Fibroblast targeting molecule

Abstract

Provided herein are antigen-binding proteins that target cancer-associated fibroblasts. Further provided are methods of treatment comprising administering a pharmaceutical composition comprising an antigen-binding protein as described herein.

Description

[0001] Cross-references to related applications This application claims the benefit of U.S. Provisional Patent Application No. 63 / 596,210, filed November 3, 2023, which is hereby incorporated by reference in its entirety.

[0002] By incorporating materials submitted electronically. By reference to the full text of this paper is a computer-readable nucleotide / amino acid sequence list submitted concurrently with this paper, identified as follows: a 466-byte XML file named “FIBROBLAST TARGETING MOLECULES Sequence Listing”; created on November 3, 2023. Technical Field

[0003] This invention relates to antigen-binding proteins that target cancer-associated fibroblasts (CAFs) for the treatment of cancer. Background Technology

[0004] Fibroblast activator protein α (FAP or FAPα) (also known as Seprase) is a type II integrated membrane serine peptidase. FAP belongs to the dipeptidyl peptidase IV family (Yu et al., FEBS J. [Journal of the Federation of European Biochemical Societies] 277, 1126-1144 (2010)). It is a 170 kDa homodimer containing two N-glycosylated subunits with a large C-terminal extracellular domain, in which the enzyme's catalytic domain is located (Scanlan et al., Proc Natl Acad Sci USA [Proceedings of the National Academy of Sciences of the United States of America] 91: 5657-5661 (1994); Wonganu et al., Biochim Biophys Acta [Chinese Journal of Biochemistry and Biophysics] 1858(8):1876-82 (2016)). Glycosylated FAP possesses both prolyl dipeptidyl peptidase and gelatinase activities (Sun et al., Protein Expression and Purification 24, 274-281 (2002)). Homologs of human FAP exist in several species, including mice and cynomolgus monkeys (Macaca fascicularis).

[0005] FAP is selectively expressed in reactive stromal fibroblasts in over 90% of the epithelial malignancies examined (primary and metastatic) (including lung, colorectal, bladder, ovarian, and breast cancer) and in malignant mesenchymal cells of bone and soft tissue sarcomas, whereas it is not normally present in normal adult tissues (Brennen et al., Mol. CancerTher. [Molecular Cancer Therapeutics] 11(2): 257–266 (2012); Garin-Chesa et al., Proc Natl Acad Sci USA [Proceedings of the National Academy of Sciences] 87, 7235-7239 (1990); Rettig et al., Cancer Res. [Cancer Research] 53:3327–3335 (1993); Rettig et al., Proc Natl Acad Sci USA [Proceedings of the National Academy of Sciences] 85, 3110-3114 (1988)). FAP is also expressed in some malignant tumor cells. Because it is expressed in many common cancers and its expression is limited in normal tissues, FAP is considered a promising antigen target for imaging, diagnosis and therapy of a variety of cancers.

[0006] Various approaches, including monoclonal antibodies targeting FAP and small-molecule inhibitors of FAP enzyme activity, have been devised to leverage the selective expression of FAP in the tumor stroma for clinical benefit. For example, siroizumab (a humanized antibody derived from an F19 mouse antibody) has been investigated for the treatment of metastatic colorectal cancer and non-small cell lung cancer (Scott et al., Clin. Cancer Res. 9, 1639–1647 (2003)). Several FAP-targeting molecules have been reported to inhibit FAP enzyme activity. For example, scFv E3 inhibits both FAP enzyme activity and biological function (Zhang et al., FASEB J. 2013 Feb; 27(2): 581–589). However, the role of FAP in tumor biology is complex and not fully understood, and therefore the potential effects of FAP inhibition in tumor biology are also not fully understood.

[0007] FAP expression in the tumor stroma has also spurred attempts to develop locally activated prodrugs. Brennen et al. (Mol. Cancer Ther. [Molecular Cancer Therapeutics] 11(2): 257–266 (2012)) discussed a method to develop FAP-activated prodrugs to target and activate cytotoxic compounds within the tumor stroma by utilizing the restricted expression and unique substrate preference of FAP.

[0008] For molecules that may potentially cause systemic toxicity (such as certain T-cell co-stimulatory molecules), FAPs can serve as "localization" targets, allowing the activity of such antitumor molecules to be confined to the local tumor environment. Therefore, there is a need to develop an FAP-targeting molecule that can act as a targeting component and selectively deliver cancer therapeutics to cancerous tissue. Summary of the Invention

[0009] Based on the disclosures provided herein, those skilled in the art will recognize, or be able to determine, many equivalents of the specific embodiments of the invention described herein using only conventional experiments. Such equivalents are intended to be covered by the following embodiments (E).

[0010] The use of section headings in this document is solely for ease of reading and is not intended to be restrictive. The entire document is intended to be viewed as a uniform disclosure, and it should be understood that all combinations of the features described herein are contemplated.

[0011] E1. A fibroblast activator protein α (FAP) antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein binds to an epitope comprising residues V275, R175, F181, Q182, I183, D178, F185, P179 and Y274 as numbered according to SEQ ID NO:394.

[0012] E2. The FAP antigen-binding protein as described in E1, wherein the epitope further comprises one or more residues selected from the group consisting of: Q174, W327, P277 and Q278 according to SEQ ID NO:394.

[0013] E3. The FAP antigen-binding protein as described in E1 or E2, wherein the epitope further comprises residues Q174, W327, P277 and Q278 numbered according to SEQ ID NO:394.

[0014] E4. The FAP antigen-binding protein as described in any one of E1-E3, wherein the epitope further comprises one or more residues selected from the group consisting of: P272, P180, I267, G276, D326 and A273 according to the numbers of SEQ ID NO:394.

[0015] E5. The FAP antigen-binding protein as described in any one of E1-E3, wherein the epitope further comprises: P272, P180, I267, G276, D326 and A273 according to the numbers of SEQ ID NO:394.

[0016] E6. The FAP antigen-binding protein as described in any one of E1-E4, wherein one or more of the following substitutions substantially disrupt the binding of the antigen-binding protein to the epitope: (1) R175 is replaced by A, N, D, C, Q, E, G, I, L, M, F, P, S, T, W, T, or V; (2) D178 is replaced by R, K, A, N, C, Q, G, I, L, M, F, P, S, T, W, Y, or V; (3) P179 is replaced by R, K, N, Q, I, M, F, W, Y, D, E, or H; (4) F181 is replaced by R, K, A, N, C, D, E, Q, G, I, L, M, P, S, T, V, or H; (5) F185 is replaced with R, K, A, N, C, D, E, Q, G, I, L, M, P, S, T, V or H; (6) Y274 is replaced with R, K, A, G, I, L, M, P, S, T, V, H, F or W; or (7) V275 is replaced with R, K, N, Q, I, L, M, H, F, W, Y, D or E.

[0017] E7. The FAP antigen-binding protein as described in E6, wherein one or more of the following substitutions substantially disrupt the binding of the protein to the epitope: (8) Q174 is replaced by A, C, G, I, L, M, F, P, W, Y, or V; (9) P277 is replaced by R, K, N, Q, I, L, M, H, F, W, Y, D, or E; or (10) Q278 is replaced by A, R, K, I, L, M, F, W, or Y.

[0018] E8. The FAP antigen-binding protein as described in E6 or E7, wherein one or more of the following substitutions substantially disrupt the binding of the protein to the epitope: (11) A273 is replaced by R, K, N, Q, I, M, F, W, Y, D, E or H; or (12) G276 is replaced by R, K, N, Q, I, L, M, H, F, W, Y, D or E.

[0019] E9. The FAP antigen-binding protein as described in any one of E1-E8, wherein the binding affinity (KD) value of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the antigen-binding protein to an epitope comprising one or more of the following substitutions: (1) R175 is replaced by A, N, D, C, Q, E, G, I, L, M, F, P, S, T, W, T, or V; (2) D178 is replaced by R, K, A, N, C, Q, G, I, L, M, F, P, S, T, W, Y, or V; (3) P179 is replaced with R, K, N, Q, I, M, F, W, Y, D, E or H; (4) F181 is replaced with R, K, A, N, C, D, E, Q, G, I, L, M, P, S, T, V or H; (5) F185 is replaced with R, K, A, N, C, D, E, Q, G, I, L, M, P, S, T, V or H; (6) Y274 is replaced with R, K, A, G, I, L, M, P, S, T, V, H, F or W; or (7) V275 is replaced with R, K, N, Q, I, L, M, H, F, W, Y, D or E.

[0020] E10. The FAP antigen-binding protein as described in E9, wherein the binding affinity (KD) value of the protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the protein to an epitope comprising one or more of the following substitutions: (8) Q174 is replaced by A, C, G, I, L, M, F, P, W, Y, or V; (9) P277 is replaced by R, K, N, Q, I, L, M, H, F, W, Y, D, or E; or (10) Q278 is replaced by A, R, K, I, L, M, F, W, or Y.

[0021] E11. The FAP antigen-binding protein as described in E9 or E10, wherein the binding affinity (KD) value of the protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the protein to an epitope comprising one or more of the following substitutions: (11) A273 is replaced by R, K, N, Q, I, M, F, W, Y, D, E, or H; or (12) G276 is replaced by R, K, N, Q, I, L, M, H, F, W, Y, D, or E.

[0022] E12. The FAP antigen-binding protein as described in any one of E9-E11, wherein the KD value is optionally measured using a Biacore T200 instrument by surface plasmon resonance (SPR).

[0023] E13. The FAP antigen-binding protein as described in any one of E9-E11, wherein the KD value is optionally measured using a ForteBio Octet instrument by biological layer interferometry (BLI).

[0024] E14. The FAP antigen-binding protein as described in any one of E1-E13, wherein the FAP is human FAP.

[0025] E15. The FAP antigen-binding protein as described in E14, wherein the FAP comprises SEQ ID NO:395.

[0026] E16. The FAP antigen-binding protein as described in any one of E1-E15, wherein the epitope is determined by X-ray crystallography or cryo-electron microscopy.

[0027] E17. The FAP antigen-binding protein as described in any one of E1-E16, wherein the VH and VL comprise (VH and VL are numbered according to Kabat): (1) H33 is Arg, Lys, Gln, or Asn; (2) H94 is Arg, Lys, Gln, or Asn; (3) H97 is Gly or Ala; (4) H98 is Tyr, Trp, Phe, Thr, or Ser; (5) H100B is Tyr, Trp, Phe, Thr, or Ser; (6) H100C is Tyr, Trp, Phe, Thr, or Ser; (7) L53 is Gln, Asn, or Glu; (8) L54 is Arg, Lys, Gln, or Asn; and (9) L60 is Asp, Glu, or Asn.

[0028] E18. The FAP antigen-binding protein as described in any one of E1-E17, wherein the VH and VL comprise (VH and VL are numbered according to Kabat): (1) H33 is Arg or Lys; (2) H94 is Arg or Lys; (3) H97 is Gly or Ala; (4) H98 is Tyr or Phe; (5) H100B is Tyr or Phe; (6) H100C is Tyr or Phe; (7) L53 is Gln or Asn; (8) L54 is Arg or Lys; and (9) L60 is Asp or Glu.

[0029] E19. The FAP antigen-binding protein as described in any one of E1-E18, wherein the VH and the VL comprise (VH and VL are numbered according to Kabat): (1) H33 is Arg; (2) H94 is Arg; (3) H97 is Gly; (4) H98 is Tyr; (5) H100B is Tyr; (6) H100C is Tyr; (7) L53 is Gln; (8) L54 is Arg; and (9) L60 is Asp.

[0030] E20. The FAP antigen-binding protein as described in any one of E17-E19, wherein the VH and VL further comprise (VH and VL are numbered according to Kabat): (10) H31 is Asn, Gln, His, Asp, Lys, or Arg; (11) H34 is Val, Ile, Leu, Met, Phe, Ala, or leucine; (12) L49 is Tyr, Trp, Phe, Thr, or Ser; and (13) L50 is Ser or Thr.

[0031] E21. The FAP antigen-binding protein as described in any one of E17-E20, wherein the VH and the VL further comprise (VH and VL are numbered according to Kabat): (10) H31 is Asn or Gln; (11) H34 is Val or Leu; (12) L49 is Tyr or Phe; and (13) L50 is Ser or Thr.

[0032] E22. The FAP antigen-binding protein as described in any one of E17-E21, wherein the VH and the VL further comprise (VH and VL are numbered according to Kabat): (10) H31 is Asn; (11) H34 is Val; (12) L49 is Tyr; and (13) L50 is Ser.

[0033] E23. The FAP antigen-binding protein as described in any one of E17-E19, wherein the VH and the VL further comprise (VH and VL are numbered according to Kabat): (14) H27 is Phe, Leu, Val, Ile, Ala or Tyr; (15) H28 is Ser or Thr; (16) H30 is Ser or Thr; (17) H95 is Ile, Leu, Val, Met, Ala, Phe or leucine; (18) H96 is Gly or Ala; (19) H101 is Asp, Glu or Asn; and (20) L52 is Asn, Gln, His, Asp, Lys or Arg.

[0034] E24. The FAP antigen-binding protein as described in any one of E17-E20, wherein the VH and VL further comprise (VH and VL are numbered according to Kabat): (14) H27 is Phe or Tyr; (15) H28 is Ser or Thr; (16) H30 is Ser or Thr; (17) H95 is Ile or Leu; (18) H96 is Gly or Ala; (19) H101 is Asp or Glu; and (20) L52 is Asn or Gln.

[0035] E25. The FAP antigen-binding protein as described in any one of E17-E21, wherein the VH and the VL further comprise (VH and VL are numbered according to Kabat): (14) H27 is Phe; (15) H28 is Ser; (16) H30 is Ser; (17) H95 is Ile; (18) H96 is Gly; (19) H101 is Asp; and (20) L52 is Asn.

[0036] E26. The FAP antigen-binding protein as described in any one of E1-E24, wherein: (a) The VH comprises: (i) a complementary determination region (CDR)-H1 comprising any one of SEQ ID NO: 398-402; and (ii) a CDR-H3 comprising any one of SEQ ID NO: 408-412; (b) The VL comprises a CDR-L2 containing any one of SEQ ID NO: 418-422; and (c) The VL is further included in the D at position L60 (according to the Kabat number).

[0037] E27. The FAP antigen-binding protein as described in E26, wherein the CDR-H1, the CDR-H3, the CDR-L2, and the residue D at L60 are selected from one or more FAP residues of the group consisting of: Q174, R175, D178, P179, P180, F181, Q182, I183, F185, I267, P272, A273, Y274, V275, G276, P277, Q278, D326, and W327 (according to SEQ ID NO:394).

[0038] E28. The FAP antigen-binding protein as described in E27, wherein the contact is defined as a distance of 4.5 Å between the heavy atoms in the FAP and the heavy atoms in the FAP antigen-binding protein, as determined by X-ray crystallography or cryo-electron microscopy.

[0039] E29. A fibroblast activator protein α (FAP) antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein binds to epitopes comprising E325, D331, Q336, I320, R324, E302, R303 and T335 according to SEQ ID NO:394.

[0040] E30. The FAP antigen-binding protein as described in E29, wherein the epitope further comprises one or more residues selected from the group consisting of K381, D322, P333 and F357 according to SEQ ID NO:394.

[0041] E31. The FAP antigen-binding protein as described in E29 or E30, wherein the epitope further comprises residues K381, D322, P333 and F357 numbered according to SEQ ID NO:394.

[0042] E32. The FAP antigen-binding protein as described in any one of E20-E31, wherein the epitope further comprises one or more residues selected from the group consisting of: K219, D326, W327, A361, I362, S363, Y364, I380, D382, F323 and H338 according to the numbers of SEQ ID NO:394.

[0043] E33. The FAP antigen-binding protein as described in any one of E29-E32, wherein the epitope further comprises residues K219, D326, W327, A361, I362, S363, Y364, I380, D382, F323 and H338 numbered according to SEQ ID NO:394.

[0044] E34. The FAP antigen-binding protein as described in any one of E29-E33, wherein one or more of the following substitutions substantially disrupt the binding of the protein to the epitope: (1) E302 is replaced by A; (2) R303 is replaced by A; (3) I320 is replaced by A; (4) R324 is replaced by A; (5) E325 is replaced by A; (6) D331 is replaced by A; (7) T335 is replaced by A; or (8) Q336 is replaced by A.

[0045] E35. The FAP antigen-binding protein as described in E34, wherein one or more of the following substitutions substantially disrupt the binding of the protein to the epitope: (9) D322 is replaced with A; or (10) P333 is replaced with W, R, K, F, H, I, M or Y; F357 is replaced with A.

[0046] E36. The FAP antigen-binding protein as described in E34 or E35, wherein one or more of the following substitutions substantially disrupt the binding of the protein to the epitope: (11) Y364 is replaced by A; or (12) D382 is replaced by W, R, K, F, H, I, M or Y.

[0047] E37. The FAP antigen-binding protein as described in any one of E29-E36, wherein the binding affinity (KD) of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD of the antigen-binding protein to an epitope comprising one or more of the following substitutions: (1) E302 is replaced with A; (2) R303 is replaced with A; (3) I320 is replaced with A; (4) R324 is replaced with A; (5) E325 is replaced with A; (6) D331 is replaced with A; (7) T335 is replaced with A; or (8) Q336 is replaced with A.

[0048] E38. The FAP antigen-binding protein as described in E37, wherein the binding affinity (KD) of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD of the antigen-binding protein to an epitope comprising one or more of the following substitutions: (9) D322 is replaced with A; or (10) P333 is replaced with W, R, K, F, H, I, M, or Y; and F357 is replaced with A.

[0049] E39. The FAP antigen-binding protein as described in E37 or E38, wherein the binding affinity (KD) value of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the antigen-binding protein to an epitope comprising one or more of the following substitutions: (11) Y364 is replaced by A; or (12) D382 is replaced by W, R, K, F, H, I, M, or Y.

[0050] E40. The FAP antigen-binding protein as described in any one of E37-E39, wherein the KD value is optionally measured using a Biacore T200 instrument by surface plasmon resonance (SPR).

[0051] E41. The FAP antigen-binding protein as described in any one of E37-E39, wherein the KD value is optionally measured using a ForteBio Octet instrument by biological layer interferometry (BLI).

[0052] E42. The FAP antigen-binding protein as described in any one of E29-E41, wherein the FAP is human FAP.

[0053] E43. The FAP antigen-binding protein as described in E42, wherein the FAP comprises SEQ ID NO:395.

[0054] E44. The FAP antigen-binding protein as described in any one of E29-E43, wherein the epitope is determined by X-ray crystallography or cryo-electron microscopy.

[0055] E45. The FAP antigen-binding protein as described in any one of E29-E44, wherein the VH comprises (VH numbered according to Kabat): (1) H31 is Arg, Lys, Gln or Asn; (2) H98 is Tyr, Trp, Phe, Thr or Ser; (3) H99 is Tyr, Trp, Phe, Thr or Ser; (4) H100 is Tyr, Trp, Phe, Thr or Ser; and (5) H101 is Asp, Glu or Asn.

[0056] E46. The FAP antigen-binding protein as described in any one of E29-E45, wherein the VH comprises (VH numbered according to Kabat): (1) H31 is Arg or Lys; (2) H98 is Tyr or Phe; (3) H99 is Tyr or Phe; (4) H100 is Tyr or Phe; and (5) H101 is Asp or Glu.

[0057] E47. The FAP antigen-binding protein as described in any one of E29-E46, wherein the VH comprises (VH numbered according to Kabat): (1) H31 is Arg; (2) H98 is Tyr; (3) H99 is Tyr; (4) H100 is Tyr; and (5) H101 is Asp.

[0058] E48. The FAP antigen-binding protein as described in any one of E45-E47, wherein the VH comprises (VH number according to Kabat): (6) H28 is Thr or Ser; and (7) H100A is Tyr, Trp, Phe, Thr or Ser.

[0059] E49. The FAP antigen-binding protein as described in any one of E45-E48, wherein the VH comprises (VH number according to Kabat): (6) H28 is Thr or Ser; and (7) H100A is Tyr or Phe.

