[0295] Deparaffiniztion and rehydration: deparaffinized FFPE tissue by immersing in fresh xylene twice for 10 min each; rehydrated with a series of grade EtOH washes (100%, 95%, 85%, 75%) for 5 min, respectively.

[0296] Washed sections in dHaO for 3 min, and in PBS twice for 3 min each.

[0297] Epitope retrieval: heated the slides in the pH9.0 EDTA antigen retrieval solution (1 X) in a pressure cooker until boiling is initiated, followed with 3 min at high-temperature and high-pressure epitope retrieval. Cooled down to room temperature. Washed sections with PBS twice for 3 min.

[0298] Blocking: removed wash buffer, drew a hydrophobic circle around the tissue with a PAP pen. Blocked sections with peroxidase-blocking reagent for 15 min at room temperature. Washed sections with PBS twice for 3 min each.

[0299] Primary antibody incubation: conducted product-specific protocol for antibody dilution. Removed wash buffer and cover sections with 100-400pL primary antibody diluted in recommended antibody diluent. Incubated for 1 hour at 37°C in a humidified chamber. Washed sections in PBS twice for 3 min each.

[0300] Secondary antibody incubation: removed wash buffer and covered sections with 1 -3 drops of HRP conjugated goat anti-rabbit & mouse reagent. Incubated for 30 min at 37°C in a humidified chamber. Wash sections in PBS twice for 3 min each.

[0301] DAB staining: DAB-containing substrate working solution was prepared by mixing thoroughly 1 drop concentrated DAB solution per 1 mL DAB substrate buffer. Removed wash buffer and covered tissue section in 1 X DAB working solution for 3 min at room temperature. Washed sections in PBS twice for 3 min each.

[0302] Counterstain: completely submersed sections in hematoxylin and incubated for 1 min. Immersed slides in bluing solution. Rinsed slides in tap water for 3 min.

[0303] Dehydration and clearing: dehydrated with a series of grade EtOH washes (70%, 85%, 95%, 100%) for 3 min, respectively. Immersed in fresh xylene for twice for 10 min each.

[0304] Mounting: mounted sections with mounting medium.

2. FFPE Blocks Preparation on Cell line

[0305] Harvested cells respectively during the logarithmic growth period and counted cell number using Count-star.

[0306] About 5x10⁶ cells (1x10⁷) were collected after centrifuged at 1000 rpm at 4°C for 24 hour.

[0307] Discarded the supernatant, then added 30 mL 10% NBF into a 50mL tube with cell pellet.

[0308] Prepared cell suspension by reversing the tube several times. Then the tube was placed on bench flatwise and fixed the cells in 10%NBF for 30 min.

[0309] Centrifuged samples at 1000 rpm for 5 min at 4°C, discarded all the fixative carefully without disturbing cell pellet at the tube bottom, and placed samples in the incubator at 50°C for preheating.

[0310] Heated HistoGel in boiling water bath for 3-10 minutes and placed liquid HitoGel in the incubator at 50°C.

[0311] Added one or two drop of HistoGel on deposits of cell, vortexed for seconds to mix thoroughly. Then placed mixture on ice for 5-10 minutes to solidify the gel.

[0312] Placed the cell gel into a long hole embedding box. In a 250 mL beaker, rinsed the cell gel slowly with running water for 1 h, then added 50%, 75%, 85%, 95% ethanol to the cell block sequentially, let the cell gel soak for 1 h in each ethanol solution. Finally, soaked the cell gel in 100% ethanol twice, for 1 h each time.

[0313] Removed the cell blocks from 100% ethanol and soaked them in xylene twice, 1 h each time.

[0314] Soaked the cell block in liquid paraffin twice, 1 h each time, but can stay overnight.

[0315] Prepared FFPE block according to standard embedding process. Tissues were embedded in paraffin on Paraffin Embedding Station.

Example 1 - Hybridoma campaign for top clones

[0316] The PTK7 ECD protein was prepared in house and used as antigen. Monoclonal antibodies against PTK7 were developed by sequentially immunizing animals with PTK7-his antigen proteins with an adjuvant, and the experimental animals are Balb/c mice.

[0317] The animals were immunized with 100 pg of antigen per animal for the first shot, subsequently 50 pg of antigen per animal was used for immunization for the second and third shot, and 25 pg for the booster immunization. The immune adjuvant used in the experiments was Freund's Adjuvant (complete and incomplete). All animals were immunized by intraperitoneal injection. The mice with good immunized titer and serum immunohistochemistry (IHC) positive were chosen for booster immunization. After booster immunization, mice were sacrificed and soaked in 75% alcohol. The spleen was dissected out, grounded with a grinding rod, and filtered through a cell strainer to prepare a single cell suspension. The spleen cell suspension was centrifuged at 1 ,500 rpm for 5 min, and the supernatant was discarded. 5 mired blood cell lysate was added to lyse red blood cells at room temperature for 5 min and PBS was added to reach 20 ml_. After centrifugation at 1 ,500 rpm for 5 min, the supernatant was discarded. Viable cells were counted after resuspension. The Sp2/0 cells in the culture flask were collected and after centrifuged at 1 ,500 rpm for 5 min, the supernatant was discarded. Viable cells were counted after resuspension. The spleen cells were mixed with Sp2/0 cells at a ratio of 4:1 and subjected to centrifugation at 1 ,500 rpm for 5 min, the supernatant was discarded. The cells were resuspended in 20 mL electroporation buffer. After centrifugation at 1 ,500 rpm for 7 min, the supernatant was discarded, and the step was repeated once. The cells were resuspended with an appropriate amount of electroporation buffer to ensure the cell concentration of about 2x10⁷ cells/mL. The cell suspension was added to a 9 mL electroporation tank for fusion. After fusion, the cell suspension was transferred to 20 mL RPMI 1640 complete medium containing 20% FBS and then left at room temperature for 20 min. The fused cells were resuspended with RPMI 1640 medium containing 1 xHAT, 1xBIOMYC3, and 20% FBS. The cell suspension was added to several 96-well cell culture plates at 100 pL/well to ensure that the cell volume per well was about 5x10⁴ cells/well, and the plates was placed in a 37°C cell incubator. After 7 days, additional 100 pL of RPMI 1640 complete medium containing 20 % FBS, 1 xHAT, and 1 xBIOMYC-3 was added to each well. After 10 days of fusion, the cell culture supernatants from hybridoma parent clones were collected and used for screening by binding to human PTK7-his protein by ELISA and IHC method. The ELSIA and IHC both positive clones were subsequently chosen for further validation.

