprising one or more residues selected from the group consisting of R134, C135 and R136 of SEQ ID NO: 22.

3. The antibody of claim 2, which recognises an epitope comprising R134 and R136 of SEQ ID NO: 22.

4. The antibody of claim 3, which recognises an epitope comprising all three of R134, C135 and R136.

5. The antibody of any one of the preceding claims, wherein the epitope further comprises:

(a) one of more residues selected from the group consisting of G102, E103, W104, A105, E106, N107 and C108 of SEQ ID NO: 22; and/or

(b) one of more residues selected from the group consisting of R605, W606, G607, G608, T609, C610 and H611 of SEQ ID NO: 22.

6. The antibody of any one of the preceding claims, which comprises one or more CDR sequences selected from the following:

(a) a CDRLl sequence of SEQ ID NO: 1;

(b) a CDRL2 sequence of SEQ ID NO: 2;

(c) a CDRL3 sequence of SEQ ID NO: 3, 4 or 5;

(d) a CDRH1 sequence of SEQ ID NO: 6;

(e) a CDRH2 sequence of SEQ ID NO: 7; and

(f) a CDRH3 sequence of SEQ ID NO: 8, 9 and 10.

7. The antibody of claim 6, which comprises a CDRH3 sequence of SEQ ID NO: 8, 9 or 10.

8. The antibody of claim 6 or 7 which comprises:

(a) a CDRL1 sequence of SEQ ID NO: 1;

(b) a CDRL2 sequence of SEQ ID NO: 2;

(c) a CDRL3 sequence selected from the group consisting of: SEQ ID NOs: 3, SEQ ID NO: 4 and SEQ ID NO: 5;

(d) a CDRH1 sequence of SEQ ID NO: 6;

(e) a CDRH2 sequence of SEQ ID NO: 7; and

(f) a CDRH3 sequence selected from the group consisting of: SEQ ID NO: 8, SEQ ID NO: 9 or SEQ ID NO: 10.

9. The antibody of any one of claims 6-8, which comprises a light chain variable region (VL) of SEQ ID NO: 11 and/or a heavy chain variable region (VH) of SEQ ID NO: 13.

10. The antibody of any one of claims 6-8, which comprises a light chain variable region with at least 90% identity to SEQ ID NO: 15, 16 or 17 and/or a heavy chain variable region with at least 90% identity to SEQ ID NO: 18, 19 or 20.

11. The antibody of claim 10, which comprises a light chain variable region selected from the group consisting of: SEQ ID NO: 15, SEQ ID NO: 16 and SEQ ID NO: 17, and/or a heavy chain variable region selected from the group consisting of: SEQ ID NO: 18, SEQ ID NO: 19 and SEQ ID NO: 20.

12. An antibody that binds to the same epitope as, or competes for binding with, an antibody as defined in any one of claims 6-11.

13. The antibody of any one of the preceding claims, which comprises an IgGl, IgG2, IgG3 or IgG4 constant region, optionally a human IgGl, IgG2, IgG3 or IgG4 constant region.

14. The antibody of any one of the preceding claims, which is cross-reactive for Ebola species Zaire, Sudan, Bundibugyo and Tai Forest.

15. The antibody of any one of the preceding claims, which has IC50 values of less than 2 pg/ml for each of Ebola species Zaire, Sudan and Bundibugyo and optionally IC90 values of less than 10 pg/ml for each of Ebola species Zaire, Sudan and Bundibugyo as determined in an assay measuring infection of cells using virus coated with the Ebola virus glycoprotein.

16. A nucleic acid or a pair of nucleic acids, optionally mRNA, encoding the heavy and light chains of an antibody as defined in any one of claims 1-15.

17. An expression vector comprising the nucleic acid(s) of claim 16 or a pair of expression vectors comprising the pair of nucleic acids of claim 15.

18. A host cell comprising the expression vector(s) of claim 17.

19. A method of producing an antibody of any one of claims 1-15, said method comprising culturing the host cell of claim 18 under conditions permitting production of the antibody and recovering the antibody so produced.

20. A pharmaceutical composition comprising an antibody of any one of claims 1-15 or a nucleic aci d/pair of nucleic acids of claim 16.

21. The pharmaceutical composition of claim 20, further comprising one or more additional anti-Ebola virus antibodies, or nucleic acids encoding one or more additional anti-Ebola virus antibodies.

22. The pharmaceutical composition of claim 21, wherein the anti -Ebola virus antibodies bind to different epitopes on the Ebola virus glycoprotein.

23. A method of treating, preventing or ameliorating Ebola virus infection, the method comprising administering a composition of claim 20, 21 or 22 to a subject in need thereof.

24. A composition of claim 20, 21 or 22 for use in a method of treating, preventing or ameliorating Ebola virus infection, the method comprising administering the composition to a subject in need thereof.

25. Else of a composition of claim 20, 21 or 22 in the manufacture of a medicament for treating, preventing or ameliorating Ebola virus infection.