[0060] E50. The FAP antigen-binding protein as described in any one of E45-E49, wherein the VH comprises (VH numbered according to Kabat): (6) H28 is Thr; and (7) H100A is Tyr.

[0061] E51. The FAP antigen-binding protein as described in any one of E45-E50, wherein the VH comprises (VH and VL are numbered according to Kabat): (8) H32 is Tyr, Trp, Phe, Thr, or Ser; (9) H96 is Pro or Ala; (10) H97 is Ser or Thr; (11) H100C is Tyr, Trp, Phe, Thr, or Ser; (12) L32 is Phe, Leu, Val, Ile, Ala, or Tyr; and (13) L49 is Tyr, Trp, Phe, Thr, or Ser.

[0062] E52. The FAP antigen-binding protein as described in any one of E45-E51, wherein the VH comprises (VH and VL are numbered according to Kabat): (8) H32 is Tyr or Phe; (9) H96 is Pro or Ala; (10) H97 is Ser or Thr; (11) H100C is Tyr or Phe; (12) L32 is Phe or Tyr; and (13) L49 is Tyr or Phe.

[0063] E53. The FAP antigen-binding protein as described in any one of E45-E52, wherein the VH comprises (VH and VL are numbered according to Kabat): (8) H32 is Tyr; (9) H96 is Pro; (10) H97 is Ser; (11) H100C is Tyr; (12) L32 is Phe; and (13) L49 is Tyr.

[0064] E54. The FAP antigen-binding protein as described in any one of E29-E53, wherein: (a) The VH comprises: (i) a complementary determination region (CDR)-H1 comprising any one of SEQ ID NO: 428-432; and (ii) a CDR-H3 comprising any one of SEQ ID NO: 438-442; and (b) The VL comprises a CDR-L2 containing any one of SEQ ID NO: 448-452.

[0065] E55. The FAP antigen-binding protein as described in E54, wherein the CDR-H1, the CDR-H3, and the CDR-L2 are contacted with one or more FAP residues selected from the group consisting of: K219, E302, R303, I320, D322, F323, R324, E325, D326, W327, D331, P333, T335, Q336, H338, F357, A361, I362, S363, Y364, I380, K381, and D382 (according to SEQ ID NO:394).

[0066] E56. The FAP antigen-binding protein as described in E55, wherein the contact is defined as a distance of 4.5 Å between the heavy atoms in the FAP and the heavy atoms in the FAP antigen-binding protein, as determined by X-ray crystallography or cryo-electron microscopy.

[0067] E57. A fibroblast activator protein α (FAP) antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein binds to epitopes comprising S86, E82, I181, T83, Q85, Q65, K486, N80, T88, I485 and I487 according to SEQ ID NO:394.

[0068] E58. The FAP antigen-binding protein as described in E57, wherein the epitope further comprises one or more residues selected from the group consisting of: V77, G84, Y79, and Y87 according to SEQ ID NO:394.

[0069] E59. The FAP antigen-binding protein as described in E57 or E58, wherein the epitope further comprises residues V77, G84, Y79, and Y87 numbered according to SEQ ID NO:394.

[0070] E60. The FAP antigen-binding protein as described in any one of E57-E59, wherein the epitope further comprises one or more residues selected from the group consisting of: E66, Y67, S71, D73, N75, S136 and L488 as numbered according to SEQ ID NO:394.

[0071] E61. The FAP antigen-binding protein as described in any one of E57-E60, wherein the epitope further comprises residues E66, Y67, S71, D73, N75, S136 and L488 numbered according to SEQ ID NO:394.

[0072] E62. The FAP antigen-binding protein as described in any one of E57-E61, wherein one or more of the following substitutions substantially disrupt the binding of the protein to the epitope: (1) Q65 is replaced with A; (2) N80 is replaced with A; (3) E82 is replaced with A; (4) T83 is replaced with A; (5) Q85 is replaced with A; (6) S86 is replaced with A; (7) T88 is replaced with A; (8) I485 is replaced with A; or (9) K486 is replaced with A.

[0073] E63. The FAP antigen-binding protein as described in E62, wherein one or more of the following substitutions substantially disrupt the binding of the protein to the epitope: (10) V77 is replaced by A; (11) Y79 is replaced by A; (12) G84 is replaced by A; or (13) Y87 is replaced by A.

[0074] E64. The FAP antigen-binding protein as described in E62 or E63, wherein the following substitution substantially disrupts the binding of the protein to the epitope: (14) N75 is replaced by F, Y or W.

[0075] E65. The FAP antigen-binding protein as described in any one of E57-E64, wherein the binding affinity (KD) value of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the antigen-binding protein to an epitope comprising one or more of the following substitutions: (1) Q65 is replaced with A; (2) N80 is replaced with A; (3) E82 is replaced with A; (4) T83 is replaced with A; (5) Q85 is replaced with A; (6) S86 is replaced with A; (7) T88 is replaced with A; (8) I485 is replaced with A; or (9) K486 is replaced with A.

[0076] E66. The FAP antigen-binding protein as described in E65, wherein the binding affinity (KD) value of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the antigen-binding protein to an epitope comprising one or more of the following substitutions: (10) V77 is replaced with A; (11) Y79 is replaced with A; (12) G84 is replaced with A; or (13) Y87 is replaced with A.

[0077] E67. The FAP antigen-binding protein as described in E65 or E66, wherein the binding affinity (KD) value of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the antigen-binding protein to an epitope comprising the following substitutions: (14) N75 is replaced by F, Y, or W.

[0078] E68. The FAP antigen-binding protein as described in any one of E65-E67, wherein the KD value is optionally measured using a Biacore T200 instrument by surface plasmon resonance (SPR).

[0079] E69. The FAP antigen-binding protein as described in any one of E65-E67, wherein the KD value is optionally measured using a ForteBio Octet instrument by biological layer interferometry (BLI).

[0080] E70. The FAP antigen-binding protein as described in any one of E57-E69, wherein the FAP is human FAP.

[0081] E71. The FAP antigen-binding protein as described in E70, wherein the FAP comprises SEQ ID NO:395.

[0082] E72. The FAP antigen-binding protein as described in any one of E57-E71, wherein the epitope is determined by X-ray crystallography or cryo-electron microscopy.

[0083] E73. The FAP antigen-binding protein as described in any one of E57-E72, wherein the VH comprises (VH numbered according to Kabat): (1) H31 is Asn, Gln, His, Asp, Lys, or Arg; (2) H33 is Gly or Ala; (3) H52A is Tyr, Trp, Phe, Thr, or Ser; (4) H55 is Arg, Lys, Gln, or Asn; (5) H56 is Asn, Gln, His, Asp, Lys, or Arg; (6) H95 is Asp, Glu, or Asn; (7) H100 is Gly or Ala; (8) L32 is Tyr, Trp, Phe, Thr, or Ser; (9) L91 is Phe, Leu, Val, Ile, Ala, or Tyr; and (10) L95 is Tyr, Trp, Phe, Thr, or Ser.

[0084] E74. The FAP antigen-binding protein as described in any one of E57-E73, wherein the VH comprises (VH numbered according to Kabat): (1) H31 is Asn or Gln; (2) H33 is Gly or Ala; (3) H52A is Tyr or Phe; (4) H55 is Arg or Lys; (5) H56 is Asn or Gln; (6) H95 is Asp or Glu; (7) H100 is Gly or Ala; (8) L32 is Tyr or Phe; (9) L91 is Phe or Tyr; and (10) L95 is Tyr or Phe.

[0085] E75. The FAP antigen-binding protein as described in any one of E57-E74, wherein the VH comprises (VH numbered according to Kabat): (1) H31 is Asn; (2) H33 is Gly; (3) H52A is Tyr; (4) H55 is Arg; (5) H56 is Asn; (6) H95 is Asp; (7) H100 is Gly; (8) L32 is Tyr; (9) L91 is Phe; and (10) L95 is Tyr.

[0086] E76. The FAP antigen-binding protein as described in any one of E73-E75, wherein the VH comprises (VH numbered according to Kabat): (11) H28 is Thr or Ser; (12) H32 is Tyr, Trp, Phe, Thr or Ser; (13) H52 is Trp, Tyr or Phe; (14) L30 is Tyr, Trp, Phe, Thr or Ser; and (15) L96 is Trp, Tyr or Phe.

[0087] E77. The FAP antigen-binding protein as described in any one of E73-E76, wherein the VH comprises (VH numbered according to Kabat): (11) H28 is Thr or Ser; (12) H32 is Tyr or Phe; (13) H52 is Trp or Tyr; (14) L30 is Tyr or Phe; and (15) L96 is Trp or Tyr.

[0088] E78. The FAP antigen-binding protein as described in any one of E73-E77, wherein the VH comprises (VH numbered according to Kabat): (11) H28 is Thr; (12) H32 is Tyr; (13) H52 is Trp; (14) L30 is Tyr; and (15) L96 is Trp.

[0089] E79. The FAP antigen-binding protein as described in any one of E73-E78, wherein the VH comprises (VH and VL are numbered according to Kabat): (16) H30 is Asn, Gln, His, Asp, Lys or Arg; (17) H53 is Asp, Glu or Asn; (18) H96 is Gly or Ala; (19) H97 is Ser or Thr; (20) H98 is Gly or Ala; (21) H99 is Gly or Ala; and (22) L56 is Ser or Thr.

[0090] E80. The FAP antigen-binding protein as described in any one of E73-E79, wherein the VH comprises (VH and VL are numbered according to Kabat): (16) H30 is Asn or Gln; (17) H53 is Asp or Glu; (18) H96 is Gly or Ala; (19) H97 is Ser or Thr; (20) H98 is Gly or Ala; (21) H99 is Gly or Ala; and (22) L56 is Ser or Thr.

[0091] E81. The FAP antigen-binding protein as described in any one of E73-E80, wherein the VH comprises (VH and VL are numbered according to Kabat): (16) H30 is Asn; (17) H53 is Asp; (18) H96 is Gly; (19) H97 is Ser; (20) H98 is Gly; (21) H99 is Gly; and (22) L56 is Ser.

[0092] E82. The FAP antigen-binding protein as described in any one of E57-E81, wherein: (a) The VH comprises: (i) a CDR-H1 comprising any one of SEQ ID NO: 458-462; (ii) a CDR-H2 comprising any one of SEQ ID NO: 463-467; and (iii) a CDR-H3 comprising any one of SEQ ID NO: 468-472; and (b) The VL comprises: (i) a CDR-L1 comprising any one of SEQ ID NO: 473-487; (ii) a CDR-L2 comprising any one of SEQ ID NO: 478-482; and (iii) a CDR-L3 comprising any one of SEQ ID NO: 483-487.

[0093] E83. The FAP antigen-binding protein as described in E45, wherein the CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 contact one or more FAP residues selected from the group consisting of: Q65, N75, V77, Y79, N80, E82, T83, G84, Q85, S86, Y87, T88, I485 and K486 (according to SEQ ID NO:394).

[0094] E84. The FAP antigen-binding protein as described in E83, wherein the contact is defined as a distance of 4.5 Å between the heavy atoms in the FAP and the heavy atoms in the FAP antigen-binding protein, as determined by X-ray crystallography or cryo-electron microscopy.

[0095] E85. A fibroblast activator protein α (FAP) antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein binds to epitopes comprising K381, K371, E414, S428, Q389, Y432, I390, P434 and K436 according to SEQ ID NO:394.

[0096] E86. The FAP antigen-binding protein as described in E85, wherein the epitope further comprises one or more residues selected from the group consisting of: I427, I388, G430, P433 and I426 according to SEQ ID NO:394.

[0097] E87. The FAP antigen-binding protein as described in E85 or E86, wherein the epitope further comprises residues I427, I388, G430, P433 and I426 numbered according to SEQ ID NO:394.

[0098] E88. The FAP antigen-binding protein as described in any one of E85-E87, wherein the epitope further comprises one or more residues selected from the group consisting of: W395, S435, Y379, N386, A387 and S392 according to SEQ ID NO:394.

[0099] E89. The FAP antigen-binding protein as described in any one of E85-E88, wherein the epitope further comprises W395, S435, Y379, N386, A387 and S392 according to the numbers of SEQ ID NO:394.

[0100] E90. The FAP antigen-binding protein as described in any one of E85-E89, wherein one or more of the following substitutions substantially disrupt the binding of the antigen-binding protein to the epitope: (1) K371 is replaced by A; (2) K381 is replaced by A; (3) E414 is replaced by A; (4) I390 is replaced by A; (5) Y432 is replaced by A; (6) P434 is replaced by F; or (7) K436 is replaced by A.

[0101] E91. The FAP antigen-binding protein as described in E90, wherein one or more of the following substitutions substantially disrupt the binding of the antigen-binding protein to the epitope: (8) I388 is replaced by A; (9) I427 is replaced by A; (10) G430 is replaced by A; or (11) P433 is replaced by F.

[0102] E92. The FAP antigen-binding protein as described in any one of E85-E91, wherein the binding affinity (KD) value of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the antigen-binding protein to an epitope comprising one or more of the following substitutions: (1) K371 is replaced with A; (2) K381 is replaced with A; (3) E414 is replaced with A; (4) I390 is replaced with A; (5) Y432 is replaced with A; (6) P434 is replaced with F; or (7) K436 is replaced with A.

[0103] E93. The FAP antigen-binding protein as described in E92, wherein the binding affinity (KD) value of the antigen-binding protein to the epitope is at least 100 times, at least 200 times, at least 300 times, at least 400 times, at least 500 times, at least 600 times, at least 700 times, at least 800 times, at least 900 times, or at least 1000 times lower than the KD value of the antigen-binding protein to an epitope comprising one or more of the following substitutions: (8) I388 is replaced with A; (9) I427 is replaced with A; (10) G430 is replaced with A; or (11) P433 is replaced with F.

[0104] E94. FAP antigen-binding protein as described in E92 or E93, wherein the KD value is optionally measured using a Biacore T200 instrument via surface plasmon resonance (SPR).

[0105] E95. FAP antigen-binding protein as described in E92 or E93, wherein the KD value is optionally measured using a ForteBio Octet instrument via biological layer interferometry (BLI).

[0106] E96. The FAP antigen-binding protein as described in any one of E85-E95, wherein the FAP is human FAP.

[0107] E97. The FAP antigen-binding protein as described in E96, wherein the FAP comprises SEQ ID NO:395.

[0108] E98. The FAP antigen-binding protein as described in any one of E85-E97, wherein the epitope is determined by X-ray crystallography or cryo-electron microscopy.

[0109] E99. The FAP antigen-binding protein as described in any one of E85-E98, wherein the VH and VL comprise (VH and VL are numbered according to Kabat): (1) H28 is Thr or Ser; (2) H53 is Asp, Glu, or Asn; (3) H96 is Arg, Lys, Gln, or Asn; (4) H99 is Tyr, Trp, Phe, Thr or Ser; (5) H100A is Tyr, Trp, Phe, Thr or Ser; (6) H100B is Tyr, Trp, Phe, Thr or Ser; and (7) H100C is Tyr, Trp, Phe, Thr or Ser.

[0110] E100. The FAP antigen-binding protein as described in any one of E85-E99, wherein the VH and VL comprise (VH and VL are numbered according to Kabat): (1) H28 is Thr or Ser; (2) H53 is Asp or Glu; (3) H96 is Arg or Lys; (4) H99 is Tyr or Phe; (5) H100A is Tyr or Phe; (6) H100B is Tyr or Phe; and (7) H100C is Tyr or Phe.

[0111] E101. The FAP antigen-binding protein as described in any one of E85-E100, wherein the VH and the VL comprise (VH and VL are numbered according to Kabat): (1) H28 is Thr; (2) H53 is Asp; (3) H96 is Arg; (4) H99 is Tyr; (5) H100A is Tyr; (6) H100B is Tyr; and (7) H100C is Tyr.

[0112] E102. The FAP antigen-binding protein as described in any one of E99-E101, wherein the VH and VL comprise (VH and VL are numbered according to Kabat): (8) H31 is Ser or Thr; (9) H52 is Trp, Tyr, or Phe; (10) H97 is Leu, leucine, Ile, Val, Met, Ala, or Phe; (11) H98 is Gln, Asn, or Glu; (12) L32 is Leu, leucine, Ile, Val, Met, Ala, or Phe; (13) L91 is Tyr, Trp, Phe, Thr, or Ser; and (14) L96 is Trp, Tyr, or Phe.

[0113] E103. The FAP antigen-binding protein as described in any one of E99-E102, wherein the VH and the VL comprise (VH and VL numbered according to Kabat): (8) H31 is Ser or Thr; (9) H52 is Trp or Tyr; (10) H97 is Leu or Ile; (11) H98 is Gln or Asn; (12) L32 is Leu or Ile; (13) L91 is Tyr or Phe; and (14) L96 is Trp or Tyr.

[0114] E104. The FAP antigen-binding protein as described in any one of E99-E103, wherein the VH and the VL comprise (VH and VL are numbered according to Kabat): (8) H31 is Ser; (9) H52 is Trp; (10) H97 is Leu; (11) H98 is Gln; (12) L32 is Leu; (13) L91 is Tyr; and (14) L96 is Trp.

[0115] E105. The FAP antigen-binding protein as described in any one of E99-E104, wherein the VH and the VL comprise (VH and VL are numbered according to Kabat): (15) H30 is Ser or Thr; (16) H95 is Asp, Glu, or Asn; (17) H100 is Asp, Glu, or Asn; and (18) L30 is Tyr, Trp, Phe, Thr, or Ser.

[0116] E106. The FAP antigen-binding protein as described in any one of E99-E105, wherein the VH and the VL comprise (VH and VL are numbered according to Kabat): (15) H30 is Ser or Thr; (16) H95 is Asp and Glu; (17) H100 is Asp or Glu; and (18) L30 is Tyr and Phe.

[0117] E107. The FAP antigen-binding protein as described in any one of E99-E106, wherein the VH and the VL comprise (VH and VL are numbered according to Kabat): (15) H30 is Ser; (16) H95 is Asp; (17) H100 is Asp; and (18) L30 is Tyr.

[0118] E108. The FAP antigen-binding protein as described in any one of E85-E107, wherein: (a) The VH comprises: (i) a CDR-H1 comprising any one of SEQ ID NO: 488-492; (ii) a CDR-H2 comprising any one of SEQ ID NO: 493-497; and (iii) a CDR-H3 comprising any one of SEQ ID NO: 498-502; and (b) The VL comprises: (i) a CDR-L1 comprising any one of SEQ ID NO: 503-507; and (ii) a CDR-L3 comprising any one of SEQ ID NO: 513-517.

[0119] E109. The FAP antigen-binding protein as described in E108, wherein the CDR-H1, CDR-H2, CDR-H3, CDR-L2, and CDR-L3 contact one or more FAP residues selected from the group consisting of: K381, K371, E414, S428, Q389, Y432, I390, P434, K436, I427, I388, G430, P433, I429, W395, S435, Y379, N386, A387, and S392 (according to SEQ ID NO:394).

[0120] E110. The FAP antigen-binding protein as described in E109, wherein the contact is defined as a distance of 4.5 Å between the heavy atoms in the FAP and the heavy atoms in the FAP antigen-binding protein, as determined by X-ray crystallography or cryo-electron microscopy.