Example 2 - IHC method- mice serum validation

[0318] FFPE (Formalin-Fixed Paraffin-Embedded) sections of ovarian cancer were baked in a dry oven for 1 hour at 60°C, and then deparaffinized in fresh xylene for 10min and rehydrated through graded concentrations of ethanol (100%, 95%, 85%, 75%) to distilled water, the sections were washed with PBS twice for 3 min each. Epitope retrieval was performed by heating the sections in the pH 9.0 EDTA antigen retrieval solution (1 X) in a pressure cooker until boiling was initiated. The sections were cooled down to room temperature. The sections were washed with distilled water twice for 3 min. Endogenous peroxidase activity was blocked by incubating the sections in peroxidase-blocking reagent for 15 min at room temperature, then washed with PBS twice for 3 min each. Immunodetection of PTK7 was performed using the primary antibody (mice serum ) against PTK7 at dilution of 1 :500, 1 :2000, respectively. The sections were covered with 100-400 pL of primary antibody dilutions and incubated for 1 hour at 37°C. The sections were washed in PBS twice for 3 min each. Sections were then incubated with the HRP conjugated goat anti-rabbit &mouse reagent for 30 mins at 37°C and were visualized using DAB-containing substrate working solution as chromogen for 3 min at room temperature, resulting in brown staining. The sections were washed with PBS twice for 3 min each. Finally, the sections were counterstained (with hematoxylin and incubated for 1 min, rinsed in tap water for 3 min), dehydrated (through graded concentrations of ethanol (70%, 85%, 95%, 100%) for 3 min, respectively) and mounted in mounting medium.

Example 3 - IHC method-hybridoma screening (manual)

[0319] The sections of formalin-fixed, paraffine-embedded tumor tissue were baked in a dry oven for 1 hour at 60°C, and then deparaffinized in fresh xylene for 10 min and rehydrated through graded concentrations of ethanol (100%, 95%, 85%, 75%) to distilled water, the sections were washed with PBS twice for 3 mins. Epitope retrieval was performed by heating the slides in the pH9.0 EDTA antigen retrieval solution (1 X) in a pressure cooker until boiling was initiated. The sections cooled down to room temperature. The sections were washed with distilled water twice for 3 min. Endogenous peroxidase activity was blocked by incubating the sections in peroxidase- blocking reagent for 15 min at room temperature. Then the sections were washed with PBS twice for 3 min each. Immunodetection of PTK7 was performed using the primary antibody (hybridoma supernatant) against PTK7 and the sections were covered with 100-400 pL primary antibody dilutions and incubated for 1 hour at 37°C. The sections were washed in PBS twice for 3 min each. The sections were then incubated with the HRP conjugated goat anti-rabbit &mouse reagent for 30 mins at 37°C and were visualized using DAB-containing substrate working solution as chromogen for 3 min at room temperature, resulting in brown staining. The sections were washed with PBS twice for 3 min each. Finally, the sections were counterstained (with hematoxylin and incubated for 1 min, rinsed in tap water for 3 min), dehydrated (through graded concentrations of ethanol (70%, 85%, 95%, 100%) for 3 min, respectively) and mounted in mounting medium. The results are shown in FIGs. 1 A-1 D.

Example 4 - IHC method-subclone screening (manual)

[0320] The slices of formalin-fixed, paraffine-embedded tumor tissue were baked in a dry oven for 1 hour at 60°C, and then deparaffinized in fresh xylene for 10 min and rehydrated through graded concentrations of ethanol (100%, 95%, 85%, 75%) to distilled water. The sections were washed with PBS twice for 3 mins. Epitope retrieval was performed by heating the slides in the pH9.0 EDTA antigen retrieval solution (1 X) in a pressure cooker until boiling is initiated The sections cooled down to room temperature. The sections were washed with distilled water twice for 3 min. Endogenous peroxidase activity was blocked by incubating the sections in peroxidase- blocking reagent for 15 min at room temperature, then the sections were washed with PBS twice for 3 min each. Immunodetection of PTK7 was performed using the primary antibody (hybridoma supernatant) against PTK7 and the sections were covered with 100-400 pL primary antibody dilutions, and incubated for 1 hour at 37°C. The sections were washed in PBS twice for 3 min each. The sections were then incubated with the HRP conjugated goat anti-rabbit &mouse reagent for 30 mins at 37°C and were visualized using DAB-containing substrate working solution as chromogen for 3 min at room temperature, resulting in brown staining. The sections were washed with PBS twice for 3 min each. Finally, the sections were counterstained (with hematoxylin and incubated for 1 min, rinsed in tap water for 3 min), dehydrated (through graded concentrations of ethanol (70%, 85%, 95%, 100%) for 3 min, respectively) and mounted in mounting medium. Table 1 shows IHC screening results of top 8 subclones (underlined) in ovarian and breast cancers.

Table 1 : Top8 subclone IHC screening results in ovarian cancer and breast cancer

[0321 ] Selection of 206-M6-9H5 (M10091 -01 )(see FIGs. 2A-2B), 222-M6-2C6 (M10091 -04) (see FIGs. 3A-3B), 235-M6-14H10 (M10091 -05) (see FIGs. 4A-4B), 312-M10-5F7 (M10091 -08) (see FIGs. 5A-5B), 58-M1 -11 E5 (M10091 -06) (see FIGs. 6A-6B), 134-M6-17F7 (M10091-03) (see FIGs. 7A-7B), 236-M6-8G5 (M10091-07) (see FIGs. 8A-8B), 259-M6-11 E2 (M10091 -02) (see FIGs. 9A-9B), based on good and distinctive staining results.

[0322] Top 3 subclone IHC results: Table 2 shows the recommended concentration of top3 subclones for IHC test; table 3 shows the IDs and tissue sources of the tumor sections used in the titration assays of the top 3 subclones; table 4 shows the tissue types used in tissue microarray (TMA) of the top 3 subclones.