[0121] E111. A FAP antigen-binding protein, the FAP antigen-binding protein comprising: (1) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 197, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 198; (2) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 199, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 200; (3) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 201, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 202; (4) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 203, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 204; (5) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 205, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 206; (6) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 207, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 208; (7) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 209, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 210; (8) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 211, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 212; (9) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 213, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 214; (10) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 215, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 216; (11) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 217, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 218; (12) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 219, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 220; (13) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 221, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 222; (14) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 223, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 224; (15) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 225, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 226; (16) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 227, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 228; (17) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 229, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 230; (18) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 231, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 232; (19) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 233, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 234; (20) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 235, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 236; (21) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 237, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 238; (22) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 239, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 240; (23) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 241, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 242; (24) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 243, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 244; (25) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 245, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 246; (26) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 247, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 248; (27) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 249, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 250; (28) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 251, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 252; (29) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 253, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 254; (30) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 255, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 256; (31) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 257, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 258; (32) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 259, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 260; (33) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 261, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 262; (34) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 263, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 264; (35) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 265, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 266; (36) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 267, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 268; (37) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 269, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 270; (38) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 271, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 272; (39) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 273, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 274; (40) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 275, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 276; (41) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 277, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 278; (42) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 279, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 280; (43) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 281, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 282; (44) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 283, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 284; (45) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 285, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 286; (46) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 287, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 288; (47) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 289, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 290; (48) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 291, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 292; (49) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 293, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 294; (50) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 295, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 296; (51) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 297, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 298; (52) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 299, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 300; (53) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 301, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 302; (54) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 303, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 304; (55) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 305, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 306; (56) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 307, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 308; (57) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 309, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 310; (58) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 311, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 312; (59) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 313, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 314; (60) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 315, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 316; (61) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 317, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 318; (62) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 319, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 320; (63) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 321, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 322; (64) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 323, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 324; (65) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 325, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 326; (66) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 327, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 328; (67) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 329, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 330; (68) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 331, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 332; (69) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 333, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 334; (70) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 335, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 336; (71) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 337, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 338; (72) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 339, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 340; or (73) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 341, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 342.

[0122] E112. An FAP antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the VH and the VL comprise: (i) CDR-H1, which contains the same information as SEQ ID NO: 1, 7, 13, 19, 25, 31, 37, 43, 49, 55, 61, 67, 73, 79, 85, 91, 97, 103, 109, 115, 121, 127, 133, 139, 145, 151, 157, 163, 169, 175, 181, 187, 398, 399, 400, 401, 402, 428, 429, 430, 431, 432, 458, 459, 460, 461, 462, 488, 489, 490, 491 or 492 at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical sequences; (ii) CDR-H2, which contains the same as SEQ ID NO: 2, 8, 14, 20, 26, 32, 38, 44, 50, 56, 62, 68, 74, 80, 86, 92, 98, 104, 110, 116, 122, 128, 134, 140, 146, 152, 158, 164, 170, 176, 182, 188, 403, 404, 405, 406, 407, 433, 434, 435, 436, 437, 463, 464, 465, 466, 467, 493, 494, 495, 496 or 497 at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical sequences; (iii) CDR-H3, which contains the same as SEQ ID NO: 3, 9, 15, 21, 27, 33, 39, 45, 51, 57, 63, 69, 75, 81, 87, 93, 99, 105, 111, 117, 123, 129, 135, 141, 147, 153, 159, 165, 171, 177, 183, 189, 194, 196, 408, 409, 410, 411, 412, 438, 439, 440, 441, 442, 468, 469, 470, 471, 472, 498, 499, 500, 501 or 502 at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical sequences; (iv) CDR-L1, which contains the same information as SEQ ID NO: 4, 10, 16, 22, 28, 34, 40, 46, 52, 58, 64, 70, 76, 82, 88, 94, 100, 106, 112, 118, 124, 130, 136, 142, 148, 154, 160, 166, 172, 178, 184, 190, 413, 414, 415, 416, 417, 443, 444, 445, 446, 447, 473, 474, 475, 476, 477, 503, 504, 505, 506 or 507 at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical sequences; (v) CDR-L2, which contains SEQ ID NO: 5, 11, 17, 23, 29, 35, 41, 47, 53, 59, 65, 71, 77, 83, 89, 95, 101, 107, 113, 119, 125, 131, 137, 143, 149, 155, 161, 167, 173, 179, 185, 191, 418, 419, 420, 421, 422, 448, 449, 450, 451, 452, 478, 479, 480, 481, 482, 508, 509, 510, 511, or 512 at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical sequences; and (vi) CDR-L3, which contains SEQ ID NO: 6, 12, 18, 24, 30, 36, 42, 48, 54, 60, 66, 72, 78, 84, 90, 96, 102, 108, 114, 120, 126, 132, 138, 144, 150, 156, 162, 168, 174, 180, 186, 192, 193, 195, 423, 424, 425, 426 427, 453, 454, 455, 456, 457, 483, 484, 485, 486, 487, 513, 514, 515, 516 or 517 are at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical sequences.

[0123] E113. The FAP antigen-binding protein as described in any one of E1-E112, wherein the FAP antigen-binding protein comprises: (1) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 1-6 respectively; (2) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 6-12 respectively; (3) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 13-18; (4) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 19-24 respectively; (5) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 25-30; (6) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 31-36 respectively; (7) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 37-42; (8) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 43-48; (9) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 49-54; (10) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 55-60 respectively; (11) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 61-66; (12) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 67-72 respectively; (13) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 73-78; (14) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 79-84; (15) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 85-90; (16) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 91-96 respectively; (17) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 97-102; (18) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 103-108; (19) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 109-114; (20) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 115-120; (21) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 121-126; (22) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 127-132; (23) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 133-138; (24) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 139-144; (25) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 145-150; (26) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 151-156; (27) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 157-162; (28) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 163-168; (29) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 169-174; (30) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 175-180; (31) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 181-186; (32) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 187-192; (33) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 43, 44, 45, 46, 47 and 193; (34) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 43, 44, 194, 46, 47 and 193; (35) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 respectively containing SEQ ID NO: 175, 176, 177, 178, 179, and 195; or (36) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 187, 188, 196, 190, 191 and 192.

[0124] E114. The FAP antigen-binding protein as described in any one of E1-E112, wherein: (a) The VH comprises: (i) a CDR-H1 comprising any one of SEQ ID NO: 398-402; (ii) a CDR-H2 comprising any one of SEQ ID NO: 403-407; and (iii) a CDR-H3 comprising any one of SEQ ID NO: 408-412; and (b) The VL comprises: (i) a CDR-L1 comprising any one of SEQ ID NO: 413-417; (ii) a CDR-L2 comprising any one of SEQ ID NO: 418-422; and (iii) a CDR-L3 comprising any one of SEQ ID NO: 423-427.

[0125] E115. The FAP antigen-binding protein as described in any one of E1-E112, wherein: (a) The VH comprises: (i) a CDR-H1 comprising any one of SEQ ID NO: 428-432; (ii) a CDR-H2 comprising any one of SEQ ID NO: 433-437; and (iii) a CDR-H3 comprising any one of SEQ ID NO: 438-442; and (b) The VL comprises: (i) a CDR-L1 comprising any one of SEQ ID NO: 443-447; (ii) a CDR-L2 comprising any one of SEQ ID NO: 448-452; and (iii) a CDR-L3 comprising any one of SEQ ID NO: 453-457.

[0126] E116. The FAP antigen-binding protein as described in any one of E1-E112, wherein: (a) The VH comprises: (i) a CDR-H1 comprising any one of SEQ ID NO: 458-462; (ii) a CDR-H2 comprising any one of SEQ ID NO: 463-467; and (iii) a CDR-H3 comprising any one of SEQ ID NO: 468-472; and (b) The VL comprises: (i) a CDR-L1 comprising any one of SEQ ID NO: 473-477; (ii) a CDR-L2 comprising any one of SEQ ID NO: 478-482; and (iii) a CDR-L3 comprising any one of SEQ ID NO: 483-487.

[0127] E117. The FAP antigen-binding protein as described in any one of E1-E112, wherein: (a) The VH comprises: (i) a CDR-H1 comprising any one of SEQ ID NO: 488-492; (ii) a CDR-H2 comprising any one of SEQ ID NO: 493-497; and (iii) a CDR-H3 comprising any one of SEQ ID NO: 498-502; and (b) The VL comprises: (i) a CDR-L1 comprising any one of SEQ ID NO: 503-507; (ii) a CDR-L2 comprising any one of SEQ ID NO: 508-512; and (iii) a CDR-L3 comprising any one of SEQ ID NO: 513-517.

[0128] E118. The FAP antigen-binding protein as described in any one of E1-E117, wherein the FAP antigen-binding protein comprises a VL frame derived from a human germline Vκ frame sequence such as a Vκ1 frame sequence or a Vκ2 frame sequence.

[0129] E119. The FAP antigen-binding protein as described in any one of E1-E117, wherein the FAP antigen-binding protein comprises a VL frame derived from a human Vλ frame sequence such as a Vλ1 frame sequence, a Vλ2 frame sequence, or a Vλ3 frame sequence.

[0130] E120. The FAP antigen-binding protein as described in any one of E1-E119, wherein the FAP antigen-binding protein comprises a VH frame derived from a human VH1, VH2, VH3, VH4 or VH5 frame sequence.

[0131] E121. The FAP antigen-binding protein as described in any one of E1-E120, wherein the FAP antigen-binding protein comprises a VH frame derived from a human VH1 frame sequence.

[0132] E122. The FAP antigen-binding protein as described in any one of E1-E120, wherein the FAP antigen-binding protein comprises a VH frame derived from a human VH2 frame sequence.

[0133] E123. The FAP antigen-binding protein as described in any one of E1-E120, wherein the FAP antigen-binding protein comprises a VH frame derived from a human VH3 frame sequence.

[0134] E124. The FAP antigen-binding protein as described in any one of E1-E120, wherein the FAP antigen-binding protein comprises a VH frame derived from a human VH4 frame sequence.

[0135] E125. The FAP antigen-binding protein as described in any one of E118-E124, comprising a VL frame sequence and a VH frame sequence, wherein the VL frame sequence is at least 90% identical to the human germline frame sequence from which it is derived, and the VH frame sequence is at least 90% identical to the human germline frame sequence from which it is derived.

[0136] E126. The FAP antigen-binding protein as described in any one of E118-E125, comprising a VL frame sequence and a VH frame sequence, wherein the VL frame sequence is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a human germline frame sequence derived therefrom, and the VH frame sequence is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to a human germline frame sequence derived therefrom.

[0137] E127. The FAP antigen-binding protein as described in any one of E1-E126, wherein the FAP antigen-binding protein comprises the following: SEQ ID NO: 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287. 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 333, 335, 337, 339 or 341 at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical heavy chain variable regions (VH).

[0138] E128. The FAP antigen-binding protein as described in any one of E1-E127, wherein the FAP antigen-binding protein comprises the following: SEQ ID NO: 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288. 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 332, 334, 336, 338, 340 or 342 at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% identical light chain variable regions (VL).

[0139] E129. The FAP antigen-binding protein as described in any one of E1-E128, wherein the FAP antigen-binding protein comprises: (1) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 197, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 198; (2) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to SEQ ID NO: 199, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to SEQ ID NO: 200; (3) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 201, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 202; (4) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 203, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 204; (5) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 205, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 206; (6) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 207, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 208; (7) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 209, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 210; (8) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 211, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 212; (9) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 213, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 214; (10) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 215, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 216; (11) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 217, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 218; (12) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 219, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 220; (13) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 221, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 222; (14) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 223, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 224; (15) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 225, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 226; (16) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 227, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 228; (17) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 229, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 230; (18) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 231, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 232; (19) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 233, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 234; (20) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 235, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 236; (21) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 237, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 238; (22) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 239, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 240; (23) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 241, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 242; (24) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 243, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 244; (25) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 245, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 246; (26) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 247, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 248; (27) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 249, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 250; (28) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 251, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 252; (29) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 253, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 254; (30) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 255, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 256; (31) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 257, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 258; (32) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 259, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 260; (33) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 261, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 262; (34) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 263, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 264; (35) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 265, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 266; (36) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 267, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 268; (37) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 269, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 270; (38) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 271, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 272; (39) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 273, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 274; (40) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 275, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 276; (41) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 277, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 278; (42) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 279, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 280; (43) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 281, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 282; (44) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 283, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 284; (45) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 285, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 286; (46) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 287, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 288; (47) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 289, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 290; (48) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 291, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 292; (49) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 293, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 294; (50) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 295, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 296; (51) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 297, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 298; (52) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 299, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 300; (53) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 301, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 302; (54) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 303, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 304; (55) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 305, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 306; (56) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 307, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 308; (57) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 309, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 310; (58) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 311, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 312; (59) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 313, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 314; (60) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 315, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 316; (61) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 317, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 318; (62) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 319, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 320; (63) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 321, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 322; (64) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 323, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 324; (65) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 325, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 326; (66) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 327, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 328; (67) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 329, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 330; (68) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 331, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 332; (69) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 333, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 334; (70) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 335, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that in SEQ ID NO: 336; (71) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 337, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 338; (72) A VH comprising at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical sequences to SEQ ID NO: 339, and a VL comprising at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical sequences to SEQ ID NO: 340; or (73) A VH containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 341, and a VL containing at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% of the sequence identical to that of SEQ ID NO: 342.

[0140] E130. The FAP antigen-binding protein as described in any one of E1-E129, wherein the FAP antigen-binding protein further comprises a heavy chain CH1 domain.

[0141] E131. The FAP antigen-binding protein as described in E130, wherein the CH1 domain is the CH1 domain of IgG (e.g., IgG1, IgG2, IgG3, or IgG4).

[0142] E132. The FAP antigen-binding protein as described in E130 or E131, wherein the CH1 domain is the CH1 domain of human IgG (e.g., human IgG1, human IgG2, human IgG3, or human IgG4).

[0143] E133. The FAP antigen-binding protein as described in any one of E130-E132, wherein the CH1 domain comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO: 347, 364, 368, or 372.

[0144] E134. The FAP antigen-binding protein as described in any one of E130-E133, wherein the CH1 domain comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO:347.

[0145] E135. The FAP antigen-binding protein as described in any one of E1-E134, wherein the FAP antigen-binding protein further comprises an Fc region.

[0146] E136. The FAP antigen-binding protein as described in E135, wherein the Fc region is the Fc region of IgA (e.g., IgA1 or IgA2), IgD, IgE, IgM, or IgG (e.g., IgG1, IgG2, IgG3, or IgG4).

[0147] E137. The FAP antigen-binding protein as described in E135 or E136, wherein the Fc region is the Fc region of IgG.

[0148] E138. The FAP antigen-binding protein as described in E137, wherein the IgG is selected from the group consisting of IgG1, IgG2, IgG3 and IgG4.

[0149] E139. The FAP antigen-binding protein as described in E138, wherein the IgG is IgG1, IgG2, or IgG4.

[0150] E140. The FAP antigen-binding protein as described in any one of E135-E139, wherein the Fc region is derived from IgGFc and further comprises one or more mutations selected from the group consisting of: L234A, L235A, L235E, G237A and combinations thereof (according to EU index number).

[0151] E141. The FAP antigen-binding protein as described in E140, wherein the FAP antigen-binding protein contains L234A and L235A mutations.

[0152] E142. The FAP antigen-binding protein as described in any one of E135-E141, wherein the Fc region is derived from IgGFc and further comprises one or more mutations selected from the group consisting of: V259C, A287C, R292C, V302C, L306C, V323C, I332C and combinations thereof (according to EU index number).

[0153] E143. The FAP antigen-binding protein as described in any one of E135-E142, wherein the Fc region is derived from IgGFc and further comprises one or more mutations selected from the group consisting of: L242C, A287C, R292C, N297G, V302C, L306C, K334C and combinations thereof (according to EU index number).

[0154] E144. The FAP antigen-binding protein as described in E143, which contains the N297G mutation.

[0155] E145. The FAP antigen-binding protein as described in E143, wherein the FAP antigen-binding protein contains mutations of A287C, N297G, and L306C.

[0156] E146. The FAP antigen-binding protein as described in E143, wherein the FAP antigen-binding protein contains the R292C, N297G and V302C mutations.

[0157] E147. The FAP antigen-binding protein as described in any one of E135-E146, wherein the Fc region is derived from IgGFc and further comprises one or more mutations selected from the group consisting of M252Y, S254T, T256E and combinations thereof.

[0158] E148. The FAP antigen-binding protein as described in E147, wherein the FAP antigen-binding protein contains the M252Y, S254T, and T256E mutations.

[0159] E149. The FAP antigen-binding protein as described in any one of E135-E148, wherein the lysine residue (K) at the C-terminus of the Fc region is missing.

[0160] E150. The FAP antigen-binding protein as described in any one of E135-E148, wherein a lysine residue (K) is present at the C-terminus of the Fc region.

[0161] E151. The FAP antigen-binding protein as described in any one of E135-E148, wherein glycine and lysine residues (GK) are present at the C-terminus of the Fc region.

[0162] E152. The FAP antigen-binding protein as described in any one of E135-E148, wherein the glycine and lysine residues (GK) at the C-terminus of the Fc region are missing.

[0163] E153. The FAP antigen-binding protein as described in any one of E135-E152, wherein the Fc region comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO: 348, 355, 356, 357, 358, 359, 360, 366, 370, or 374.

[0164] E154. The FAP antigen-binding protein as described in any one of E116-E134, wherein the Fc region comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO: 348, 355, 356, 357, 358, 359, or 360.

[0165] E155. The FAP antigen-binding protein as described in any one of E1-E154, wherein the FAP antigen-binding protein further comprises a heavy chain constant domain.

[0166] E156. The FAP antigen-binding protein as described in E155, wherein the constant domain is a constant domain of IgA (e.g., IgA1 or IgA2), IgD, IgE, IgM, or IgG (e.g., IgG1, IgG2, IgG3, or IgG4).

[0167] E157. The FAP antigen-binding protein as described in E155 or E156, wherein the constant domain is a constant domain of IgG (e.g., IgG1, IgG2, IgG3, or IgG4), preferably human IgG (e.g., human IgG1, human IgG2, human IgG3, or human IgG4).

[0168] E158. The FAP antigen-binding protein as described in any one of E155-E157, wherein the constant domain comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO: 349, 350, 351, 352, 353, 354, 361, 362, 363, 367, 371, or 375.

[0169] E159. The FAP antigen-binding protein as described in any one of E155-E158, wherein the constant domain comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO: 349, 350, 351, 352, 353, 354, 361, 362, or 363.

[0170] E160. The FAP antigen-binding protein as described in any one of E1-E159, wherein the FAP antigen-binding protein further comprises a κ or λ light chain constant domain.

[0171] E161. The FAP antigen-binding protein as described in any one of E1-E160, the FAP antigen-binding protein further comprising at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the same κ light chain constant domain as SEQ ID NO:343 or 344.

[0172] E162. The FAP antigen-binding protein as described in any one of E1-E160, further comprising at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the same λ light chain constant domain as SEQ ID NO:345 or 346.

[0173] E163. The FAP antigen-binding protein as described in any one of E1-E162, wherein the FAP antigen-binding protein is an antibody.

[0174] E164. The FAP antigen-binding protein as described in any one of E1-E162, wherein the FAP antigen-binding protein is an antigen-binding fragment of an antibody, such as a Fab fragment.

[0175] E165. The FAP antigen-binding protein as described in any one of E1-E162, wherein the FAP antigen-binding protein is scFv.

[0176] E166. The FAP antigen-binding protein as described in E165, wherein the scFv contains a first linker between VH and VL.

[0177] E167. The FAP antigen-binding protein as described in E166, wherein the first linker comprises: (a) a glycine-rich peptide; (b) a peptide comprising glycine and serine; and (c) a peptide comprising (Gly-Gly-Ser). n (d) A peptide containing (Gly-Gly-Gly-Ser) where n is 1, 2, 3, 4, 5, or 6 (SEQ ID NO: 388); n (e) A peptide, wherein n is 1, 2, 3, 4, 5, or 6 (SEQ ID NO: 386); n The peptide, wherein n is 1, 2, 3, 4, 5 or 6 (SEQ ID NO:387); or (f) contains (Gly-Gly-Gly-Gly-Gln) n The peptide, wherein n is 1, 2, 3, 4, 5 or 6 (SEQ ID NO:389).

[0178] E168. The FAP antigen-binding protein as described in E166 or E167, wherein the first linker comprises the amino acid sequence (Gly-Gly-Gly-Gly-Ser)3 (SEQ ID NO:387).

[0179] E169. The FAP antigen-binding protein as described in any one of E165-E168, wherein the FAP antigen-binding protein comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO:519.

[0180] E170. The FAP antigen-binding protein as described in any one of E165-E168, the FAP antigen-binding protein comprising at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the sequence identical to that of SEQ ID NO:520.

[0181] E171. The FAP antigen-binding protein as described in any one of E165-E168, wherein the FAP antigen-binding protein comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence of SEQ ID NO:521.

[0182] E172. The FAP antigen-binding protein as described in any one of E165-E168, wherein the FAP antigen-binding protein comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO:518.

[0183] E173. The FAP antigen-binding protein as described in any one of E165-E168, the FAP antigen-binding protein comprising at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the sequence identical to that of SEQ ID NO:393.

[0184] E174. The FAP antigen-binding protein as described in any one of Examples E1-E162, wherein the FAP antigen-binding protein is Fab.

[0185] E175. The FAP antigen-binding protein as described in any one of E1-E174, wherein the antigen-binding protein has a K value equal to or less than the following: D Values ​​combined with FAP: approximately 200 nM, approximately 150 nM, approximately 100 nM, approximately 90 nM, approximately 80 nM, approximately 70 nM, approximately 60 nM, approximately 50 nM, approximately 40 nM, approximately 30 nM, approximately 25 nM, approximately 20 nM, approximately 15 nM, approximately 10 nM, approximately 9 nM, approximately 8 nM, approximately 7 nM, approximately 6 nM, approximately 5 nM, approximately 4 nM, approximately 3 nM, approximately 2 nM, approximately 1 nM, approximately 900 pM, approximately 800 pM, approximately 700 pM, approximately 600 pM, approximately 500 pM, approximately 400 pM, approximately 300 pM, approximately 250 pM, approximately 200 pM, approximately 150 pM, approximately 100 pM, approximately 50 pM, approximately 40 pM, approximately 30 pM, approximately 25 pM, approximately 20 pM, approximately 15 pM, approximately 10 pM, approximately 5 pM, or approximately 1 pM.

[0186] E176. A FAP antigen-binding protein that competitively binds to FAP with any of the FAP antigen-binding proteins described in E1-E175.

[0187] E177. An FAP antigen-binding protein that binds to an epitope substantially identical to any of the FAP antigen-binding proteins described in E1-E175.

[0188] E178. The FAP antigen-binding protein as described in any one of Examples E1-E177, wherein the FAP antigen-binding protein is a bispecific molecule further comprising a second antigen-binding moiety.

[0189] E179. A nucleic acid comprising a nucleotide sequence encoding a FAP antigen-binding protein as described in any one of E1-E178.

[0190] E180. A vector comprising nucleic acid as described in E179.

[0191] E181. A host cell comprising a nucleic acid as described in E179 or a vector as described in E180.

[0192] E182. The host cell as described in E181, wherein the host cell is a mammalian cell.

[0193] E183. The host cell as described in E182, wherein the host cell is a CHO cell, a HEK-293 cell, or a Sp2.0 cell.

[0194] E184. A kit comprising (i) the FAP antigen-binding protein as described in any one of E1-E178; the nucleic acid as described in E179; the vector as described in E180; the host cell as described in E182 or E183; or a combination thereof; and (ii) instructions for use.

[0195] E185. A pharmaceutical composition comprising (i) the FAP antigen-binding protein as described in any one of E1-E178; a nucleic acid as described in E179; a carrier as described in E180; a host cell as described in E182 or E183; or a combination thereof; and (ii) a pharmaceutically acceptable carrier, excipient, or diluent.

[0196] E185. A method for preparing a FAP antigen-binding protein as described in any one of E1-E178, the method comprising culturing host cells as described in any one of E181-E181 under conditions expressing the FAP antigen-binding protein.

[0197] E186. The method as described in E185, further comprising harvesting the expressed FAP antigen-binding protein.

[0198] E187. A method of treating cancer, the method comprising administering to a subject in need a therapeutically effective amount of FAP antigen-binding protein as described in any one of E1-E178 or a pharmaceutical composition as described in E185.

[0199] E188. As described in E187, wherein the cancer is a solid tumor.

[0200] E189. The method as described in E187 or E188, wherein the cancer comprises stromal cells.

[0201] E190. The method of any one of E187-E189, wherein the cancer comprises stromal cells expressing FAP.

[0202] E191. The method as described in any one of E187-190, wherein the subject is a human.

[0203] E192. The method as described in any one of E187-E191, wherein the FAP antigen-binding protein or the pharmaceutical composition is administered intravenously.

[0204] E193. The method as described in any one of E187-E191, wherein the FAP antigen-binding protein or the pharmaceutical composition is administered subcutaneously.

[0205] E194. The method of any one of E187-E193, wherein the FAP antigen-binding protein or the pharmaceutical composition is administered about twice a week, once a week, once every two weeks, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, once every eight weeks, once every nine weeks, once every ten weeks, twice a month, once a month, once every two months, once every three months, or once every four months.

[0206] E195. The FAP antigen-binding protein as described in any one of E1-E178 or the pharmaceutical composition as described in E185, for use as a medicine.

[0207] E196. The FAP antigen-binding protein as described in any one of E1-E178 or the pharmaceutical composition as described in E185, for use in treating a subject with cancer.

[0208] E197. Use of the FAP antigen-binding protein as described in any one of E1-E178 or the pharmaceutical composition as described in E185 in the manufacture of a medicament for treating a subject's cancer.

[0209] E198. Use of the FAP antigen-binding protein as described in any one of E1-E178 or the pharmaceutical composition as described in E185 for the treatment of a subject with cancer. Attached Figure Description

[0210] Figure 1A The crystal structure of FAP composite with scFv 32211 is shown. Figure 1B Epitopes and complementary residues and their interactions are shown.

[0211] Figure 2A The crystal structure of FAP composite with scFv 40828 is shown. Figure 2B Epitopes and complementary residues and their interactions are shown.

[0212] Figure 3A The crystal structure of the FAP composite with scFv 32229 is shown. Figure 3B Epitopes and complementary residues and their interactions are shown.

[0213] Figure 4A The crystal structure of FAP composite with scFv 40827 is shown. Figure 4B Epitopes and complementary residues and their interactions are shown.

[0214] Figure 5 This is a diagram of a bispecific molecule (IgG-scFv) containing an IgG moiety and two scFv moieties. A cysteine ​​clamp (represented by a horizontal line within the scFv) can be introduced into the scFv. The use (or absence) of a cysteine ​​clamp may require evaluation of the scFv's stability and biological activity. Detailed Implementation

[0215] 1. Overview Targeting T-cell costimulatory molecules has shown great promise in cancer therapy. However, agonists targeting these molecules also raise safety concerns, such as systemic toxicity (see, for example, Dahan et al., Cancer Cell 29, 820–831 (2016)). A preferred mechanism would be to limit the activation of T-cell costimulatory molecules within the cancer microenvironment.

[0216] FAP is an attractive target for delivering agonists of T-cell co-stimulatory molecules to cancerous tissue. FAP is highly expressed in the stroma of many solid tumors by cancer-associated fibroblasts (CAFs). FAP is selectively expressed in reactive stroma fibroblasts of over 90% of epithelial malignancies (primary and metastatic) (including lung, colorectal, bladder, ovarian, and breast cancer) and in malignant mesenchymal cells of bone and soft tissue sarcomas, whereas it is typically absent in normal adult tissues. FAP is also expressed on certain malignant tumor cells.

[0217] Therefore, the FAP antigen-binding protein disclosed in this paper can be used as a targeting component to deliver agonists of cancer therapeutic molecules, such as T-cell co-stimulatory molecules. T-cell co-stimulatory agonists can be engineered to be cross-link-dependent (also known as cluster-dependent), meaning that T-cell activation depends on the clustering of T-cell co-stimulatory molecules. In the absence of high levels of FAP (i.e., normal non-malignant cells), stimulation of T cells is minimal due to the lack of clustering, and immune activation and toxicity will be limited. Conversely, FAP is highly expressed in cancer-associated fibroblasts (CAFs); therefore, through FAP binding, a large number of T-cell agonists will approach each other, triggering receptor clustering and immune cell activation. This strategy has two major advantages: because activation will be primarily confined to tissues expressing FAP, systemic toxicity should be limited, and tumor-mediated T-cell activation leads to the killing of cancer cells.

[0218] The inventors also discovered that although FAP is a membrane-bound protein, its extracellular domain can detach and circulate in the peripheral blood. In fact, soluble FAP (sFAP) has been detected in both healthy individuals and diseased subjects (including cancer patients). If FAP antigen-binding proteins can bind to sFAP and form higher-order molecular complexes (i.e., multiple sFAP molecules complexed with multiple copies of FAP antigen-binding protein), systemic toxicity remains a risk due to the potential activation of immune cells in the peripheral blood. Therefore, in some cases, it is desirable to use an FAP antigen-binding protein that triggers cross-linking of T cell co-stimulatory molecules without binding to sFAP.

[0219] Therefore, this disclosure provides a FAP antigen-binding protein that targets cancer-associated fibroblasts. Such FAP antigen-binding molecules can be used to deliver cancer therapeutic molecules, such as agonists of T-cell co-stimulatory molecules, into cancer tissue, thereby limiting systemic toxicity.

[0220] 2. Antigen-binding proteins 2.1 Types of antigen-binding proteins The antigen-binding protein disclosed herein may take any of the many forms of antigen-binding proteins known in the art. In an exemplary aspect, the antigen-binding protein is an antibody or immunoglobulin, or an antigen-binding fragment of an antibody or immunoglobulin, or an antibody protein product.

[0221] In summary, antibodies belong to a family of plasma proteins called immunoglobulins and are composed of immunoglobulin domains (Janeway et al., *Immunobiology: The Immune System in Health and Disease*, 4th ed., Elsevier Science Ltd. / Garland Publishing, 1999). As used herein, the term "antibody" refers to a protein that has a conventional immunoglobulin form, contains both heavy and light chains, and includes variable and constant regions. For example, an antibody can be IgG, which is a "Y-shaped" structure of two pairs of identical polypeptide chains, each pair having a "light" chain (typically with a molecular weight of about 25 kDa) and a "heavy" chain (typically with a molecular weight of about 50–70 kDa). Antibodies have variable and constant regions. In the IgG form, the variable region is typically about 100–110 or more amino acids, contains three complementarity-determining regions (CDRs), is primarily responsible for antigen recognition, and is significantly different from other antibodies that bind to different antigens. The constant region allows antibodies to recruit cells and molecules of the immune system. The variable region consists of the N-terminal region of each light chain and heavy chain, while the constant region consists of the C-terminal portion of each heavy chain and light chain (Janeway et al., “Structure of the Antibody Molecule and the Immunoglobulin Genes”, Immunobiology: The Immune System in Health and Disease, 4th ed., Elsevier Science Ltd. / Garland Publishing, (1999)).

[0222] Antibodies may contain any constant region known in the art. Human light chains are classified as κ light chains and λ light chains. Heavy chains are classified as μ, δ, γ, α, or ε, and antibody isotypes are defined as IgM, IgD, IgG, IgA, and IgE, respectively. IgG has several subclasses, including but not limited to IgG1, IgG2, IgG3, and IgG4. IgM has subclasses, including but not limited to IgM1 and IgM2. The embodiments disclosed herein include all such antibody classes or isotypes. Light chain constant regions may be, for example, κ or λ type light chain constant regions, such as human κ or λ type light chain constant regions. Heavy chain constant regions may be, for example, α, δ, ε, γ, or μ type heavy chain constant regions, such as human α, δ, ε, γ, or μ type heavy chain constant regions. Therefore, in exemplary embodiments, the antibody is an antibody of isotype IgA, IgD, IgE, IgG, or IgM (including any one of IgG1, IgG2, IgG3, or IgG4).

[0223] Antibodies can be monoclonal or polyclonal. In some embodiments, an antibody comprises a sequence substantially similar to naturally occurring antibodies produced by mammals (e.g., mice, rabbits, goats, horses, chickens, hamsters, humans, etc.). In this respect, antibodies can be considered mammalian antibodies, such as mouse antibodies, rabbit antibodies, goat antibodies, horse antibodies, chicken antibodies, hamster antibodies, human antibodies, etc. In some aspects, antibodies are human antibodies. In some aspects, antibodies are chimeric antibodies or humanized antibodies. The term "chimeric antibody" refers to an antibody containing domains from two or more different antibodies. Chimeric antibodies may, for example, contain a constant domain from one species and a variable domain from a second species, or more generally, may contain segments of amino acid sequences from at least two species. Chimeric antibodies may also contain domains from two or more different antibodies within the same species. The term "humanized" in relation to the use of an antibody refers to an antibody having at least a non-human source CDR region engineered to have a structure and immunological function more similar to a real human antibody than the original source antibody. For example, humanization can involve transplanting a CDR from a non-human antibody (such as a mouse antibody) into a human antibody. Humanization can also involve selective amino acid substitutions to make the non-human sequence more similar to a human sequence.

[0224] Antibodies can be cleaved into fragments by enzymes such as papain and pepsin. Papain cleaves the antibody to produce two Fab fragments and a single Fc fragment. Pepsin cleaves the antibody to produce an F(ab')2 fragment and a pFc' fragment. In an exemplary aspect of this disclosure, the antigen-binding protein of this disclosure comprises an antigen-binding fragment of an antibody. As used herein, an "antigen-binding fragment" of an antibody refers to the portion of the antibody molecule that retains the ability to specifically bind to an antigen (preferably having substantially the same binding affinity). Examples of antigen-binding fragments include, but are not limited to: (i) Fab fragments, which are monovalent fragments consisting of VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, which are bivalent fragments consisting of two Fab fragments connected at the hinge region by disulfide bridges; (iii) Fd fragments consisting of VH and CH1 domains; (iv) Fv fragments consisting of VL and VH domains of an antibody arm; and (v) dAb fragments (Ward et al., 1989 Nature 341:544-546), which consist of VH domains.

[0225] Antibody architectures have been used to generate an increasing number of alternative forms that span a molecular weight range of at least about 12–150 kDa and have a range of valences (n) from monomers (n = 1), to dimers (n = 2), to trimers (n = 3), to tetramers (n = 4) and possibly higher; such alternative forms are referred to herein as “antibody protein products”.

[0226] The most commonly used structural unit for generating novel antibody-based forms is the single-chain variable (V) domain antibody fragment (scFv). Although the two domains, VL and VH, of the Fv fragment are encoded by separate genes, these two domains can be linked by a synthetic linker using recombination methods, which allows them to form a single protein chain in which the VL and VH regions pair to form a monovalent molecule (see, for example, Bird et al., Science [Science] 242:423-426 (1988) and Huston et al., 1988, Proc. Natl. Acad. Sci. USA [Proceedings of the National Academy of Sciences] 85:5879-5883).

[0227] It also covers other forms of single-chain antibody protein products, such as biantibodies. Biantibodies are bivalent, bispecific antibody protein products in which the VH and VL domains are expressed on a single polypeptide chain, but the linker used is too short to allow pairing between the two domains on the same chain, thus forcing the domains to pair with complementary domains on another chain and creating two antigen-binding sites (see, for example, Holliger et al., 1993, Proc. Natl. Acad. Sci. USA [Proceedings of the National Academy of Sciences] 90:6444-6448; Poljak et al., 1994, Structure [Structure] 2:1 121-1 123).

[0228] Other antibody protein products include disulfide-stabilized scFv (ds-scFv), single-chain Fab (scFab), and dimer and multimeric antibody forms (such as biantibodies, triantibodies, and tetraantibodies), or various forms of mini-abs consisting of scFvs linked to oligomeric domains. The smallest fragments are the VHH / VH of camel heavy chain Abs and single-domain Abs (sdAbs). Peptibody, or peptide-Fc fusion, is another type of antibody protein product. The structure of a peptibody consists of a bioactive peptide grafted onto an Fc domain. Peptibody has been well described in the art. See, for example, Shimamoto et al., mAbs [Monoclonal Antibodies] 4(5): 586-591 (2012).

[0229] Bispecific forms can generally be classified into five main categories: BsIgG, additional IgG, BsAb fragments, bispecific fusion proteins, and BsAb conjugates. See, for example, Spiess et al., Molecular Immunology 67(2) Part A: 97-106 (2015).

[0230] In an exemplary aspect, the antigen-binding protein disclosed herein comprises any of these antibody protein products. In an exemplary aspect, the antigen-binding protein disclosed herein comprises any of the following: scFv, Fab, Fv fragment, ds-scFv, scFab, dimer antibody, multimeric antibody (e.g., biantibody, triantibody, tetraantibody), miniAb, peptide, VHH / VH of camelid heavy chain antibody, sdAb, biantibody; triantibody; tetraantibody; bispecific or trispecific antibody, BsIgG, attached IgG, BsAb fragment, bispecific fusion protein, and BsAb conjugate.

[0231] In exemplary embodiments, the antigen-binding proteins disclosed herein comprise antibody protein products in monomeric or polymeric, oligomeric, or multimeric form. In some embodiments where the antibody comprises two or more distinct antigen-binding region fragments, the antibody is considered bispecific, trispecific, or multispecific, or divalent, trivalent, or multivalent, depending on the number of distinct epitopes recognized and bound by the antibody.

[0232] Many antigen-binding proteins disclosed in this article contain two distinct chains: a heavy chain derived from the antibody and a light chain derived from the antibody. Although the heavy / light chain has been modified and is no longer the classic immunoglobulin heavy / light chain, for convenience, it is often still referred to as the "heavy chain" or "HC" if it is based on the heavy chain backbone, and as the "light chain" or "LC" if it is based on the light chain backbone. For example, for the tetravalent bispecific molecule IgG-scFv, "HC" contains the IgG heavy chain fused to scFv. It is obvious to those skilled in the art whether HC represents a traditional immunoglobulin heavy chain or a modified form based on the immunoglobulin heavy chain backbone.

[0233] 2.2 Connection Position Some exemplary antigen-binding proteins disclosed herein are characterized by the epitopes they bind to or the complementary sites they contain. An "epitaph" refers to the range or region of an antigen that an antigen-binding protein specifically binds to, for example, the range or region containing residues that interact with the antigen-binding protein. Epitopes can be linear or conformational. Epitopes can be determined by any method well known in the art. For example, epitopes can be determined by routine immunoassays. Alternatively, antigen-binding proteins that bind to the same epitopes can be competitively screened. This is achieved by conducting competitive and cross-competitive studies to discover antigen-binding proteins that compete or cross-competitively bind to antigens such as FAP.

[0234] The term "complementary site" is derived from the definition of "epitope" above by reversing the perspective, and refers to the range or region in an antigen-binding protein that participates in antigen binding, for example, the range or region containing residues that interact with the antigen. Complementary sites can be linear or conformational (such as discontinuous residues in a CDR).

[0235] Epitopes / complementary sites can be defined and characterized at different levels of detail using a variety of experimental and computational epitope mapping methods. Experimental methods include mutagenesis, X-ray crystallography, nuclear magnetic resonance (NMR) spectroscopy, hydrogen / deuterium exchange mass spectrometry (HX-MS), cryo-electron microscopy, and various competitive binding methods. Because each method relies on unique principles, the description of an epitope is related to the method used to determine it. Therefore, the epitope / complementary site for a given binding pair will be defined differently depending on the mapping method employed.

[0236] At its most detailed level, epitopes / complementary sites in the interaction between antigen and antigen-binding protein can be defined by the spatial coordinates of the atomic contacts present in the interaction and information about their relative contributions to the binding thermodynamics. At one level, epitope / complementary site residues can be characterized by defining the spatial coordinates of the atomic contacts between the binding pairs. On one hand, epitope / complementary site residues can be defined by specific criteria, such as the distance between atoms in the antigen and atoms in the antigen-binding protein (e.g., a distance of 4.5 Å or less between the heavy atoms of the antigen and the heavy atoms of the antigen-binding protein (“contact” residues)). On the other hand, epitope / complementary site residues can be characterized as participating in hydrogen bonding interactions with homologous antibodies / antigens or with water molecules that also hydrogen bond with the antigen / antigen-binding protein (water-mediated hydrogen bonding). On yet another hand, epitope / complementary site residues can be characterized as forming salt bridges with residues of homologous antibodies / antigens. On yet another hand, epitope / complementary site residues can be characterized as residues with a non-zero buried surface area (BSA) variation due to the interaction between the antigen and antigen-binding protein.