[0323] 11 E2: Ovarian cancer cell membrane positive, breast cancer cell membrane positive, tumor stroma positive, (see FIG.10)

[0324] 8G5: Ovarian cancer cell membrane positive, breast cancer cell membrane positive, tumor stroma positive, (see FIG.11 )

[0325] 5F7 : Ovarian cancer cell membranes are positive, breast cancer tumor cells are negative, and tumor stroma is positive, (see FIG.12)

[0326] CST : Ovarian cancer cell membranes are positive, breast cancer cell membranes are positive, and tumor stroma is positive, (see FIG.13)

Table 2: Top 3 subclone recommended IHC concentration

Table 3: Samples

Table 4: Tissue type

Example 5 - IHC method-Top clone optimization for TMA (manual)

[0327] The sections of formalin-fixed, paraffine-embedded tumor tissue were baked in a dry oven for 1 hour at 60°C, and then deparaffinized in fresh xylene for 10min and rehydrated through graded concentrations of ethanol (100%, 95%, 85%, 75%) to distilled water. The sections were washed with PBS twice for 3 mins. Epitope retrieval was performed by heating the sections in the pH9.0 EDTA antigen retrieval solution (1 X) in a pressure cooker until boiling was initiated. The sectioned cooled down to room temperature and then were washed with distilled water twice for 3 min. Endogenous peroxidase activity was blocked by incubating the sections in peroxidase- blocking reagent for 15 min at room temperature, the sections were washed with PBS twice for 3 min each. Immunodetection of PTK7 was performed using the primary antibody against PTK7 at recommend concentration (1 , 2, 4, 8 pg/mL, respectively ) and the sections were covered with 100-400 pL primary antibody dilutions and incubated for 1 hour at 37°C. The sections were washed in PBS twice for 3 min each. The sections were then incubated with the HRP conjugated goat anti-rabbit &mouse reagent for 30 mins at 37°C and were visualized using DAB-containing substrate working solution as chromogen for 3 min at room temperature, resulting in brown staining. The sections were washed with PBS twice for 3 min each. Finally, the sections were counterstained (with hematoxylin and incubated for 1 min, rinsed in tap water for 3 min), dehydrated (through graded concentrations of ethanol (70%, 85%, 95%, 100%) for 3 min, respectively) and mounted in mounting medium.

[0328] Subclone 11 E2 results are shown in FIGs. 14A-14D. Subclone 8G5 results are shown in FIGs. 15A-15D. Subclone 5F7 results are shown in FIGs. 16A-16D.

Example 6 - IHC method-TMA

[0329] One-millimeter cores from paraffin blocks of tumors were used to generate TMAs. Before staining, microarrays were baked in a dry oven for 1 hour at 60°C, these sections were deparaffinized in fresh xylene for 10min and rehydrated through graded concentrations of ethanol (100%, 95%, 85%, 75%) to distilled water, then washed with PBS twice for 3 min. Epitope retrieval was performed by heating the sections in the pH9.0 EDTA antigen retrieval solution (1 X) in a pressure cooker until boiling is initiated. The sections cooled down to room temperature then were washed with distilled water twice for 3 min. Endogenous peroxidase activity was blocked by incubating the sections in peroxidase-blocking reagent for 15 min at room temperature, then sections were washed with PBS twice for 3 min each. Immunodetection of PTK7 was performed using the primary antibody against PTK7 at recommend concentration (1 , 2, 2 pg/mL, respectively ) and the sections were covered with 100-400 pL of primary antibody dilutions and incubated for 1 hour at 37°C. The sections were washed in PBS twice for 3 min each. The sections were then incubated with the HRP conjugated goat anti-rabbit &mouse reagent for 30 mins at 37°C and were visualized using DAB-containing substrate working solution as chromogen for 3 min at room temperature, resulting in brown staining. The sections were washed with PBS twice for 3 min each. Finally, the sections were counterstained (with hematoxylin and incubate for 1 min, rinsed in tap water for 3 min), dehydrated (through graded concentrations of ethanol (70%, 85%, 95%, 100%) for 3 min, respectively) and mounted in mounting medium. Table 5 shows the TMA-Leica. Table 6 shows the TMA- manual.

Table 5: TMA - IHC by Leica Autostainer

Table 6: TMA - IHC by manual staining

[0330] Subclone 11 E2 TMA results are shown in FIGs. 17A-17B. Subclone 8G5 TMA results are shown in FIGs. 18A-18B. Subclone 5F7 TMA results are shown in FIGs.19A-19B.

Example 7 - IHC method- tumor cell blocks (manual)

[0331 ] The sections of formalin-fixed, paraffine-embedded tumor cells were baked in a dry oven for 1 hour at 60°C, then deparaffinized in fresh xylene for 10min and rehydrated through graded concentrations of ethanol (100%, 95%, 85%, 75%) to distilled water. The sections were washed with PBS twice for 3 min. Epitope retrieval was performed by heating the slides in the pH9.0 EDTA antigen retrieval solution (1 X) in a pressure cooker until boiling was initiated. The sections cooled down to room temperature The sections were washed with distilled water twice for 3 min. Endogenous peroxidase activity was blocked by incubating the sections in peroxidase-blocking reagent for 15 min at room temperature, then the sections washed with PBS twice for 3 min each. Immunodetection of PTK7 was performed using the primary antibody against PTK7 at recommend concentration (1 , 2, 2 pg/mL, respectively ) and the sections were covered with 100-400 pL primary antibody dilutions, incubation for 1 hour at 37°C. The sections were washed in PBS twice for 3 min each. The sections were then incubated with the HRP conjugated goat anti-rabbit &mouse reagent for 30 mins at 37°C and were visualized using DAB-containing substrate working solution as chromogen for 3 min at room temperature, resulting in brown staining. The sections were washed with PBS twice for 3 min each. Finally, the sections were counterstained (with hematoxylin and incubated for 1 min, rinsed in tap water for 3 min), dehydrated (through graded concentrations of ethanol (70%, 85%, 95%, 100%) for 3 min, respectively) and mounted in mounting medium.

[0332] Top 3 cell line IHC.

[0333] PTK7 antigen expression level: PA-1 >MDA-MB-453>MCF-7. Raji is negative control. Subclone 11 E2 results are shown in FIG. 20. Subclone 8G5 results are shown in FIG. 21. Subclone 5F7 results are shown in FIG. 22. PA-1 is a cancer cell line originally isolated from an ovarian cancer patient. MDA-MB-453 is a cancer cell line originally from a patient with metastatic breast carcinoma. MCF-7 is a cancer cell line isolated originally from a breast adenocarcinoma patient.

[0334] Having thus described the basic concepts, it may be rather apparent to those skilled in the art after reading this detailed disclosure that the foregoing detailed disclosure is intended to be presented by way of example only and is not limiting. Various alterations, improvements, and modifications may occur and are intended to those skilled in the art, though not expressly stated herein. These alterations, improvements, and modifications are intended to be suggested by this disclosure and are within the spirit and scope of the exemplary embodiments of this disclosure. [0335] Moreover, certain terminology has been used to describe embodiments of the present disclosure. For example, the terms “one embodiment,” “an embodiment,” and “some embodiments” mean that a particular feature, structure or characteristic described in connection with the embodiment is included in at least one embodiment of the present disclosure. Therefore, it is emphasized and should be appreciated that two or more references to “an embodiment” or “one embodiment” or “an alternative embodiment” in various portions of this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures or characteristics may be combined as suitable in one or more embodiments of the present disclosure.

[0336] Further, it will be appreciated by one skilled in the art, aspects of the present disclosure may be illustrated and described herein in any of a number of patentable classes or context including any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof.

[0337] Furthermore, the recited order of processing elements or sequences, or the use of numbers, letters, or other designations, therefore, is not intended to limit the claimed processes and methods to any order except as may be specified in the claims. Although the above disclosure discusses through various examples what is currently considered to be a variety of useful embodiments of the disclosure, it is to be understood that such detail is solely for that purpose and that the appended claims are not limited to the disclosed embodiments, but, on the contrary, are intended to cover modifications and equivalent arrangements that are within the spirit and scope of the disclosed embodiments.

[0338] Similarly, it should be appreciated that in the foregoing description of embodiments of the present disclosure, various features are sometimes grouped together in a single embodiment, figure, or description thereof to streamline the disclosure aiding in the understanding of one or more of the various embodiments. This method of disclosure, however, is not to be interpreted as reflecting an intention that the claimed subject matter requires more features than are expressly recited in each claim. Rather, claim subject matter lie in less than all features of a single foregoing disclosed embodiment.

FURTHER EMBODIMENTS - BINDING AGENTS

1 . A binding agent of protein tyrosine kinase 7 (PTK7), comprising: a heavy chain variable (VH) region and a light chain variable (VL) region, the VH region comprising complementarity determining regions HCDR1 , HCDR2 and HCDR3 disposed in heavy chain variable region framework regions and the VL region comprising LCDR1 , LCDR2 and LCDR3 disposed in light chain variable region framework regions, the VH and VL CDRs having amino acids sequences selected from the sets of amino acid sequences set forth in the group consisting of: a. SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, LVS, and SEQ ID NO: 7, respectively; b. SEQ ID NO: 10, SEQ ID NO: 11 , SEQ ID NO: 12, SEQ ID NO: 13, QMS, and SEQ ID NO: 14, respectively; and c. SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, RVS, and SEQ ID NO: 21 , respectively.

2. The binding agent of 1 , wherein the VH and VL regions have amino acid sequences that are selected from the pairs of amino acid sequences set forth in the group consisting of: a. SEQ ID NO: 1 and SEQ ID NO: 2, respectively; b. SEQ ID NO: 8 and SEQ ID NO:9, respectively; and c. SEQ ID NO: 15 and SEQ ID NO: 16, respectively.

3. The binding agent of 1 , wherein the VH and VL regions have amino acid sequences that are selected from the pairs of amino acid sequences set forth in the group consisting of: a. SEQ ID NO: 1 and SEQ ID NO: 2, respectively; b. SEQ ID NO: 8 and SEQ ID NO: 9, respectively; and c. SEQ ID NO: 15 and SEQ ID NO: 16, respectively, wherein the heavy and light chain framework regions are optionally modified with from 1 to 8 amino acid substitutions, deletions or insertions in the framework regions.

4. The binding agent of any one of claims 1 -3, wherein the framework regions are human framework regions.

5. The binding agent of any one of 1 -3, wherein the framework regions are mouse framework regions.

6. The binding agent of any one of 1 -5, wherein the binding agent is an antibody.

7. The binding agent of any one of 1-6, wherein one of 1 to 6, wherein the binding agent further comprises a heavy chain constant region.

8. The binding agent of any one of claims 1-6, wherein the binding agent is a monoclonal antibody. 9. The binding agent of 8, wherein the heavy chain constant region is of the IgG isotype.

10. The binding agent of 8, wherein the heavy chain constant region is a human lgG1 constant region or a human lgG4 constant region.

11 . The binding agent of 8, wherein the heavy chain constant region is a mouse lgG1 constant region, a mouse lgG2a constant region, a mouse lgG2c constant region, or a mouse lgG3 constant region.

12. The binding agent of 8, wherein the heavy chain constant region has the amino acid sequence set forth in SEQ ID NO: 22, 24, or 26.

13. The binding agent of any one of 1 to 11 , wherein the binding agent further comprises a light chain constant region.

14. The binding agent of 13, wherein the light chain constant region has the amino acid sequence set forth in SEQ ID NO: 23, 25, or 27.

15. The binding agent of 1 , wherein the binding agent is a monoclonal antibody comprising a VH region having the amino acid sequence set forth in SEQ ID NO: 1 , a VL region having the amino acid sequence set forth in SEQ ID NO: 2, a heavy chain constant region having the amino acid sequence set forth in SEQ ID NO: 22 and a light chain constant region having the amino acid sequence set forth in SEQ ID NO: 23.

16. The binding agent of 1 , wherein the binding agent is a monoclonal antibody comprising a VH region having the amino acid sequence set forth in SEQ ID NO: 8, a VL region having the amino acid sequence set forth in SEQ ID NO: 9, a heavy chain constant region having the amino acid sequence set forth in SEQ ID NO: 24 and a light chain constant region having the amino acid sequence set forth in SEQ ID NO: 25.

17. The binding agent of 1 , wherein the binding agent is a monoclonal antibody comprising a VH region having the amino acid sequence set forth in SEQ ID NO: 15, a VL region having the amino acid sequence set forth in SEQ ID NO: 16, a heavy chain constant region having the amino acid sequence set forth in SEQ ID NO: 26 and a light chain constant region having the amino acid sequence set forth in SEQ ID NO: 27.