[0237] At a less detailed level, epitopes / complementary sites can be characterized by function (e.g., through competitive binding with other antigen-binding molecules). Epitopes / complementary sites can also be more generally defined as containing amino acid residues to which substitution of another amino acid would alter the characteristics of the interaction between the binding pair (e.g., alanine scan).

[0238] In the context of X-ray derived crystal structures or cryo-electron microscopy structures, as illustrated herein with respect to FAP antigen-binding proteins, unless otherwise specified, FAP epitope residues refer to FAP residues that: (i) have a heavy atom (i.e., a non-hydrogen atom) within 4.5 Å of the heavy atom of the antigen-binding protein (also referred to as a "contact" residue); (ii) are involved in hydrogen bonding with a residue of the antigen-binding protein or with a water molecule that also hydrogen bonds with the antigen-binding protein (water-mediated hydrogen bonding); (iii) are involved in a salt bridge with a residue of the antigen-binding protein; and / or (iv) have a non-zero buried surface area (BSA) change due to interaction with the antigen-binding protein. Generally, a cutoff value is imposed on the BSA to avoid including residues with minimal interaction. Therefore, unless otherwise specified, if the epitope residue under category (iv) has a 20 Å... 2Or a larger BSA, or an epitope residue that participates in electrostatic interactions when the antigen-binding protein binds to the FAP, then the epitope residue is selected. Similarly, in the context of X-ray derived crystal structures or cryo-electron microscopy structures, unless otherwise stated or contradicted by the context, complementary site residues refer to antigen-binding protein residues that: (i) have a heavy atom (i.e., a non-hydrogen atom) within 4.5 Å of the heavy atom of the FAP (also referred to as “contact” residues), (ii) participate in hydrogen bonding with an antigen residue or with a water molecule that also hydrogen bonds with the FAP (water-mediated hydrogen bonding), (iii) participate in a salt bridge with a residue of the FAP, and / or (iv) have a non-zero buried surface area change due to interaction with the FAP. Likewise, unless otherwise stated, if the complementary site residue under category (iv) has 20 Å... 2 Alternatively, a larger BSA, or a complementary residue selected if the complementary residue participates in electrostatic interactions when the antigen-binding protein binds to the FAP.

[0239] Depending on the epitope mapping methods used and obtained at different levels of detail, it can be concluded that epitopes of different antigen-binding proteins on the same antigen can be compared similarly at different levels of detail. For example, epitopes described at the amino acid level (e.g., determined by X-ray or cryo-electron microscopy structures) contain the same set of amino acid residues, and these epitopes are considered identical. If epitopes do not share amino acid residues, these epitopes are considered independent (unique). If the binding of two molecules is mutually exclusive (i.e., the binding of one molecule excludes the simultaneous or successive binding of another molecule), epitopes characterized by competitive binding are considered overlapping; and if the antigen is adapted to the simultaneous binding of two molecules, these epitopes are considered independent (unique).

[0240] FAP exhibits prolyl endopeptidase activity and is active as a homodimer specific to type I collagen. For tumor targeting, it may be desirable for antigen-binding proteins not to bind to epitope residues located at or near the dimer interface. This is because "valence" may play a role in FAP-mediated receptor clustering. If the FAP conjugate binds at or near the dimer interface, only one copy of the FAP conjugate can bind to each FAP dimer. If the FAP conjugate (such as the exemplary FAP conjugate disclosed herein) binds to a region not near or at the dimer interface, both copies of the FAP conjugate can bind to each FAP dimer simultaneously. The simultaneous binding of two FAP conjugates to FAP dimers may enhance the ability of T cell co-stimulatory molecules to cluster (crosslink).

[0241] In addition to membrane-bound FAP, FAP can also detach from the membrane and exist in plasma in a soluble form. As discussed above, for tumor targeting, strong binding of soluble FAP is undesirable because binding to soluble FAP in plasma can activate circulating immune cells (e.g., B cells), thereby increasing the potential risk of systemic toxicity. The exemplary FAP-binding proteins disclosed herein are generally selected to demonstrate strong binding to membrane-bound FAP and weak binding to soluble FAP. In particular, molecules bound to group A (Table 5 in the examples) have been found to exhibit very low levels of soluble FAP binding. Preliminary observations based on negative staining electron microscopy indicate that systemic toxicity remains a risk if FAP antigen-binding proteins bind to sFAP and form molecular complexes (i.e., multiple sFAP molecules complexed with multiple copies of FAP antigen-binding proteins). Therefore, if the FAP antigen-binding protein has two arms (such as conventional Y-shaped immunoglobulins), the preferred epitope site should be located where neither arm can bind to the sFAP molecule simultaneously. Simultaneous binding of sFAP molecules on each arm may increase the risk of forming molecular complexes. For example, the location of epitope group A is believed to make it physically difficult to accommodate two FAPs (one on each arm).

[0242] In some embodiments, this disclosure provides a FAP antigen-binding protein that binds to an epitope comprising residues V275, R175, F181, Q182, I183, D178, F185, P179, and Y274. In some embodiments, the epitope may further comprise one or more residues selected from the group consisting of Q174, W327, P277, and Q278. In some embodiments, the epitope may further comprise one or more residues selected from the group consisting of P272, P180, I267, G276, D326, and A273. These epitope residues are numbered according to SEQ ID NO:394 (human FAP). Relevant residues from other FAP homologs, isotypes, variants, or fragments can be identified based on sequence alignment or structural alignment known in the art. For example, alignment can be performed manually or using a well-known sequence alignment program (such as ClustalW2 or “BLAST 2 sequence”) with default parameters. As illustrated by the structural data, V275, R175, F181, Q182, I183, D178, F185, P179, and Y274 were found to be “major” residues for scFv32211 binding; Q174, W327, P277, and Q278 were found to be “contributing” residues for scFv32211 binding; and P272, P180, I267, G276, D326, and A273, although “in contact” with residues of scFv32211, were still found to be “optional” residues for scFv32211 binding.

[0243] Based on structural studies, one or more of the following substitutions of the "major" residues may substantially disrupt the binding of scFv32211 to FAP: (1) R175 is replaced by A, N, D, C, Q, E, G, I, L, M, F, P, S, T, W, T, or V; (2) D178 is replaced by R, K, A, N, C, Q, G, I, L, M, F, P, S, T, W, Y, or V; (3) P179 is replaced by R, K, N, Q, I, M, F, W, Y, D, E, or H; (4) F181 is replaced by R, K, A, N, C, D, E, Q, G, I, L, M, P, S, T, V, or H; (5) F185 is replaced by R, K, A, N, C, D, E, Q, G, I, L, M, P, S, T, V, or H; (6) Y274 is replaced with R, K, A, G, I, L, M, P, S, T, V, H, F, or W; or (7) V275 is replaced with R, K, N, Q, I, L, M, H, F, W, Y, D, or E. One or more of the following substitutions of “minor” residues may substantially disrupt the binding of scFv32211 to FAP: (8) Q174 is replaced with A, C, G, I, L, M, F, P, W, Y, or V; (9) P277 is replaced with R, K, N, Q, I, L, M, H, F, W, Y, D, or E; or (10) Q278 is replaced with A, R, K, I, L, M, F, W, or Y. One or more of the following substitutions of the “minor” residues may substantially disrupt the binding of scFv32211 to FAP: (11) A273 is replaced by R, K, N, Q, I, M, F, W, Y, D, E or H; or (12) G276 is replaced by R, K, N, Q, I, L, M, H, F, W, Y, D or E.

[0244] In some embodiments, this disclosure provides a FAP antigen-binding protein that binds to an epitope comprising residues E325, D331, Q336, I320, R324, E302, R303, and T335. In some embodiments, the epitope may further comprise one or more residues selected from the group consisting of K381, D322, P333, and F357. In some embodiments, the epitope may further comprise one or more residues selected from the group consisting of K219, D326, W327, A361, I362, S363, Y364, I380, D382, F323, and H338. These epitope residues are numbered according to SEQ ID NO:394 (human FAP). Relevant residues from other FAP homologs, isotypes, variants, or fragments can be identified by sequence alignment or structural alignment known in the art. For example, alignment can be performed manually or using a well-known sequence alignment program (such as ClustalW2 or “BLAST 2 sequence”) with default parameters. As illustrated by the structural data, E325, D331, Q336, I320, R324, E302, R303, and T335 were found to be “major” residues for scFv40828 binding; K381, D322, P333, and F357 were found to be “contributing” residues for scFv40828 binding; and K219, D326, W327, A361, I362, S363, Y364, I380, D382, F323, and H338, although “contacting” residues of scFv40828, were still found to be “optional” residues for scFv40828 binding.

[0245] Based on structural studies, one or more of the following substitutions of the "major" residues may substantially disrupt the binding of scFv40828 to FAP: (1) E302 is replaced with A; (2) R303 is replaced with A; (3) I320 is replaced with A; (4) R324 is replaced with A; (5) E325 is replaced with A; (6) D331 is replaced with A; (7) T335 is replaced with A; or (8) Q336 is replaced with A. One or more of the following substitutions of the "minor" residues may substantially disrupt the binding of scFv40828 to FAP: (9) D322 is replaced with A; or (10) P333 is replaced with W, R, K, F, H, I, M, or Y; or F357 is replaced with A. One or more of the following substitutions of the “minor” residues may substantially disrupt the binding of scFv40828 to FAP: (11) Y364 is replaced with A; or (12) D382 is replaced with W, R, K, F, H, I, M or Y.

[0246] In some embodiments, this disclosure provides a FAP antigen-binding protein that binds to an epitope comprising residues S86, E82, I181, T83, Q85, Q65, K486, N80, T88, I485, and I487. In some embodiments, the epitope may further comprise one or more residues selected from the group consisting of V77, G84, Y79, and Y87. In some embodiments, the epitope may further comprise one or more residues selected from the group consisting of E66, Y67, S71, D73, N75, S136, and L488. These epitope residues are numbered according to SEQ ID NO:394 (human FAP). Relevant residues from other FAP homologs, isotypes, variants, or fragments can be identified based on sequence alignment or structural alignment known in the art. For example, alignment can be performed manually or using a well-known sequence alignment program (such as ClustalW2 or "BLAST2 sequence") with default parameters. As illustrated by the structural data, S86, E82, I181, T83, Q85, Q65, K486, N80, T88, I485, and I487 were found to be “major” residues for scFv32229 binding; V77, G84, Y79, and Y87 were found to be “contributing” residues for scFv32229 binding; and E66, Y67, S71, D73, N75, S136, and L488, although “in contact” with residues of scFv32229, were still found to be “optional” residues for scFv32229 binding.

[0247] Based on structural studies, one or more of the following substitutions of the "major" residues may substantially disrupt the binding of scFv32229 to FAP: (1) Q65 is replaced with A; (2) N80 is replaced with A; (3) E82 is replaced with A; (4) T83 is replaced with A; (5) Q85 is replaced with A; (6) S86 is replaced with A; (7) T88 is replaced with A; (8) I485 is replaced with A; or (9) K486 is replaced with A. One or more of the following substitutions of the "minor" residues may substantially disrupt the binding of scFv32229 to FAP: (10) V77 is replaced with A; (11) Y79 is replaced with A; (12) G84 is replaced with A; or (13) Y87 is replaced with A. One or more of the following substitutions of “minor” residues may substantially disrupt the binding of scFv32229 to FAP: (14) N75 is replaced with F, Y or W.

[0248] In some embodiments, this disclosure provides a FAP antigen-binding protein that binds to an epitope comprising residues K381, K371, E414, S428, Q389, Y432, I390, P434, and K436. In some embodiments, the epitope may further comprise one or more residues selected from the group consisting of I427, I388, G430, P433, and I426. In some embodiments, the epitope may further comprise one or more residues selected from the group consisting of W395, S435, Y379, N386, A387, and S392. These epitope residues are numbered according to SEQ ID NO:394 (human FAP). Relevant residues from other FAP homologs, isotypes, variants, or fragments can be identified using sequence alignment or structural alignment known in the art. For example, alignment can be performed manually or using a well-known sequence alignment program (such as ClustalW2 or “BLAST 2 sequence”) with default parameters. As illustrated by the structural data, K381, K371, E414, S428, Q389, Y432, I390, P434, and K436 were found to be “major” residues for scFv40827 binding; I427, I388, G430, P433, and I426 were found to be “contributing” residues for scFv40827 binding; and W395, S435, Y379, N386, A387, and S392, although “in contact” with residues of scFv40827, were still found to be “optional” residues for scFv40827 binding.

[0249] Based on structural studies, one or more of the following substitutions of the "major" residues may substantially disrupt the binding of scFv40327 to FAP: (1) K371 is replaced with A; (2) K381 is replaced with A; (3) E414 is replaced with A; (4) I390 is replaced with A; (5) Y432 is replaced with A; (6) P434 is replaced with F; or (7) K436 is replaced with A. One or more of the following substitutions of the "minor" residues may substantially disrupt the binding of scFv40827 to FAP: (8) I388 is replaced with A; (9) I427 is replaced with A; (10) G430 is replaced with A; or (11) P433 is replaced with F.

[0250] Disruption of binding can be assessed by measuring the affinity of the antigen-binding molecule for the epitope (e.g., KD value); or by competitive assays known in the art.

[0251] 2.3 Structure of FAP antigen-binding protein In some embodiments, this disclosure provides an FAP antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the FAP antigen-binding protein comprises the following complementary residues (VH and VL numbered according to Kabat): (1) H33 is Arg, Lys, Gln, or Asn; (2) H94 is Arg, Lys, Gln, or Asn; (3) H97 is Gly or Ala; (4) H98 is Tyr, Trp, Phe, Thr, or Ser; (5) H100B is Tyr, Trp, Phe, Thr, or Ser; (6) H100C is Tyr, Trp, Phe, Thr, or Ser; (7) L53 is Gln, Asn, or Glu; (8) L54 is Arg, Lys, Gln, or Asn; and (9) L60 is Asp, Glu, or Asn. Based on structural studies, these complementary residues were found to be the “major” complementary residues from the FAP-binding clone scFv32211. Preferably, (1) H33 is Arg or Lys; (2) H94 is Arg or Lys; (3) H97 is Gly or Ala; (4) H98 is Tyr or Phe; (5) H100B is Tyr or Phe; (6) H100C is Tyr or Phe; (7) L53 is Gln or Asn; (8) L54 is Arg or Lys; and (9) L60 is Asp or Glu. More preferably, (1) H33 is Arg; (2) H94 is Arg; (3) H97 is Gly; (4) H98 is Tyr; (5) H100B is Tyr; (6) H100C is Tyr; (7) L53 is Gln; (8) L54 is Arg; and (9) L60 is Asp.

[0252] The FAP antigen-binding protein may further contain the following complementary residues (VH and VL numbered according to Kabat): (10) H31 is Asn, Gln, His, Asp, Lys, or Arg; (11) H34 is Val, Ile, Leu, Met, Phe, Ala, or leucine; (12) L49 is Tyr, Trp, Phe, Thr, or Ser; and (13) L50 is Ser or Thr. Based on structural studies, these complementary residues have been found to be “contributing” residues from the FAP-binding clone scFv32211. Preferably, (10) H31 is Asn or Gln; (11) H34 is Val or Leu; (12) L49 is Tyr or Phe; and (13) L50 is Ser or Thr. More preferably, (10) H31 is Asn; (11) H34 is Val; (12) L49 is Tyr; and (13) L50 is Ser.

[0253] The FAP antigen-binding protein may further contain the following complementary residues (VH and VL numbered according to Kabat): (14) H27 is Phe, Leu, Val, Ile, Ala, or Tyr; (15) H28 is Ser or Thr; (16) H30 is Ser or Thr; (17) H95 is Ile, Leu, Val, Met, Ala, Phe, or leucine; (18) H96 is Gly or Ala; (19) H101 is Asp, Glu, or Asn; and (20) L52 is Asn, Gln, His, Asp, Lys, or Arg. Based on structural studies, these complementary residues were found to be “optional” residues from the FAP-binding clone scFv32211. Preferably, (14) H27 is Phe or Tyr; (15) H28 is Ser or Thr; (16) H30 is Ser or Thr; (17) H95 is Ile or Leu; (18) H96 is Gly or Ala; (19) H101 is Asp or Glu; and (20) L52 is Asn or Gln. More preferably, (14) H27 is Phe; (15) H28 is Ser; (16) H30 is Ser; (17) H95 is Ile; (18) H96 is Gly; (19) H101 is Asp; and (20) L52 is Asn.

[0254] Based on structural studies, it was found that for scFv32211, among the six complementarity-determining regions (CDRs), CDR-H1, CDR-H3, and CDR-L2 are responsible for contacting FAP residues. Furthermore, the VL framework residue L60 (kabat number) also forms a salt bridge with the FAP residue R175. Therefore, in some embodiments, the FAP antigen-binding protein comprises (a) VH, which includes: (i) a complementarity-determining region (CDR)-H1 containing any one of SEQ ID NO: 398-402; and (ii) a CDR-H3 containing any one of SEQ ID NO: 408-412; (b) VL, which includes a CDR-L2 containing any one of SEQ ID NO: 418-422; and (c) said VL further includes a D at position L60. In some embodiments, the CDR-H1, the CDR-H3, the CDR-L2, and the residue D at L60 are selected from one or more FAP residues of the group consisting of: Q174, R175, D178, P179, P180, F181, Q182, I183, F185, I267, P272, A273, Y274, V275, G276, P277, Q278, D326, and W327 (according to SEQ ID NO:394).

[0255] In some embodiments, this disclosure provides an FAP antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the FAP antigen-binding protein comprises the following complementary residues (VH and VL numbered according to Kabat): (1) H31 is Arg, Lys, Gln, or Asn; (2) H98 is Tyr, Trp, Phe, Thr, or Ser; (3) H99 is Tyr, Trp, Phe, Thr, or Ser; (4) H100 is Tyr, Trp, Phe, Thr, or Ser; and (5) H101 is Asp, Glu, or Asn. Based on structural studies, these complementary residues were found to be “major” complementary residues from the FAP-binding clone scFv40828. Preferably, (1) H31 is Arg or Lys; (2) H98 is Tyr or Phe; (3) H99 is Tyr or Phe; (4) H100 is Tyr or Phe; and (5) H101 is Asp or Glu. More preferably, (1) H31 is Arg; (2) H98 is Tyr; (3) H99 is Tyr; (4) H100 is Tyr; and (5) H101 is Asp.

[0256] The FAP antigen-binding protein may further include the following complementary residues (VH and VL numbered according to Kabat): (6) H28 is Thr or Ser; and (7) H100A is Tyr, Trp, Phe, Thr, or Ser. Based on structural studies, these complementary residues have been found to be “contributing” residues from the FAP-binding clone scFv40828. Preferably, (6) H28 is Thr or Ser; and (7) H100A is Tyr or Phe. More preferably, (6) H28 is Thr; and (7) H100A is Tyr.

[0257] The FAP antigen-binding protein may further contain the following complementary residues (VH and VL numbered according to Kabat): (8) H32 is Tyr, Trp, Phe, Thr, or Ser; (9) H96 is Pro or Ala; (10) H97 is Ser or Thr; (11) H100C is Tyr, Trp, Phe, Thr, or Ser; (12) L32 is Phe, Leu, Val, Ile, Ala, or Tyr; and (13) L49 is Tyr, Trp, Phe, Thr, or Ser. Based on structural studies, these complementary residues were found to be “optional” residues from the FAP-binding clone scFv40828. Preferably, (8) H32 is Tyr or Phe; (9) H96 is Pro or Ala; (10) H97 is Ser or Thr; (11) H100C is Tyr or Phe; (12) L32 is Phe or Tyr; and (13) L49 is Tyr or Phe. More preferably, (8) H32 is Tyr; (9) H96 is Pro; (10) H97 is Ser; (11) H100C is Tyr; (12) L32 is Phe; and (13) L49 is Tyr.

[0258] Based on structural studies, it was found that for scFv40828, among the six complementarity-determining regions (CDRs), CDR-H1, CDR-H3, and CDR-L2 are responsible for contacting FAP residues. Therefore, in some embodiments, the FAP antigen-binding protein comprises (a) VH, which comprises: (i) a complementarity-determining region (CDR)-H1 containing any one of SEQ ID NO: 428-432; and (ii) a CDR-H3 containing any one of SEQ ID NO: 438-442; and (b) VL, which comprises a CDR-L2 containing any one of SEQ ID NO: 448-452. In some embodiments, CDR-H1, CDR-H3, and CDR-L2 contact one or more FAP residues selected from the group consisting of: K219, E302, R303, I320, D322, F323, R324, E325, D326, W327, D331, P333, T335, Q336, H338, F357, A361, I362, S363, Y364, I380, K381, and D382 (according to SEQ ID NO: 394).

[0259] In some embodiments, this disclosure provides an FAP antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the FAP antigen-binding protein comprises the following complementary residues (VH and VL numbered according to Kabat): (1) H31 is Asn, Gln, His, Asp, Lys, or Arg; (2) H33 is Gly or Ala; (3) H52A is Tyr, Trp, Phe, Thr, or Ser; (4) H55 is Arg, Lys, Gln, or Asn; (5) H56 is Asn, Gln, His, Asp, Lys, or Arg; (6) H95 is Asp, Glu, or Asn; (7) H100 is Gly or Ala; (8) L32 is Tyr, Trp, Phe, Thr, or Ser; (9) L91 is Phe, Leu, Val, Ile, Ala, or Tyr; and (10) L95 is Tyr, Trp, Phe, Thr, or Ser. Based on structural studies, these complementary residues were found to be “major” complementary residues from the FAP-binding clone scFv32229. Preferably, (1) H31 is Asn or Gln; (2) H33 is Gly or Ala; (3) H52A is Tyr or Phe; (4) H55 is Arg or Lys; (5) H56 is Asn or Gln; (6) H95 is Asp or Glu; (7) H100 is Gly or Ala; (8) L32 is Tyr or Phe; (9) L91 is Phe or Tyr; and (10) L95 is Tyr or Phe. More preferably, (1) H31 is Asn; (2) H33 is Gly; (3) H52A is Tyr; (4) H55 is Arg; (5) H56 is Asn; (6) H95 is Asp; (7) H100 is Gly; (8) L32 is Tyr; (9) L91 is Phe; and (10) L95 is Tyr.

[0260] The FAP antigen-binding protein may further contain the following complementary residues (VH and VL numbered according to Kabat): (11) H28 is Thr or Ser; (12) H32 is Tyr, Trp, Phe, Thr or Ser; (13) H52 is Trp, Tyr or Phe; (14) L30 is Tyr, Trp, Phe, Thr or Ser; and (15) L96 is Trp, Tyr or Phe. Based on structural studies, these complementary residues were found to be “contributing” residues from the FAP-binding clone scFv32229. Preferably, (11) H28 is Thr or Ser; (12) H32 is Tyr or Phe; (13) H52 is Trp or Tyr; (14) L30 is Tyr or Phe; and (15) L96 is Trp or Tyr. More preferably, (11) H28 is Thr; (12) H32 is Tyr; (13) H52 is Trp; (14) L30 is Tyr; and (15) L96 is Trp.

[0261] The FAP antigen-binding protein may further contain the following complementary residues (VH and VL numbered according to Kabat): (16) H30 is Asn, Gln, His, Asp, Lys, or Arg; (17) H53 is Asp, Glu, or Asn; (18) H96 is Gly or Ala; (19) H97 is Ser or Thr; (20) H98 is Gly or Ala; (21) H99 is Gly or Ala; and (22) L56 is Ser or Thr. Based on structural studies, these complementary residues were found to be “optional” residues from the FAP-binding clone scFv32229. Preferably, (16) H30 is Asn or Gln; (17) H53 is Asp or Glu; (18) H96 is Gly or Ala; (19) H97 is Ser or Thr; (20) H98 is Gly or Ala; (21) H99 is Gly or Ala; and (22) L56 is Ser or Thr. More preferably, (16) H30 is Asn; (17) H53 is Asp; (18) H96 is Gly; (19) H97 is Ser; (20) H98 is Gly; (21) H99 is Gly; and (22) L56 is Ser.

[0262] Based on structural studies, it was found that for scFv32229, CDR-H1, CDR-H2, CDR-H3 and CDR-L2 contain residues that contact FAP residues in all six complementarity-determining regions (CDRs). Therefore, in some embodiments, the FAP antigen-binding protein comprises (a) VH, which comprises: (i) CDR-H1 comprising any one of SEQ ID NO: 458-462; (ii) CDR-H3 comprising any one of SEQ ID NO: 463-467; and (iii) CDR-H3 comprising any one of SEQ ID NO: 468-472; and (b) VL, which comprises: (i) CDR-L1 comprising any one of SEQ ID NO: 473-487; (ii) CDR-L2 comprising any one of SEQ ID NO: 478-482; and (iii) CDR-L3 comprising any one of SEQ ID NO: 483-487. In some embodiments, CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 contact one or more FAP residues selected from the group consisting of: Q65, N75, V77, Y79, N80, E82, T83, G84, Q85, S86, Y87, T88, I485, and K486 (according to SEQ ID NO:394).

[0263] In some embodiments, this disclosure provides an FAP antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the FAP antigen-binding protein comprises the following complementary residues (VH and VL numbered according to Kabat): (1) H28 is Thr or Ser; (2) H53 is Asp, Glu, or Asn; (3) H96 is Arg, Lys, Gln, or Asn; (4) H99 is Tyr, Trp, Phe, Thr, or Ser; (5) H100A is Tyr, Trp, Phe, Thr, or Ser; (6) H100B is Tyr, Trp, Phe, Thr, or Ser; and (7) H100C is Tyr, Trp, Phe, Thr, or Ser. Based on structural studies, these complementary residues were found to be “major” complementary residues from the FAP-binding clone scFv40827. Preferably, (1) H28 is Thr or Ser; (2) H53 is Asp or Glu; (3) H96 is Arg or Lys; (4) H99 is Tyr or Phe; (5) H100A is Tyr or Phe; (6) H100B is Tyr or Phe; and (7) H100C is Tyr or Phe. More preferably, (1) H28 is Thr; (2) H53 is Asp; (3) H96 is Arg; (4) H99 is Tyr; (5) H100A is Tyr; (6) H100B is Tyr; and (7) H100C is Tyr.

[0264] The FAP antigen-binding protein may further contain the following complementary residues (VH and VL numbered according to Kabat): (8) H31 is Ser or Thr; (9) H52 is Trp, Tyr, or Phe; (10) H97 is Leu, leucine, Ile, Val, Met, Ala, or Phe; (11) H98 is Gln, Asn, or Glu; (12) L32 is Leu, leucine, Ile, Val, Met, Ala, or Phe; (13) L91 is Tyr, Trp, Phe, Thr, or Ser; and (14) L96 is Trp, Tyr, or Phe. Based on structural studies, these complementary residues were found to be “contributing” residues from the FAP-binding clone scFv40827. Preferably, (8) H31 is Ser or Thr; (9) H52 is Trp or Tyr; (10) H97 is Leu or Ile; (11) H98 is Gln or Asn; (12) L32 is Leu or Ile; (13) L91 is Tyr or Phe; and (14) L96 is Trp or Tyr. More preferably, (8) H31 is Ser; (9) H52 is Trp; (10) H97 is Leu; (11) H98 is Gln; (12) L32 is Leu; (13) L91 is Tyr; and (14) L96 is Trp.

[0265] The FAP antigen-binding protein may further include the following complementary residues (VH and VL numbered according to Kabat): (15) H30 is Ser or Thr; (16) H95 is Asp, Glu, or Asn; (17) H100 is Asp, Glu, or Asn; and (18) L30 is Tyr, Trp, Phe, Thr, or Ser. Based on structural studies, these complementary residues were found to be “optional” residues from the FAP-binding clone scFv40827. Preferably, (15) H30 is Ser or Thr; (16) H95 is Asp and Glu; (17) H100 is Asp or Glu; and (18) L30 is Tyr and Phe. More preferably, (15) H30 is Ser; (16) H95 is Asp; (17) H100 is Asp; and (18) L30 is Tyr.

[0266] Based on structural studies, it was found that for scFv32229, CDR-H1, CDR-H2, CDR-H3, CDR-L1, and CDR-L3 contain residues that contact FAP residues in all six complementarity-determining regions (CDRs). Therefore, in some embodiments, the FAP antigen-binding protein comprises (a) VH, which comprises: (i) CDR-H1 comprising any one of SEQ ID NO: 488-492; (ii) CDR-H2 comprising any one of SEQ ID NO: 493-497; and (iii) CDR-H3 comprising any one of SEQ ID NO: 498-502; and (b) VL, which comprises: (i) CDR-L1 comprising any one of SEQ ID NO: 503-507; and (ii) CDR-L3 comprising any one of SEQ ID NO: 513-517. In some embodiments, CDR-H1, CDR-H2, CDR-H3, CDR-L1, and CDR-L3 are contacted with one or more FAP residues selected from the group consisting of: K381, K371, E414, S428, Q389, Y432, I390, P434, K436, I427, I388, G430, P433, I429, W395, S435, Y379, N386, A387, and S392 (according to SEQ ID NO:394).

[0267] As used herein, complementarity-determining regions (CDRs) can be identified based on the cumulative, AbM, contact, North, and / or conformational definitions of Kabat, Chothia, and both Kabat and Chothia, or any CDR identification method well known in the art. See, for example, Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th edition (hypervariable regions); Chothia et al., 1989, Nature 342:877-883 (structural loop structures). The identity of the amino acid residues constituting the CDR in a particular antibody can be determined using methods well known in the art. The AbM definition of the CDR is a compromise between Kabat and Chothia, and Oxford Molecular Diagnostics' Accelrys® AbM antibody modeling software was used. The “contact” definition of a CDR is based on observed antigen contact and is described in MacCallum et al., 1996, J. Mol. Biol. [Journal of Molecular Biology], 262:732-745. The “conformation” definition of a CDR is based on residues that contribute enthalpy to antigen binding (see, for example, Makabe et al., 2008, J. Biol. Chem. [Journal of Biochemistry], 283:1156-1166). North has identified the classical CDR conformation using a different set of preferred CDR definitions (North et al., 2011, J. Mol. Biol. [Journal of Molecular Biology] 406:228-256). In another approach referred to herein as the “conformation definition” of a CDR, the position of the CDR can be identified as residues that contribute enthalpy to antigen binding (Makabe et al., 2008, J Biol. Chem. [Journal of Biochemistry] 283:1156-1166).The Martin definition (also known as the enhanced Chothia definition) combines the Kabat and Chothia definitions and differs from them only on the heavy chain, where CDR-H1 includes all residues of both Kabat and Chothia, while CDR-H2 is seven residues shorter than the CDR-H2 defined by Kabat (Martin, Bioinformatics tools for antibody engineering. Handbook of Therapeutic Antibodies. Weinheim: Wiley-VCH Verlag GmbH; (2008). pp. 95–117; see also the database maintained by the School of Structural and Molecular Biology, University of London, http: / / www.bioinf.org.uk / abs / #cdrid). Other CDR boundary definitions may not strictly follow one of the above methods but will still overlap with at least a portion of the Kabat CDR; however, they can be shortened or lengthened based on predictions or experimental findings that specific residues or groups of residues or even the entire CDR do not significantly affect antigen binding. For example, a “combined” CDR may also be used. Therefore, a CDR can refer to a CDR defined by any method known in the art, including combinations of methods. For any given embodiment containing more than one CDR, the CDR (or other residues of the antibody) may be defined according to any of the following: Kabat, Chothia, North, AbM, Contact, IMGT, Martin (combined Kabat and Chothia), and / or conformational definitions.

[0268] For example, the table below shows several commonly used definitions of CDR: 1. Some of these definitions (particularly for the Chothia ring) vary depending on the individual publication examined. Some papers describe the Chothia CDR as follows: CDR-L1: L24-34; CDR-L2: L50-56; CDR-L3: L89-97; CDR-H1: H26-32; CDR-H2: H52-56; CDR-H3: H95-102.

[0269] 2. Except for the contact definition which uses the Chothia or Martin (enhanced Chothia) definition, any numbering scheme can be used for these CDR definitions.

[0270] 3. When using the Kabat numbering convention, the end of the Chothia CDR-H1 ring varies between H32 and H34 depending on the length of the ring (this is because the Kabat numbering scheme places the insertion at H35A and H35B).

[0271] The CDR sequences provided in the sequence listing are based on the Kabat definition. However, other definitions of CDR may also be used. Therefore, in some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) SEQ ID NO: 197, 199, 201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, Heavy chains CDR-H1, CDR-H2, and CDR-H3 of 277, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 332, 334, 335, 339, or 341; and (ii) SEQ ID NO: 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 2 Light chain CDR-L1, CDR-L2, and CDR-L3 of sequence numbers 76, 278, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 333, 336, 337, 338, 340, or 342. In some embodiments, as shown in sequence listing B, the three heavy chain CDRs and the three light chain CDRs are from the same clone. In exemplary embodiments, the CDRs are defined according to Kabat, Chothia, AbM, contact, or IMGT.

[0272] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:197; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:198. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0273] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:199; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:200. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0274] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:201; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:202. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0275] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:203; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:204. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0276] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:205; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:206. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0277] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:207; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:208. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0278] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:209; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:210. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0279] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:211; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:212. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0280] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:213; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:214. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0281] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:215; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:216. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0282] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:217; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:218. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0283] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:219; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:220. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0284] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:221; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:222. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0285] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:223; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:224. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0286] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:225; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:226. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0287] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:227; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:228. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0288] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:229; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:230. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0289] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:231; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:232. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0290] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:233; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:234. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0291] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:235; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:236. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0292] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:237; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:238. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0293] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:239; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:240. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0294] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:241; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:242. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0295] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:243; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:244. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0296] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:245; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:246. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0297] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:247; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:248. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0298] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:249; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:250. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0299] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:251; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:252. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0300] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:253; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:254. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0301] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:255; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:256. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0302] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:257; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:258. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0303] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:259; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:260. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0304] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:261; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:262. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0305] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:263; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:264. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0306] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:265; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:266. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0307] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:267; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:268. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0308] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:269; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:270. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0309] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:271; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:272. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0310] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:273; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:274. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0311] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:275; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:276. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0312] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:277; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:278. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0313] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:279; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:280. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0314] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:281; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:282. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0315] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:283; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:284. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0316] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:285; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:286. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0317] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:287; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:288. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0318] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:289; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:290. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0319] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:291; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:292. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0320] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:293; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:284. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0321] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:295; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:296. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0322] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:297; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:298. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0323] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:299; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:300. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0324] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:301; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:302. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0325] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:303; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:304. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0326] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:305; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:306. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0327] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:307; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:308. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0328] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:309; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:310. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0329] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:311; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:312. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0330] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:313; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:314. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0331] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:315; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:316. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0332] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:317; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:318. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0333] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:319; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:320. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0334] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:321; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:322. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0335] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:323; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:324. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0336] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:325; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:326. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0337] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:327; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:328. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0338] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:329; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:330. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0339] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:331; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:332. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0340] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:333; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:334. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0341] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:335; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:336. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or 50 nM or less.

[0342] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:337; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:338. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0343] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:339; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:340. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0344] In various embodiments, the FAP antigen-binding protein comprises a VH containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three heavy chain CDRs in SEQ ID NO:341; and a VL containing three CDRs, which, when combined, are at least 85%, at least 90%, or at least 95% identical to the three light chain CDRs in SEQ ID NO:342. The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0345] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 197, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 198.

[0346] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 199, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 200.

[0347] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 201, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 202.

[0348] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 203, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 204.

[0349] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 205, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 206.

[0350] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 207, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 208.

[0351] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 209, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 210.

[0352] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 211, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 212.

[0353] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 213, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 214.

[0354] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 215, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 216.

[0355] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 217, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 218.

[0356] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 219, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 220.

[0357] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 221, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 222.

[0358] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 223, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 224.

[0359] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 225, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 226.

[0360] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 227, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 228.

[0361] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 229, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 230.

[0362] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 231, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 232.

[0363] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 233, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 234.

[0364] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 235, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 236.

[0365] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 237, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 238.

[0366] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 239, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 240.

[0367] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 241, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 242.

[0368] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 243, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 244.

[0369] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 245, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 246.

[0370] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 247, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 248.

[0371] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 249, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 250.

[0372] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 251, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 252.

[0373] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 253, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 254.

[0374] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 255, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 256.

[0375] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 257, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 258.

[0376] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 259, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 260.

[0377] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 261, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 262.

[0378] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 263, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 264.

[0379] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 265, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 266.

[0380] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 267, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 268.

[0381] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 269, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 270.

[0382] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 271, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 272.

[0383] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 273, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 274.

[0384] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 275, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 276.

[0385] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 277, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 278.

[0386] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 279, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 280.

[0387] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 281, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 282.

[0388] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 283, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 284.

[0389] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 285, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 286.

[0390] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 287, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 288.

[0391] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 289, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 290.

[0392] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 291, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 292.

[0393] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 293, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 294.

[0394] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 295, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 296.

[0395] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 297, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 298.

[0396] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 299, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 300.

[0397] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 301, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 302.

[0398] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 303, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 304.

[0399] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 305, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 306.

[0400] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 307, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 308.

[0401] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 309, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 310.

[0402] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 311, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 312.

[0403] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 313, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 314.

[0404] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 315, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 316.

[0405] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 317, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 318.

[0406] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 319, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 320.

[0407] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 321, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 322.

[0408] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 323, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 324.

[0409] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 325, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 326.

[0410] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 327, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 328.

[0411] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 329, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 330.

[0412] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 331, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 332.

[0413] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 333, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 334.

[0414] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 335, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 336.

[0415] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 337, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 338.

[0416] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 339, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 340.

[0417] In some embodiments, the FAP antigen-binding protein disclosed herein comprises: (i) the heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 341, and (ii) the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 342.

[0418] In an exemplary aspect, the FAP antigen-binding protein comprises (a) the CDR-H1 amino acid sequence set forth in Sequence Listing A, Tables 10F, 11F, 12F, and 13F, or a variant sequence thereof differing only in 1-4 amino acids (e.g., 1, 2, 3, or 4 amino acids) or having at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence identity; (b) the CDR-H2 amino acid sequence set forth in Sequence Listing A, Tables 10F, 11F, 12F, and 13F, or a variant sequence thereof differing only in 1-4 amino acids (e.g., 1, 2, 3, or 4 amino acids) or having at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence identity; (c) (d) The CDR-H3 amino acid sequence described in Sequence Listing A, Tables 10F, 11F, 12F, and 13F, or a variant sequence thereof that differs only in 1-4 amino acids (e.g., 1, 2, 3, or 4 amino acids) or has at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence identity; (e) The CDR-L1 amino acid sequence described in Sequence Listing A, Tables 10F, 11F, 12F, and 13F, or a variant sequence thereof that differs only in 1-4 amino acids (e.g., 1, 2, 3, or 4 amino acids) or has at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence identity; The CDR-L2 amino acid sequence described in Sequence Listing A, Tables 10F, 11F, 12F, and 13F, or a variant sequence thereof differing only in 1-4 amino acids (e.g., 1, 2, 3, or 4 amino acids) or having at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence identity; (f) the CDR-L3 amino acid sequence described in Sequence Listing A, Tables 10F, 11F, 12F, and 13F, or a variant sequence thereof differing only in 1-4 amino acids (e.g., 1, 2, 3, or 4 amino acids) or having at least about 80%, at least about 85%, at least about 90%, or at least about 95% sequence identity; or (g) any combination of two, three, four, five, or six from (a) to (f). The antigen-binding protein may further bind to its target (FAP) with a KD value of 100 nM or less, or a KD value of 50 nM or less.