FURTHER EMBODIMENTS - IHC METHODS:

A1 . A method of detecting a PTK7 expressing cell in a sample, comprising: contacting the sample with a detecting agent comprising the binding agent of any one of further numbered embodiments 1 -17, and detecting formation of a binding complex between the detecting agent and PTK7, wherein the detection of the binding complex is indicative of the presence of a PTK7 expressing cell in the sample. A2. The method of A1 , wherein the detecting agent further comprises a detectable marker that labels the binding agent.

A2.1 . The method of A2, wherein the detectable marker includes biotin or an enzyme marker.

A2.2. The method of A2.1 , wherein the enzyme marker includes horseradish peroxidase (HRP) or alkaline phosphatase (AP).

A2.3. The method of any one of A2 - A2.2, wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: adding a partner agent of the detectable marker; and detecting a reaction of the detectable marker with the partner agent as an indication of the formation of the binding complex.

A2.4. The method of A2, wherein the detectable marker includes a fluorescent tag.

A2.5. The method of A2.4, wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: detecting a signal from the fluorescent tag as an indication of the formation of the binding complex.

A3. The method of A1 , wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: contacting the sample with a secondary antibody that is labeled and configured to bind to the binding complex; and detecting a signal from the secondary antibody as an indication of the formation of the binding complex.

A3.1 . The method of A3, wherein the secondary antibody is labeled with a detectable marker.

A3.2. The method of A3.1 , wherein the detectable marker includes biotin or an enzyme marker.

A3.3. The method of any one of A3.1 -A3.2, wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: adding a partner agent of the detectable marker; and detecting a reaction of the detectable marker with the partner agent as an indication of the formation of the binding complex.

A3.4. The method of claim A3.1 , wherein the detectable marker includes a fluorescent tag.

A3.5. The method of claim A3.4, wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: detecting a signal from the fluorescent tag as an indication of the formation of the binding complex.

A3.6. The method of A3.4, wherein the fluorescent tag includes fluorescein (FITC), phycoerythrin (PE), preferable R-PE, or allophycocyanin (APC).

A4. The method of A1 , wherein the sample is from a tissue of a subject. A4.1 . The method of A4, wherein the tissue is selected from the group consisting of tonsil, appendix, breast, ovary, colon, prostate, skin, lung, uterus, cervix, kidney, pancreas, bladder, brain, thyroid, ear, nose, throat, and esophagus.

A4.2. The method of any one of A4-A4.1 , wherein the tissue is selected from the group consisting of breast, ovary, colon, prostate, skin, lung, uterus, esophagus, and bladder.

A4.3. The method of any one of A4-A4.2, wherein the tissue is breast.

A4.4. The method of any one of A4-A4.2, wherein the tissue is ovary.

A5. The method of A1 , wherein the sample is from a subject and the presence of PTK7 expressing cell indicates that the subject is suffering from a disease.

A5.1 . The method of A5.1 , wherein the sample is from a subject and the method further comprises: quantifying binding PTK expressing cells in the sample to obtain a quantification result, wherein the quantification result over a pre-determined threshold indicates that the subject is suffering from a disease.

A5.2. The method of A5 or A5.1 , wherein the disease is cancer.

A5.3. The method of A5 or A5.1 , wherein the disease is selected from the group consisting of breast cancer, ovarian cancer, colon cancer, prostate cancer, melanoma, lung cancer, esophageal cancer, gastric cancer, endometrial cancer, head and neck cancer, and bladder cancer.

A5.4. The method of claim A5 or A5.1 , wherein the disease is breast cancer.

A5.5. The method of claim A5 or A5.1 , wherein the disease is ovarian cancer.

A5.6. The method of claim A5 or A5.1 , wherein the disease is lung squamous cells carcinoma or lung adenocarcinoma.

FURTHER EMBODIMENTS - DISEASE DIAGNOSIS, TREATMENT AND MONITORING

B1 . A method of determining whether a subject is suffering from a disease, comprising: obtaining a sample from the subject; contacting the sample with a detecting agent comprising the binding agent of any one of further numbered embodiments 1 -17, and determining whether the subject is suffering from the disease by detecting and/or quantifying formation of binding complexes between the detecting agent and PTK7, wherein the presence of the binding complexes or a level of the binding complexes over a pre-determined threshold indicates that the subject is suffering from the disease.

B1 .1 . The method of B1 , wherein the disease is cancer.

B2. A method of treating a disease, comprising: obtaining a sample from a subject; contacting the sample with a detecting agent comprising the binding agent of any one of 1 -17; determining whether the subject is suffering from the disease by detecting and/or quantifying formation of binding complexes between the detecting agent and PTK7, wherein the presence of the binding complexes or a level of the binding complexes over a pre-determined threshold indicates that the subject is suffering from the disease; and treating the disease.

B2.1 . The method of B2, wherein the disease is cancer.

B2.2. The method of B2.1 , wherein the disease is selected from the group consisting of breast cancer, ovarian cancer, colon cancer, prostate cancer, melanoma, lung cancer, esophageal cancer, gastric cancer, endometrial cancer, head and neck cancer, and bladder cancer.

B2.3. The method of any one of B2-B2.2, wherein the treatment includes administering chemotherapy to the subject.

B2.4. The method of any one of B2-B2.2, wherein the treatment includes administering hormone therapy to the subject.

B2.5. The method of any one of B2-B2.2, wherein the treatment includes administering radiation therapy to the subject.

B2.6. The method of any one of cB2-B2.2, wherein the treatment includes administering immunotherapy to the subject.

B2.7. The method of any one of B2-B2.2, wherein the treatment includes administering stem cell therapy to the subject.

B2.8. The method of any one of B2-B2.2, wherein the treatment includes administering targeted therapy to the subject.

B2.9. The method of any one of B2-B2.2, wherein the treatment includes performing surgery on the subject.

B2.10. The method of any one of B2-B2.2, wherein the disease is breast cancer and the treatment includes lumpectomy or mastectomy.

B2.11. The method of any one of B2-B2.2, wherein the disease is ovarian cancer and the treatment includes hysterectomy or salpingo-oophorectomy.

B3. A method of monitoring progression of a disease in a subject, comprising the steps of:

(a) obtaining a sample from the subject;

(b) contacting the sample with a detecting agent comprising the binding agent of any one of further numbered embodiments 1-17;

(c) assessing formation of binding complexes between the detecting agent and PTK7 to obtaining a first assessment result;

(d) repeating steps (a), (b), and (c) using a sample from the subject at a subsequent point in time to obtaining a second assessment result; and (e) comparing the second assessment result to the first assessment result to determine the pression of the disease in the subject.

B3.1 . The method of B3, wherein the disease is cancer.

FURTHER EMBODIMENTS - KITS

C1 . A kit for detecting PTK7 expressing cells in a sample, the kit comprising a detecting agent that comprises the binding agent of any one of further numbered embodiments 1 -17.

C2. The kit of claim C1 , further comprising a second antibody that is labeled and configured to bind to a binding complex that is formed by the detecting agent and PTK7.

SEQUENCE LISTING

SEQ ID NO: 1

QVTLKESGPGILQPSQTLSLTCSFSGFSLNTFGMGVSWIRQPSGNGLEWLAHIYWDDGKDYNP

SLKSRLTISKDTSNNQVFLKITTVDTTDTATYYCAYGFAYWGQGTLVTVSA

SEQ ID NO: 2

DVVMTQTPLTLSVTIGQPASISCKSSQSLLSINGKTYLNWLLQRPGQSPKRLIHLVSKLDSGVPD

RFTGSGSGTDFTLKISRVEAEDLGVYYCVQGTHFPHTFGGGTKLEIK

SEQ ID NO: 3 GFSLNTFGMG

SEQ ID NO: 4 IYWDDGK

SEQ ID NO: 5 AYG FAY

SEQ ID NO: 6 QSLLSINGKTY

SEQ ID NO: 7 VQGTHFPHT

SEQ ID NO: 8

EVMLVESGGGSAKPGGSLKLSCEASGFTFSSYAMSWVRQTPGKRLEWVASISNGGSYTNYPD

SVKGRFTISRDNAKNTLYLQMTSLRSEDTAIYYCSNTGTSYYGFEYWGQGTTLTVSS

SEQ ID NO: 9

DIVMTQAAFSSPVTLGTSASISCRSSESLLYSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPD

RFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLELPYTFGGGTRLEIK

SEQ ID NO: 10 GFTFSSYA

SEQ ID NO: 11 ISNGGSYT

SEQ ID NO: 12 SNTGTSYYGFEY

SEQ ID NO: 13 ESLLYSNGITY

SEQ ID NO: 14 AQNLELPYT

SEQ ID NO: 15

EVQLQQSGPVLVKPGASVKISCKASGYTFTDYNMHWVRQSHGKSLEWIGYIYPYNGGTGYNQ KFKSKATLTVDNSSNTAYLELRSLTSEDSAVYYCARGGWYFDVWGAGTTVTVSS

SEQ ID NO: 16

DAVMTQTPLSLPVSLGDQASISCRSSQSLENSNGNTYLNWYLQKPGQSPQLLIYRVSNRFSGV

LDRFSGSGSGTDFTLKISRVEAEDLGVYFCLQITHVPFTFGSGTKLEIK

SEQ ID NO: 17 GYTFTDYN

SEQ ID NO: 18 IYPYNGGT

SEQ ID NO: 19 ARGGWYFDV

SEQ ID NO: 20 QSLENSNGNTY

SEQ ID NO: 21 LQITHVPFT

SEQ ID NO: 22

AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTL

SSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTI

TLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNG

KEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEW

QWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSH

SPGK

SEQ ID NO: 23

RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKD

STYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

SEQ ID NO: 24

AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTL

SSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTI

TLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNG

KEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEW

QWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSH

SPGK

SEQ ID NO: 25

RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKD

STYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

SEQ ID NO: 26

AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTL

SSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTI

TLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNG

KEFKCRVNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEW

QWNGQPAENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSH SPGK SEQ ID NO: 27

RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKD

STYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

SEQ ID NO: 28

QVTLKESGPGILQPSQTLSLTCSFSGFSLNTFGMGVSWIRQPSGNGLEWLAHIYWDDGKDYNP

SLKSRLTISKDTSNNQVFLKITTVDTTDTATYYCAYGFAYWGQGTLVTVSAAKTTPPSVYPLAPG

SAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSSTWPS

ETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDIS

KDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFP

APIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNT

QPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK

SEQ ID NO: 29

DVVMTQTPLTLSVTIGQPASISCKSSQSLLSINGKTYLNWLLQRPGQSPKRLIHLVSKLDSGVPD

RFTGSGSGTDFTLKISRVEAEDLGVYYCVQGTHFPHTFGGGTKLEIKRADAAPTVSIFPPSSEQL

TSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYER

HNSYTCEATHKTSTSPIVKSFNRNEC

SEQ ID NO: 30

EVMLVESGGGSAKPGGSLKLSCEASGFTFSSYAMSWVRQTPGKRLEWVASISNGGSYTNYPD

SVKGRFTISRDNAKNTLYLQMTSLRSEDTAIYYCSNTGTSYYGFEYWGQGTTLTVSSAKTTPPS

VYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVP

SSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVT

CVVVDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCR

VNSAAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQP

AENYKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK

SEQ ID NO: 31

DIVMTQAAFSSPVTLGTSASISCRSSESLLYSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPD

RFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLELPYTFGGGTRLEIKRADAAPTVSIFPPSSEQL

TSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYER

HNSYTCEATHKTSTSPIVKSFNRNEC

SEQ ID NO: 32

EVQLQQSGPVLVKPGASVKISCKASGYTFTDYNMHWVRQSHGKSLEWIGYIYPYNGGTGYNQ

KFKSKATLTVDNSSNTAYLELRSLTSEDSAVYYCARGGWYFDVWGAGTTVTVSSAKTTPPSVYP

LAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFPAVLQSDLYTLSSSVTVPSST

WPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVV

VDISKDDPEVQFSWFVDDVEVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNS

AAFPAPIEKTISKTKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAEN YKNTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK

SEQ ID NO: 33

DAVMTQTPLSLPVSLGDQASISCRSSQSLENSNGNTYLNWYLQKPGQSPQLLIYRVSNRFSGV

LDRFSGSGSGTDFTLKISRVEAEDLGVYFCLQITHVPFTFGSGTKLEIKRADAAPTVSIFPPSSEQ

LTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYE

RHNSYTCEATHKTSTSPIVKSFNRNEC

Claims

WHAT IS CLAIMED IS:

1 . A binding agent for protein tyrosine kinase 7 (PTK7), the binding agent comprising: a heavy chain variable (VH) region and a light chain variable (VL) region, the VH region comprising complementarity determining regions HCDR1 , HCDR2 and HCDR3 disposed in heavy chain variable region framework regions and the VL region comprising LCDR1 , LCDR2 and LCDR3 disposed in light chain variable region framework regions, wherein the VH and VL are selected from one of the following pairs of VH and VL set out in a. to c. below: a. the VH comprising a HCDR1 comprising SEQ ID NO: 3, a HCDR2 comprising SEQ ID NO: 4, and a HCDR3 comprising SEQ ID NO: 5, and the VL comprising a LCDR1 comprising SEQ ID NO: 6, a LCDR2 comprising LVS, and a LCDR3 comprising SEQ ID NO: 7; b. the VH comprising a HCDR1 comprising SEQ ID NO: 10, a HCDR2 comprising SEQ ID NO: 11 , and a HCDR3 comprising SEQ ID NO: 12, and the VL comprising a LCDR1 of SEQ ID NO: 13, a LCDR2 comprising QMS, and a LCDR3 comprising SEQ ID NO: 14; and c. the VH comprising a HCDR1 comprising SEQ ID NO: 17, a HCDR2 comprising SEQ ID NO: 18, and a HCDR3 comprising SEQ ID NO: 19, and the VL comprising a LCDR1 of SEQ ID NO: 20, a LCDR2 comprising RVS, and a LCDR3 comprising SEQ ID NO: 21 (LCDR3).

2. The binding agent of claim 1 , wherein the VH and VL region are selected from one of the following pairs of VH and VL regions: a. a VH comprising SEQ ID NO: 1 and a VL comprising SEQ ID NO: 2; b. a VH comprising SEQ ID NO: 8 and a VL comprising SEQ ID NO: 9; and c. a VH comprising SEQ ID NO: 15 and a VL comprising SEQ ID NO: 16.

3. The binding agent of claim 1 , wherein the VH and VL regions have amino acid sequences that one of the following pairs of VH and VL sequences set forth below: a. a VH comprising SEQ ID NO: 1 and a VL comprising SEQ ID NO: 2; b. a VH comprising SEQ ID NO: 8 and a VL comprising SEQ ID NO: 9; and c. a VH comprising SEQ ID NO: 15 and a VL comprising SEQ ID NO: 6, wherein the heavy and light chain framework regions are optionally modified with from 1 to 8 amino acid substitutions, deletions or insertions in the framework regions and the antibody retains the ability to bind PTK7.

5. The binding agent of any one of claims 1 -3, wherein the framework regions are mouse framework regions.

6. The binding agent of any one of claims 1 -5, wherein the binding agent is an antibody.

7. The binding agent of any one of claims 1 -6, wherein the binding agent is a monoclonal antibody.

8. The binding agent of any of the preceding claims, wherein the binding agent comprises a heavy chain comprising the VH and a heavy chain constant region.

9. The binding agent of claim 8, wherein the heavy chain constant region is of the IgG isotype.

10. The binding agent of claim 8, wherein the heavy chain constant region is a human lgG1 constant region or a human lgG4 constant region.

11 . The binding agent of claim 8, wherein the heavy chain constant region is a mouse lgG1 constant region, a mouse lgG2a constant region, a mouse lgG2c constant region, or a mouse lgG3 constant region.

12. The binding agent of claim 8, wherein the heavy chain constant region has the amino acid sequence set forth in SEQ ID NO: 22, 24, or 26.

13. The binding agent of any of the preceding claims, wherein the binding agent comprises a light chain comprising the VL and a light chain constant region.

14. The binding agent of claim 13, wherein the light chain constant region has the amino acid sequence set forth in SEQ ID NO: 23, 25, or 27.

15. The binding agent of claim 1 , wherein the binding agent is an antibody comprising: a. a heavy chain comprising a VH region having the amino acid sequence set forth in SEQ ID NO: 1 and a heavy chain constant region having the amino acid sequence set forth in SEQ ID NO: 22; and b. a light chain comprising a VL region having the amino acid sequence set forth in SEQ ID NO: 2 and a light chain constant region having the amino acid sequence set forth in SEQ ID NO: 23.

16. The binding agent of claim 1 , wherein the binding agent is an antibody comprising: a. a heavy chain comprising a VH region having the amino acid sequence set forth in SEQ ID NO: 8 and a heavy chain constant region having the amino acid sequence set forth in SEQ ID NO: 24; and b. a light chain comprising a VL region having the amino acid sequence set forth in SEQ ID NO: 9 and a light chain constant region having the amino acid sequence set forth in SEQ ID NO: 25.

17. The binding agent of claim 1 , wherein the binding agent is an antibody comprising: a. a heavy chain comprising a VH region having the amino acid sequence set forth in SEQ ID NO: 15 and a heavy chain constant region having the amino acid sequence set forth in SEQ ID NO: 26; and b. a light chain comprising a VL region having the amino acid sequence set forth in SEQ ID NO: 16 and a light chain constant region having the amino acid sequence set forth in SEQ ID NO: 27.

18. The binding agent of claim 1 comprising a heavy chain of SEQ ID NO: 28 and a light chain of SEQ ID NO: 29.

19. The binding agent of claim 1 comprising a heavy chain of SEQ ID NO: 30 and a light chain of SEQ ID NO: 31.

20. The binding agent of claim 1 comprising a heavy chain of SEQ ID NO: 32 and a light chain of SEQ ID NO: 33.

21 . A method of detecting a PTK7 expressing cell in a sample, comprising: contacting the sample with a detecting agent comprising the binding agent of any one of claims 1 -20, and detecting formation of a binding complex between the detecting agent and PTK7, wherein the detection of the binding complex is indicative of the presence of a PTK7 expressing cell in the sample.

22. The method of claim 21 , wherein the detecting agent further comprises a detectable marker that labels the binding agent.

23. The method of claim 21 , wherein the detectable marker includes biotin or an enzyme marker.

24. The method of claim 23, wherein the enzyme marker includes horseradish peroxidase (HRP) or alkaline phosphatase (AP).

25. The method of any one of claims 21 -24, wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: adding a partner agent of the detectable marker; and detecting a reaction of the detectable marker with the partner agent as an indication of the formation of the binding complex.