[0419] In an exemplary embodiment, the FAP antigen-binding protein comprises 3, 4, 5, or all 6 amino acid sequences identified by SEQ ID NO under the same clone name in Sequence Listings A, 10F, 11F, 12F, and 13F. In an exemplary embodiment, the antigen-binding protein comprises each of the LC CDR amino acid sequences identified by SEQ ID NO under the same clone name in Sequence Listings A, 10F, 11F, 12F, and 13F, and at least 1 or 2 of the HC CDR amino acid sequences identified by SEQ ID NO under the same clone name in Sequence Listings A, 10F, 11F, 12F, and 13F. In an exemplary embodiment, the FAP antigen-binding protein comprises each of the HCCDR amino acid sequences identified by SEQ ID NO under the same clone name in Sequence Listings A, 10F, 11F, 12F, and 13F, and at least 1 or 2 of the LC CDR amino acid sequences identified by SEQ ID NO under the same clone name in Sequence Listings A, 10F, 11F, 12F, and 13F.In an exemplary embodiment, the antigen-binding protein comprises six CDR amino acid sequences listed under the same clone names in Sequence Listing A, Tables 10F, 11F, 12F, and 13F, or comprises six CDR amino acid sequences selected from the group consisting of: (1) SEQ ID NO: 1-6, (2) SEQ ID NO: 7-12, (3) SEQ ID NO: 13-18, (4) SEQ ID NO: 19-24, (5) SEQ ID NO: 25-30, (6) SEQ ID NO: 31-36, (7) SEQ ID NO: 37-42, (8) SEQ ID NO: 43-48, (9) SEQ ID NO: 49-54, (10) SEQ ID NO: 55-60, (11) SEQ ID NO: 61-66, (12) SEQ ID NO: 67-72, (13) SEQ ID NO: 73-78, (14) SEQ ID NO: 79-84, (15) SEQ ID NO: 85-90, (16) SEQ ID NO: 91-96, (17) SEQ ID NO: 97-102, (18) SEQ ID NO: 103-108, (19) SEQ ID NO: 109-114, (20) SEQ ID NO: 115-120, (21) SEQ ID NO: 121-126, (22) SEQ ID NO: 127-132, (23) SEQ ID NO: 133-138, (24) SEQ ID NO: 139-144, (25) SEQ ID NO: 145-150, (26) SEQ ID NO: 151-156, (27) SEQ ID NO: 157-162, (28) SEQ ID NO: 163-168, (29) SEQ ID NO: 169-174, (30) SEQ ID NO: 175-180, (31) SEQ ID NO: 181-186, (32) SEQ ID NO: 187-192, (33) SEQ ID NO: 43-47 and 193, (34) SEQ ID NO: 43, 44, 194, 46, 47 and 193, (35) SEQ ID NO: 175-179 and 195, and (36) SEQ ID NO: 187, 188, 196, 190, 191 and 192.

[0420] In an exemplary embodiment, the FAP antigen-binding protein comprises: VH, which comprises: (i) CDR-H1 comprising any one of SEQ ID NO: 398-402; (ii) CDR-H2 comprising any one of SEQ ID NO: 403-407; and (iii) CDR-H3 comprising any one of SEQ ID NO: 408-412; and VL, which comprises: (i) CDR-L1 comprising any one of SEQ ID NO: 413-417; (ii) CDR-L2 comprising any one of SEQ ID NO: 418-422; and (iii) CDR-L3 comprising any one of SEQ ID NO: 423-427.

[0421] In an exemplary embodiment, the FAP antigen-binding protein comprises: VH, which comprises: (i) CDR-H1 comprising any one of SEQ ID NO: 428-432; (ii) CDR-H2 comprising any one of SEQ ID NO: 433-437; and (iii) CDR-H3 comprising any one of SEQ ID NO: 438-442; and VL, which comprises: (i) CDR-L1 comprising any one of SEQ ID NO: 443-447; (ii) CDR-L2 comprising any one of SEQ ID NO: 448-452; and (iii) CDR-L3 comprising any one of SEQ ID NO: 453-457.

[0422] In an exemplary embodiment, the FAP antigen-binding protein comprises: VH, which comprises: (i) CDR-H1 comprising any one of SEQ ID NO: 458-462; (ii) CDR-H2 comprising any one of SEQ ID NO: 463-467; and (iii) CDR-H3 comprising any one of SEQ ID NO: 468-472; and VL, which comprises: (i) CDR-L1 comprising any one of SEQ ID NO: 473-477; (ii) CDR-L2 comprising any one of SEQ ID NO: 478-482; and (iii) CDR-L3 comprising any one of SEQ ID NO: 483-487.

[0423] In an exemplary embodiment, the FAP antigen-binding protein comprises: VH, which comprises: (i) CDR-H1 comprising any one of SEQ ID NO: 488-492; (ii) CDR-H2 comprising any one of SEQ ID NO: 493-497; and (iii) CDR-H3 comprising any one of SEQ ID NO: 498-502; and VL, which comprises: (i) CDR-L1 comprising any one of SEQ ID NO: 503-507; (ii) CDR-L2 comprising any one of SEQ ID NO: 508-512; and (iii) CDR-L3 comprising any one of SEQ ID NO: 513-517.

[0424] Generally, CDRs are separated by "framework" (FR) residues. The VH or VL framework contains four frame subregions FR1, FR2, FR3, and FR4, with CDRs distributed within these subregions in the following structure: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. Therefore, the antigen-binding proteins described herein can contain both VH frames (such as human VH frame sequences) and VL frames (such as human VL frame sequences).

[0425] Preferred human germline light chain frameworks are those derived from the Vκ or Vλ germline. It should be understood that if a sequence is “derived from” one or more germlines, it refers to a structural relationship where the sequence characteristics correspond to the mentioned germline sequence, but may contain somatic mutations or other amino acid differences relative to the mentioned germline sequence. For sequences “derived from” germlines, the actual process of deriving the sequence from the germline sequence (via molecular biology or computational analysis) is not necessarily required. For example, the VL framework can be derived from one of the following germline frameworks: DPK9 (IMGT name: IGKV1-39), DPK12 (IMGT name: IGKV2D-29), DPK18 (IMGT name: IGKV2-30), DPK24 (IMGT name: IGKV4-1), HK102_V1 (IMGT name: IGKV1-5), DPK1 (IMGT name: IGKV1-33), and DPK8 (IMGT name: IGKV1-9). DPK3 (IMGT name: IGKV1-6), DPK21 (IMGT name: IGKV3-15), Vg_38K (IMGT name: IGKV3-11), DPK22 (IMGT name: IGKV3-20), DPK15 (IMGT name: IGKV2-28), DPL16 (IMGT name: IGLV3-19), DPL8 (IMGT name: IGLV1-40), and V1-22 (IMGT name: IGLV6-57). Alternatively or additionally, frame sequences can be derived from hominin shared frame sequences, such as human Vλ1 shared sequences, Vλ3 shared sequences, Vκ1 shared sequences, Vκ2 shared sequences, and Vκ3 shared sequences. Homininin frame sequences are available from various public databases, such as V-base, IMGT, NCBI, or Abysis.

[0426] The FAP antigen-binding proteins described herein may comprise a VL framework, wherein the framework may contain one or more amino acid substitutions, additions, or deletions, while retaining functional and structural similarity to the lineage from which it is derived. In some aspects, the VL framework is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the human VL framework sequence. In some aspects, antigen-binding proteins, antibodies, or their antigen-binding fragments comprise a VL framework containing 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, additions, or deletions relative to the human VL framework sequence.

[0427] VH frame sequences can be derived from human VH3, VH1, VH5, human VH2, or VH4 lineages. Preferred human lineage heavy chain frames are those derived from VH1, VH3, or VH5 lineages. For example, the VH framework can be derived from one of the following lineages: DP54 or IGHV3-7, DP47 or IGHV3-23, DP71 or IGHV4-59, DP75 or IGHV1-2_02, DP10 or IGHV1-69, DP7 or IGHV1-46, DP49 or IGHV3-30, DP51 or IGHV3-48, DP38 or IGHV3-15, DP79 or IGHV4-39, DP78 or IGHV4-30-4, DP73 or IGHV5-51, DP50 or IGHV3-33, DP46 or IGHV3-30-3, DP31 or IGHV3-9. Alternatively or additionally, the frame sequence may be derived from a common sequence frame, such as: VH3 phylogenetic common sequence, VH1 phylogenetic common sequence, VH5 phylogenetic common sequence, VH2 phylogenetic common sequence or VH4 phylogenetic common sequence.

[0428] The antigen-binding proteins described herein may comprise a VH framework, wherein the framework may contain one or more amino acid substitutions, additions, or deletions, while retaining functional and structural similarity to the lineage from which it is derived. In some aspects, the VH framework is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the human VH framework sequence. In some aspects, the antigen-binding protein, antibody, or antigen-binding fragment thereof comprises a VH framework containing 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acid substitutions, additions, or deletions relative to the human VH framework sequence.

[0429] In an exemplary embodiment, the FAP antigen-binding protein comprises the following: SEQ ID NO: 197, 199, 202, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227, 229, 231, 233, 235, 237, 239, 241, 243, 245, 247, 249, 251, 253, 255, 257, 259, 261, 263, 265, 267, 269, 271, 273, 275, 277, 279, 281, 283, 285, 287, 289, 291, 293, 29 5, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, 325, 327, 329, 331, 332, 334, 335, 339, or 341 at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the same VH. In an exemplary embodiment, the FAP antigen-binding protein comprises the same VH as SEQ ID NO: 198, 200, 202, 204, 206, 208, 210, 212, 214, 216, 218, 220, 222, 224, 226, 228, 230, 232, 234, 236, 238, 240, 242, 244, 246, 248, 250, 252, 254, 256, 258, 260, 262, 264, 266, 268, 270, 272, 274, 276, 278, 280, 282, 284, 286, 288, 290, 292, 294, 29 6, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, 326, 328, 330, 333, 336, 337, 338, 340, or 342, comprising at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical VL sequences. Preferably, the FAP antigen-binding protein comprises a pair of VH and VL sequences listed under the same clone names in Table B.

[0430] In some embodiments, the FAP antigen-binding protein comprises a CH1 domain, preferably a human CH1 domain (such as human IgG1 CH1, human IgG2 CH1, human IgG3 CH1, or human IgG4 CH1). Non-limiting examples of human CH1 sequences are provided in the sequence listing. In some embodiments, the CH1 domain comprises a sequence that is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to SEQ ID NO: 347, 364, 368, or 372.

[0431] In some embodiments, the FAP antigen-binding protein described herein includes an Fc domain. The Fc domain may be derived from IgA (e.g., IgA1 or IgA2), IgG, IgE, or IgG (e.g., IgG1, IgG2, IgG3, or IgG4). In some embodiments, the Fc domain includes a wild-type sequence of the Fc domain. Non-limiting examples of human Fc sequences are provided in the sequence listing.

[0432] In some embodiments, the Fc domain contains one or more mutations that result in alterations in biological activity, such as improving half-life / stability or making the antibody more suitable for expression / manufacturability. For example, mutations can be introduced into the Fc domain to reduce effector activity (e.g., WO 2005 / 063815) and / or increase homogeneity during recombinant protein production.

[0433] Generally, the amino acid residues in the constant domain of the IgG heavy chain of an antibody are numbered according to the EU index (as described in Kabat et al., 1991), as described in Edelman et al., 1969, Proc. Natl. Acad. Sci. USA [Proceedings of the National Academy of Sciences] 63(1):78-85, referred to herein as the “EU index number”. Typically, the constant domain contains residues 118 to 447 of the human IgG1 constant domain, and the Fc domain contains residues 236 to 447. Comparisons between EU numbers and other numbering systems can be found, for example, in the IGMT database.

[0434] The amino acid residues of the constant domain of the light chain are numbered according to Kabat et al., 1991, "Sequences of Proteins of Immunological Interest, 5th Edition", NATIONALINSTITUTES OF HEALTH. The κ light chain also has an EU index number, and the EU index is the same as that of Kabat. The λ light chain does not have an EU index number.

[0435] In some embodiments, the Fc domain is the Fc domain of human IgG1 and contains one or more of the following effector-null mutations: L234A, L235A, and G237A (according to EU index number), commonly referred to as the “LALA” mutation.

[0436] A single mutation in L235E has been reported to be sufficient to knock out the binding of the Fc receptor on U937 cells. Furthermore, the 100-fold reduction in binding to FcγR in the presence of L235E-mutant IgG1 also resulted in lower T cell activation and proliferation. Based on this initial mutation, a combination of L234A and L235A (commonly referred to as the LALA mutation) has been found to eliminate FcγRIIa binding. These two mutations have later been shown to eliminate detectable binding of both IgG1 and IgG4 to FcγRI, IIa, and IIIa. Other site mutations (such as Gly237Ala, Glu318Ala, Asp265Ala, and Glu233Pro mutations) have been reported to knock out Fc receptor binding.

[0437] In an exemplary embodiment, the Fc region contains a stable effector-free function (SEFL) mutation to reduce the ability to interact with the Fcγ receptor. SEFL mutations are known in the art. See, for example, Liu et al., J Biol Chem 292: 1876-1883 (2016); and Jacobsen et al., J Biol. Chem. 292:1865-1875 (2017). Furthermore, US US9546203 discloses an Fc region containing an N297G mutation and one or more substitutions of cysteine ​​amino acid residues at positions V259, A287, R292, V302, L306, V323, or I332 (using the EU numbering scheme). In an exemplary aspect, the SEFL mutation includes one or more of the following mutations (according to EU system numbers): L242C, A287C, R292C, N297G, V302C, L306C, and / or K334C. In an exemplary aspect, the SEFL mutation includes N297G. In an exemplary aspect, the SEFL mutation includes A287C, N297G, and L306C. In other exemplary aspects, the SEFL mutation includes R292C, N297G, and V302C (i.e., SEFL2-2).

[0438] In an exemplary embodiment, the Fc region contains the YTE mutation. The M252Y / S254T / T256E (EU index number, referred to as "YTE") triple mutation has been shown to increase the IgG half-life by approximately 4-fold in cynomolgus monkeys.

[0439] C-terminal lysine cleavage is a common phenomenon occurring during the biological production of monoclonal antibodies. Typically, lysine residues are removed via carboxypeptidase D (CpD), resulting in a mixture of antibody isotypes with zero or one C-terminal lysine residue on each heavy chain. Further, following C-terminal lysine cleavage, peptidyl-glycine α-amidyl-monooxygenase (PAM) catalyzes the hydroxylation of glycine and the removal of glyoxylate from the glycine residue, leaving an amidated C-terminal proline. Therefore, during recombinant monoclonal antibody production, the product is typically a mixture of C-terminal processed variants at the (amidated) proline, glycine, or lysine of the heavy chain. Sometimes, the deletion of the C-terminal lysine with the Fc domain may be desirable to increase homogeneity during recombinant protein production.

[0440] In some embodiments, terminal lysine may be absent; in some embodiments, terminal lysine may be present; in some embodiments, terminal glycine-lysine may be absent; in some embodiments, terminal glycine-lysine may be present.

[0441] In exemplary embodiments, the FAP antigen-binding protein described herein comprises an Fc derived from IgG1. In some embodiments, the Fc comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO: 348, 355, 356, 357, 358, 359, 360, 366, 370, or 374.

[0442] In exemplary embodiments, the FAP antigen-binding protein described herein comprises an IgG1 heavy chain constant domain. In some embodiments, the heavy chain constant domain comprises at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 90%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 97%, at least 98%, at least 99%, or 100% identical to the sequence in SEQ ID NO: 349, 350, 351, 352, 353, 354, 361, 362, 363, 367, 371, or 375.

[0443] In some embodiments, the FAP antigen-binding protein described herein comprises a κ or λ light chain constant domain. In some embodiments, the κ light chain constant domain comprises a sequence that is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to that in SEQ ID NO:343 or 344. In some embodiments, the λ light chain constant domain comprises a sequence that is at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 86%, at least 87%, at least 88%, at least 89%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% identical to that in SEQ ID NO:345 or 346.

[0444] FAP antigen-binding proteins can be full-length immunoglobulins, Fab, or scFv. scFv may contain a linker between VH and VL. Exemplary linker sequences, such as GS-based linkers, are provided in the sequence listing.

[0445] 2.4 Bispecific molecules targeting FAP.

[0446] In various respects, an antigen-binding protein is a bispecific molecule that binds to two different antigens or targets. In various cases, the bispecific molecule binds to both FAP and T-cell co-stimulatory molecules. In an exemplary aspect, the bispecific molecule comprises four antigen-binding sites, two of which bind to the FAP protein and two of which bind to the T-cell co-stimulatory molecule. Optionally, each FAP binding site is identical to the other FAP binding site.

[0447] In one specific instance, a bispecific molecule comprises both an IgG moiety and an scFv moiety. For example... Figure 5 The results show the presence of two Fab moieties that bind to an antigen (e.g., a T-cell co-stimulatory molecule). Each Fab moieties comprises two chains: one containing a heavy chain variable domain A (VH). A Heavy chains containing the CH1 and CH1 domains, and heavy chains containing the variable domain A (VL) of the light chain. A The Fab and CL domains are light chains. Each Fab is linked to one chain of Fc (CH2-CH3) to form IgG. Because this part of the structure is essentially IgG, there are no new linkers between Fab and Fc (Fab and Fc are linked by a "hinge" sequence, just like wild-type IgG). Furthermore, there are two scFv portions that bind to another antigen (e.g., FAP). Each scFv contains a heavy chain variable domain B (VH2-CH3). B ) and light chain variable structural domain B (VL B ); and VH B and VL B The first linker connects the IgG molecules. Then, the second linker connects the C-terminus of a CH3 domain to the N-terminus of an scFv. This particular structure is sometimes referred to as the "IgG-scFv" form (one or more scFv portions are attached to an IgG molecule). Because each target has two binding domains, Figure 5 The bispecific molecules illustrated herein are generally referred to as “bivalent” bispecific molecules; however, it should be noted that it is also acceptable in the art to refer to such molecules as “tetravalent”, since there are a total of four binding domains.

[0448] It is believed that compared to monovalent binding of FAP (e.g., a heterologous Ig molecule in which one arm of the antibody binds to a T-cell costimulatory molecule and the other arm of the antibody binds to FAP), bivalent binding of FAP can enhance the cross-linking (clustering) of T-cell costimulatory molecules and potentially enhance the overall potency of the molecule.

[0449] A "linker" is a molecule or group of molecules that connects two separate entities to each other and provides space and flexibility between the two entities, enabling them to achieve conformations in which they, for example, specifically bind their corresponding substances. Protein linkers are particularly preferred and can be expressed as components of recombinant proteins using standard recombinant DNA techniques well known in the art. For recombinant proteins comprising two or more linkers as described herein (e.g., the IgG-scFv form), the linkers may all be identical, or some or all of the linkers may be different from each other.

[0450] In some embodiments, the linker is a peptide linker. In some embodiments, the peptide linker comprises about 1 to 30 amino acid residues. Exemplary linkers include, for example, a glycine-rich peptide; a peptide comprising glycine and serine; a peptide having the sequence [Gly-Gly-Ser]n (SEQ ID NO: 388), wherein n is 1, 2, 3, 4, 5, or 6; or a peptide having the sequence [Gly-Gly-Gly-Gly-Ser]n (SEQ ID NO: 387), wherein n is 1, 2, 3, 4, 5, or 6. A glycine-rich peptide linker comprises a peptide linker in which at least 25% of the residues are glycine. Glycine-rich peptide linkers are well known in the art (e.g., Chichili et al. Protein Sci. [Protein Science] Feb. 2013; 22(2): 153-167). The peptide linker may also be a proline-threonine-rich peptide linker.

[0451] like Figure 5 The results show that when a bispecific molecule contains the scFv moiety, mutations can be introduced into the scFv to further improve its stability. For example, it has been reported that insufficient interfacial stability between the heavy and light chains of the scFv fragment may be a major cause of irreversible scFv inactivation. It has been reported that the Fv fragment has a 10... -9 Up to 10 -6 K in the range of M DThe cysteine ​​residue dissociates into a heavy-chain variable domain (VH) and a light-chain variable domain (VL). Disulfide bonds between these domains have been used to further enhance the stability of the scFv. For example, mutations to Cys at H44 (Kabat number) and L100 (Kabat number) do not significantly affect domain folding. These two cysteine ​​residues can then form an intramolecular disulfide bond to further stabilize the scFv. Such mutations are sometimes referred to as "cysteine ​​clamps."