26. The method of claim 22, wherein the detectable marker includes a fluorescent tag.

27. The method of claim 21 , wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: detecting a signal from the fluorescent tag as an indication of the formation of the binding complex.

28. The method of claim 21 , wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: contacting the sample with a secondary antibody that is labeled and configured to bind to the binding complex; and detecting a signal from the secondary antibody as an indication of the formation of the binding complex.

29. The method of claim 28, wherein the secondary antibody is labeled with a detectable marker.

30. The method of claim 29, wherein the detectable marker includes biotin or an enzyme marker.

31 . The method of any one of claims 25 to 30, wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: adding a partner agent of the detectable marker; and detecting a reaction of the detectable marker with the partner agent as an indication of the formation of the binding complex.

32. The method of claim 25, wherein the detectable marker includes a fluorescent tag.

33. The method of claim 32, wherein the detecting the formation of the binding complex between the detecting agent and PTK7 comprises: detecting a signal from the fluorescent tag as an indication of the formation of the binding complex.

34. The method of claim 33 wherein the fluorescent tag includes fluorescein (FITC), phycoerythrin (PE), preferable R-PE, or allophycocyanin (APC).

35. The method of claim 21 , wherein the sample is from a tissue of a subject.

36. The method of claim 35, wherein the tissue is selected from the group consisting of tonsil, appendix, breast, ovary, colon, prostate, skin, lung, uterus, cervix, kidney, pancreas, bladder, brain, thyroid, ear, nose, throat, and esophagus.

37. The method of claim 35, wherein the tissue is selected from the group consisting of breast, ovary, colon, prostate, skin, lung, uterus, esophagus, and bladder.

38. The method of any one of claims 35 to 37, wherein the tissue is breast.

39. The method of any one of claims 35 to 37, wherein the tissue is ovary.

40. The method of claim 21 , wherein the sample is from a subject and the presence of PTK7 expressing cell indicates that the subject is suffering from a disease.

41 . The method of claim 37, wherein the sample is from a subject and the method further comprises: quantifying binding PTK expressing cells in the sample to obtain a quantification result, wherein the quantification result over a pre-determined threshold indicates that the subject is suffering from a disease.

42. The method of claim 40 or 41 , wherein the disease is cancer.

43. The method of claim 40 or 41 , wherein the disease is selected from the group consisting of breast cancer, ovarian cancer, colon cancer, prostate cancer, melanoma, lung cancer, esophageal cancer, gastric cancer, endometrial cancer, head and neck cancer, and bladder cancer.

44. The method of claim 42 or 43, wherein the disease is breast cancer.

45. The method of claim 42 or 43, wherein the disease is ovarian cancer.

46. The method of claim 42 or 43, wherein the disease is lung squamous cells carcinoma or lung adenocarcinoma.

47. The method of any one of claims 21 to 46, wherein the method is an immunohistochemistry method.

48. Use of a binding agent according to any one of claims 1 to 20 to detect PTK7 protein.

49. A method of determining whether a subject is suffering from a disease, comprising: obtaining a sample from the subject; contacting the sample with a detecting agent comprising the binding agent of any one of claims 1 -20, and determining whether the subject is suffering from the disease by detecting and/or quantifying formation of binding complexes between the detecting agent and PTK7, wherein the presence of the binding complexes or a level of the binding complexes over a pre-determined threshold indicates that the subject is suffering from the disease.

50. The method of claim 49 wherein the disease is cancer.

51 . A method of treating a disease in a subject, wherein the subject has been determined to have the disease by the method of claim 40.

52. The method of claim 51 , wherein the binding agent comprises VH and VL regions comprising amino acid sequences set forth in: SEQ ID NO: 1 and SEQ ID NO: 2, respectively; SEQ ID NO: 8 and SEQ ID NO:9, respectively; or SEQ ID NO: 15 and SEQ ID NO: 16, respectively.

53. A method of treating a disease, comprising: obtaining a sample from a subject; contacting the sample with a detecting agent comprising the binding agent of any one of claims 1 -20; determining whether the subject is suffering from the disease by detecting and/or quantifying formation of binding complexes between the detecting agent and PTK7, wherein the presence of the binding complexes or a level of the binding complexes over a pre-determined threshold indicates that the subject is suffering from the disease; and treating the disease.

54. The claims of 53, wherein the disease is cancer.

55. The method of claim 54, wherein the disease is selected from the group consisting of breast cancer, ovarian cancer, colon cancer, prostate cancer, melanoma, lung cancer, esophageal cancer, gastric cancer, endometrial cancer, head and neck cancer, and bladder cancer.

56. The method of any one of claims 53 to 55, wherein the treatment includes administering chemotherapy to the subject.

57. The method of any one of claims 53 to 56, wherein the treatment includes administering hormone therapy to the subject.

58. The method of any one of claims 53 to 56, wherein the treatment includes administering radiation therapy to the subject.

59. The method of any one of claims 53 to 56, wherein the treatment includes administering immunotherapy to the subject.

60. The method of any one of claims 53 to 56, wherein the treatment includes administering stem cell therapy to the subject.

61 . The method of any one of claims 53 to 56, wherein the treatment includes administering targeted therapy to the subject.

62. The method of any one of claims 53 to 56, wherein the treatment includes performing surgery on the subject.

63. The method of any one of claims 53 to 56, wherein the disease is breast cancer and the treatment includes lumpectomy or mastectomy.

64. The method of any one of claims 53 to 56, wherein the disease is ovarian cancer and the treatment includes hysterectomy or salpingo-oophorectomy.

65. A method of monitoring progression of a disease in a subject, comprising the steps of:

(a) contacting a sample from the subject with a detecting agent comprising the binding agent of any one of claims 1 -20;

(b) assessing formation of binding complexes between the detecting agent and PTK7 to obtaining a first assessment result;

(c) repeating steps (a), (b), and (c) using a sample from the subject at a subsequent point in time to obtaining a second assessment result; and

(d) comparing the second assessment result to the first assessment result to determine the pression of the disease in the subject.

66. The method of 65, wherein the disease is cancer.

67. A kit for detecting PTK7 expressing cells in a sample, the kit comprising a detecting agent that comprises the binding agent of any one of claims 1 -21 .

68. The kit of claim 67, further comprising a second antibody that is labeled and configured to bind to a binding complex that is formed by the detecting agent and PTK7.