[0452] Sequence listing B shows a specific instance of scFv containing a cysteine ​​clamp, where mutations at H44 (Kabat number) and L100 (Kabat number) are used to generate disulfide bonds (called "CC").

[0453] In an exemplary embodiment, the scFv portion is bound to FAP and contains at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% of the same amino acid sequence as SEQ ID NO:393, 518, 519, 520, or 521.

[0454] The bispecific molecules disclosed in this article may further include CL, CH1 and / or Fc regions (as described in detail above).

[0455] 2.5 Combination Features The antigen-binding proteins provided herein bind to their respective targets or antigens in a non-covalent and reversible manner. In exemplary embodiments, the binding strength of the antigen-binding protein to its target or antigen (e.g., FAP) can be described by its affinity, which is a measure of the strength of the interaction between the binding site of the antigen-binding protein and the target or antigen (e.g., FAP). In an exemplary aspect, the antigen-binding proteins provided herein have a high affinity for their targets or antigens (e.g., FAP), and therefore will bind a larger amount of the target or antigen (e.g., FAP) in a shorter time period than low-affinity antigen-binding proteins. In an exemplary aspect, the antigen-binding proteins provided herein have a low affinity for FAP, and therefore will bind a smaller amount of FAP in a longer time period than high-affinity antigen-binding proteins. In an exemplary aspect, the equilibrium association constant KA of the antigen-binding protein is at least 10. 5 M -1 At least 10 6 M -1 At least 10 7 M -1 At least 10 8 M -1 At least 109 M -1 At least 10 10 M -1 At least 10 11 M -1 At least 10 12 M -1 At least 10 13 M -1 Or at least 10 14 M -1 As is known to those skilled in the art, KA can be affected by factors including pH, temperature, and buffer composition.

[0456] In an exemplary embodiment, the binding strength of an antigen-binding protein to its target or antigen (e.g., FAP) can be described by its sensitivity. D The equilibrium dissociation constant (k) between an antigen-binding protein and its target or antigen (e.g., FAP). off / k on (ratio). K D It is inversely correlated with KA. K D The value is related to the concentration of antigen-binding protein (the amount of antigen-binding protein required for a specific experiment), therefore K D The lower the value (the lower the required concentration), the higher the affinity of the antigen-binding protein. In an exemplary aspect, the binding strength of an antigen-binding protein to its target (e.g., FAP) can be expressed using KA. D To describe. In an exemplary aspect, the K antigen-binding protein provided herein... D The value is approximately 10 -1 M or smaller, approximately 10 -2 M or smaller, approximately 10 -3 M or smaller, approximately 10 -4 M or smaller, approximately 10 -5 M or smaller, approximately 10 -6 M or smaller, approximately 10 -7 M or smaller, approximately 10 -8 M or smaller, approximately 10 -9 M or smaller, approximately 10 -10 M or smaller, approximately 10 -11 M or smaller, approximately 10 -12 M or smaller, approximately 10 -13 M or smaller, approximately 10 -14 M or smaller, approximately 10 -5 M to approximately 10 -15 M, approximately 10 -6 M to approximately 10 -15 M, approximately 10 -7 M to approximately 10 -15 M, approximately 10-8 M to approximately 10 -15 M, approximately 10 -9 M to approximately 10 -15 M, approximately 10 -10 M to approximately 10 -15 M, approximately 10 -5 M to approximately 10 -14 M, approximately 10 -6 M to approximately 10 -14 M, approximately 10 -7 M to approximately 10 -14 M, approximately 10 -8 M to approximately 10 -14 M, approximately 10 -9 M to approximately 10 -14 M, approximately 10 -10 M to approximately 10 -14 M, approximately 10 -5 M to approximately 10 -13 M, approximately 10 -6 M to approximately 10 -13 M, approximately 10 -7 M to approximately 10 -13 M, approximately 10 -8 M to approximately 10 -13 M, approximately 10 -9 M to approximately 10 -13 M, or approximately 10 -10 M to approximately 10 -13 M.

[0457] In an exemplary aspect, the K antigen-binding protein provided herein D The values ​​are in micromolar, nanomolar, picomolar, or femtomolar quantities. In an exemplary aspect, the K-type antigen-binding protein provided herein... D In about 10 -4 Up to 10 -6 M, or 10 -7 Up to 10 -9 M, or 10 -10 Up to 10 -12 M, or 10 -13 Up to 10 -15 Within the M range. In an exemplary aspect, the antigen-binding protein has a Kk of approximately 0.07 nM to approximately 4 nM. DThe antigen-binding protein binds to human FAP at values ​​of approximately 0.01 nM to approximately 50 nM, approximately 0.02 nM to approximately 50 nM, approximately 0.05 nM to approximately 50 nM, approximately 0.05 nM to approximately 45 nM, approximately 0.05 nM to approximately 40 nM, approximately 0.05 nM to approximately 35 nM, approximately 0.05 nM to approximately 30 nM, approximately 0.05 nM to approximately 25 nM, approximately 0.05 nM to approximately 20 nM, approximately 0.05 nM to approximately 15 nM, or approximately 0.05 nM to approximately 10 nM. D Binds to human FAP. In an exemplary aspect, the antigen-binding protein is expressed at a K+ level of approximately 0.05 nM to approximately 4 nM. D It binds to FAP in cynomolgus monkeys. In an exemplary aspect, the antigen-binding protein is expressed in K at concentrations of approximately 0.01 nM to approximately 50 nM, approximately 0.02 nM to approximately 50 nM, approximately 0.05 nM to approximately 50 nM, approximately 0.05 nM to approximately 45 nM, approximately 0.05 nM to approximately 40 nM, approximately 0.05 nM to approximately 35 nM, approximately 0.05 nM to approximately 30 nM, approximately 0.05 nM to approximately 25 nM, approximately 0.05 nM to approximately 20 nM, approximately 0.05 nM to approximately 15 nM, or approximately 0.05 nM to approximately 10 nM. D Binding to FAP in cynomolgus monkeys. In an exemplary embodiment, the FAP antigen-binding protein is expressed at a K+ level of approximately 0.05 nM to approximately 5 nM. D The value binds to human FAP. In an exemplary embodiment, the FAP antigen-binding protein is expressed at a Kc concentration of approximately 0.05 nM to approximately 5 nM. D The value is associated with the FAP of cynomolgus monkeys.

[0458] K can be determined using methods established in the art. D Value. Used to measure K. D One exemplary method is surface plasmon resonance (SPR), a method well-known in the art (e.g., Nguyen et al., Sensors [Sensors] (Basel). 2015 May 5; 15(5):10481-510). K can be measured via SPR using biosensor systems such as the BIACORE® system. D Value. BIAcore kinetic analysis includes analyzing the binding and dissociation of antigens with chips having immobilized molecules (e.g., molecules containing epitope-binding domains) on their surfaces. This art is used to determine the K-value of proteins. DAnother well-known method is to use biolayer interferometry (e.g., Shah et al. J Vis Exp. [Video Experiment Journal] 2014;(84): 51383). K can be measured by biolayer interferometry using OCTET® technology (Octet QKe system, ForteBio). D The value can be determined alternatively or additionally using a KinExA® (kinetic repulsion assay) available from Savidyne Instruments (Boise, Idaho). This document covers any methods known in the art for assessing binding affinity between two binding partners.

[0459] In some respects, K D The values ​​are measured via surface plasmon resonance (SPR). Antigens (e.g., FAP) can be immobilized on, for example, a solid surface. Antigens can also be immobilized on a chip, for example, via covalent coupling (e.g., amine coupling). The chip can be a CM5 sensor chip. As the analyte binds to the ligand, the accumulation of protein on the sensor surface leads to an increase in refractive index. This refractive index change is measured in real time (sampled every 0.1 s in kinetic analysis experiments), and the results are plotted relative to time in units of response (RU) (called a sensor plot). If there is a difference in refractive index between the run buffer and the sample buffer, a response (background response) will also be generated. This background response must be subtracted from the sensor plot to obtain the actual binding response. The background response is recorded by injecting the analyte through a control or reference flow cell that does not have ligands immobilized on the sensor surface or unrelated ligands. Real-time measurements of the association and dissociation of binding interactions allow for the calculation of association and dissociation rate constants, as well as the corresponding affinity constants. One RU represents 1 pg of protein bound per square mm. In practice, an analyte binding density of more than 50 pg per square mm is typically required to generate a good reproducible response.

[0460] It can monitor the dissociation of antigen-binding proteins from antigens for approximately 3600 seconds. SPR analysis and data collection can be performed and conducted at temperatures ranging from approximately 15°C to approximately 37°C. SPR analysis and data collection can be performed and conducted at approximately 25°C to 37°C. SPR analysis and data collection can be performed and conducted at approximately 37°C. SPR analysis and data collection can be performed and conducted at 37°C. K D The values ​​can be measured using the BIAcore T200 instrument via SPR. SPR rate and affinity can be determined by fitting the obtained sensor data to a 1:1 model in BIAcore T200 evaluation software version 1.0. The acquisition rate can be approximately 1 Hz.

[0461] Used to determine antibody K DAnother method is to use biolayer interferometry (BLI), typically performed using OCTET® technology (Octet QKe system, Pallford Biosciences). In some embodiments, biosensor analysis is used. Typically, one interacting element (“ligand,” such as an antigen-binding protein) is immobilized on the surface of the biosensor, and another interacting element (“analyte,” such as an antigen) is retained in solution. The assay begins with an initial baseline or equilibration step using an assay buffer. Next, the ligand (such as an antigen-binding protein) is immobilized on the surface of the biosensor by direct immobilization or a capture-based method (loading). After ligand immobilization, the biosensor is immersed in a buffer solution for a baseline step to assess assay drift and determine the ligand loading level. After the baseline step, the biosensor is immersed in a solution containing the binding partner of the ligand (analyte) (association). In this step, the binding interaction between the analyte and the immobilized ligand is measured. After analyte association, the biosensor is immersed in a buffer solution without analyte, allowing the bound analyte to detach from the ligand (dissociation). The assay sequence is then repeated on a new or regenerated biosensor for each tested analyte. Each binding response is measured and reported in real time on the sensor trace. The instrument can be an Octet QKe system, Octet RED96 system, Octet QK384 system, or RED384 system.

[0462] In some embodiments, FAP-binding proteins are at a K value equal to or less than the following: D Values ​​combined with human FAP: approximately 200 nM, approximately 150 nM, approximately 100 nM, approximately 90 nM, approximately 80 nM, approximately 70 nM, approximately 60 nM, approximately 50 nM, approximately 40 nM, approximately 30 nM, approximately 25 nM, approximately 20 nM, approximately 15 nM, approximately 10 nM, approximately 9 nM, approximately 8 nM, approximately 7 nM, approximately 6 nM, approximately 5 nM, approximately 4 nM, approximately 3 nM, approximately 2 nM, approximately 1 nM, approximately 900 pM, approximately 800 pM, approximately 700 pM, approximately 600 pM, approximately 500 pM, approximately 400 pM, approximately 300 pM, approximately 250 pM, approximately 200 pM, approximately 150 pM, approximately 100 pM, approximately 50 pM, approximately 40 pM, approximately 30 pM, approximately 25 pM, approximately 20 pM, approximately 15 pM, approximately 10 pM, approximately 5 pM, or approximately 1 pM. K D The value can be measured by surface plasmon resonance (SPR) (e.g., Biacore T200 instrument); or it can be measured by biolayer interferometry (BLI) (e.g., ForteBio Octet instrument).

[0463] Examples 1 and 2 also provide exemplary methods for measuring the binding of antigen-binding proteins to their targets.

[0464] 2.6 Cross-reactivity In each respect, the antigen-binding protein binds to human FAP. A reference amino acid sequence of human FAP is provided herein as SEQ ID NO: 394 (the recombinant expressed soluble FAP is provided as SEQ ID NO: 395). In each respect, the antigen-binding protein binds to cynoma (cyno) FAP. The amino acid sequence of cyno FAP is provided herein as SEQ ID NO: 396. In an exemplary respect, the antigen-binding protein binds with high affinity to both human FAP and cyno FAP. In various embodiments, the antigen-binding protein disclosed herein binds to both human FAP and cyno FAP, but does not cross-react with any other FAP orthologs. In various cases, the antigen-binding protein binds with high affinity to both human FAP and cyno FAP, but does not bind to any other FAP orthologs, e.g., not to mouse FAP, rat FAP, canine FAP, bovine FAP, etc. In various embodiments, the selectivity of the disclosed antigen-binding protein for human FAP and cyno FAP is at least 10-fold, 5-fold, 4-fold, 3-fold, and 2-fold greater than the selectivity of the antigen-binding protein for any other FAP ortholog. In various embodiments, the selectivity of the disclosed antigen-binding protein for any other FAP ortholog is K... D This antigen-binding protein has a K+ level against human FAP and cyno FAP. D At least 10 times, 5 times, 4 times, 3 times, or 2 times.

[0465] 2.7 Competition Measurement In various embodiments, the antigen-binding protein inhibits the binding interaction between human FAP and a reference antibody known to bind to FAP. For example, the reference antibody may be a FAP antigen-binding protein disclosed herein, such as the FAP antigen-binding protein disclosed in the sequence listing. In various cases, the FAP antigen-binding protein competitively binds to the reference antibody, thereby reducing the amount of human FAP bound to the reference antibody, as determined by an in vitro competitive binding assay. In all aspects, the FAP antigen-binding protein inhibits the binding interaction between human FAP and the reference antibody, and this inhibition is measured using IC50. 50 Characterization. In all aspects, the FAP antigen-binding protein exhibited an IC50 of less than approximately 2500 nM for inhibiting the binding interaction between human FAP and the reference antibody. 50In various aspects, antigen-binding proteins exhibit IC50 values ​​of less than approximately 2000 nM, less than approximately 1500 nM, less than approximately 1000 nM, less than approximately 900 nm, less than approximately 800 nm, less than approximately 700 nm, less than approximately 600 nm, less than approximately 500 nm, less than approximately 400 nm, less than approximately 300 nm, less than approximately 200 nm, or less than approximately 100 nm. 50 In all aspects, the antigen-binding protein exhibited IC50 values ​​of less than approximately 90 nM, less than approximately 80 nM, less than approximately 70 nM, less than approximately 60 nM, less than approximately 50 nM, less than approximately 40 nM, less than approximately 30 nM, less than approximately 20 nM, or less than approximately 10 nM. 50 In various cases, the FAP antigen-binding protein competes with the reference antibody for binding to human FAP, thereby reducing the amount of human FAP bound to the reference antibody, as determined by FACS-based assays (where the fluorescence of a fluorophore-conjugated secondary antibody bound to the Fc of the reference antibody is measured in the absence or presence of a specific amount of FAP antigen-binding protein). In each case, the FACS-based assay is performed using a reference antibody, a fluorophore-conjugated secondary antibody, and cells expressing FAP. In each case, the cells are genetically engineered to overexpress FAP. In some cases, the cells are HEK293T cells transduced with a viral vector to express FAP. Alternatively, the cells endogenously express FAP. In some cases, cells endogenously expressing FAP are pre-defined as low- or high-FAP-expressing cells prior to the FACS-based assay.

[0466] Other binding assays (e.g., competitive binding assays or competitive assays) that test the ability of an antigen-binding molecule (such as an antibody) to compete with a second antigen-binding molecule (such as a second antibody) for binding to an antigen or its epitope are known in the art. See, for example, Trikha et al., Int J Cancer [International Journal of Cancer] 110: 326-335 (2004); Tam et al., Circulation [Circulation] 98(11): 1085-1091 (1998); US Patent Application Publication No. US 20140178905; Chand et al., Biologicals [Biological Products] 46: 168-171 (2017); Liu et al., Anal Biochem [Analytical Biochemistry] 525: 89-91 (2017); and Goolia et al., J Vet Diagn Invest [Veterinary Diagnostic Research Journal] 29(2): 250-253 (2017). In addition, other methods for comparing two antigen-binding molecules are known in the art, including, for example, surface plasmon resonance (SPR). SPR can be used to determine the binding constants of two binding molecules and to compare these two binding constants.

[0467] 3. Nucleic acids, vectors, and host cells 3.1 Nucleic Acid This disclosure further provides nucleic acids comprising the nucleotide sequence encoding the antigen-binding protein disclosed herein. Nucleic acids may comprise a single nucleic acid molecule, or two or more nucleic acid molecules (e.g., a first nucleic acid molecule encoding a heavy chain amino acid sequence and a second nucleic acid molecule encoding a light chain amino acid sequence). In some respects, the nucleic acids disclosed herein are recombinant.

[0468] Furthermore, the nucleic acid sequence encoding the signal peptide can be added to the 5' of the polypeptide disclosed herein. Recombinant expression of the antigen-binding protein (or antigen-binding moiety) disclosed herein typically requires the molecular energy to be secreted. The translocation of nascent proteins from the cytoplasm to the ER mediated by their signal peptide is a crucial step in protein secretion. It should be understood that the signal peptide is present (and is often critical) during the initial synthesis of the nascent protein, but is subsequently cleaved during the secretion process. Therefore, although mature proteins no longer possess the signal peptide, the presence of a signal peptide-encoding sequence in the nucleic acid is generally essential for recombinant protein expression.

[0469] 3.2 Carrier In some aspects, nucleic acids disclosed herein are introduced into vectors. In this regard, this disclosure provides vectors comprising any nucleic acid disclosed herein. In an exemplary aspect, the vector is a recombinant expression vector. For purposes herein, the term "recombinant expression vector" means a genetically modified oligonucleotide or polynucleotide construct that, when the construct contains a nucleotide sequence encoding an mRNA, protein, polypeptide, or peptide, allows the cell to express that mRNA, protein, polypeptide, or peptide when the vector is contacted with a host cell under conditions sufficient to allow intracellular expression of that mRNA, protein, polypeptide, or peptide. The vectors disclosed herein are generally non-natural. However, a portion of the vector may be naturally occurring. The vectors disclosed herein may contain any type of nucleotide, including but not limited to DNA and RNA, which may be single-stranded or double-stranded, synthetic or partially derived from natural sources, and may contain natural, non-natural, or modified nucleotides. The vector may contain naturally occurring internucleotide bonds, or non-natural internucleotide bonds, or both. In some aspects, modified nucleotides or non-natural internucleotide bonds do not impede transcription or replication of the vector.

[0470] The vectors disclosed herein can be any suitable vector and can be used to transform or transfect any suitable host. Suitable vectors include those designed for propagation and amplification, or for expression, or both, such as plasmids and viruses. Vectors can be selected from the following groups: pUC series (Fermentas LifeSciences), pBluescript series (Stratagene, La Jolla, California), pET series (Novagen, Madison, Wisconsin), pGEX series (Pharmacia Biotech, Uppsala, Sweden), and pEX series (Clontech, Palo Alto, California). Phage vectors such as λGTIO, λGTl 1, λZapII (Stratagene), λEMBL4, and λNMl149 can also be used. Examples of plant expression vectors include pBI101, pBI101.2, pBI101.3, pBI121, and pBIN19 (Crotec). Examples of animal expression vectors include pEUK-Cl, pMAM, and pMAMneo (Crotec). In some respects, the vectors are viral vectors, such as retroviral vectors.

[0471] The vectors disclosed herein can be prepared using standard recombinant DNA techniques, such as those described in Sambrook et al., ibid. and Ausubel et al., ibid. Constructs of circular or linear expression vectors containing a replication system that functions in prokaryotic or eukaryotic host cells can be prepared. The replication system can be derived from, for example, CoIEl, 2 μ plasmid, λ, SV40, bovine papillomavirus, etc.

[0472] In some respects, vectors contain regulatory sequences (such as transcription and translation start and stop codons) that are specific to the type of host to which the vector is to be introduced (e.g., bacteria, fungi, plants, or animals) (depending on whether the vector is DNA-based or RNA-based).

[0473] The vector may include one or more biomarker genes that allow selection of the host for transformation or transfection. Biomarker genes include biocidal resistance (e.g., resistance to antibiotics, heavy metals, etc.), complementation in auxotrophic hosts to provide protrophic types, etc. Biomarker genes suitable for the expression vectors disclosed herein include, for example, neomycin / G418 resistance genes, hygromycin resistance genes, histidine resistance genes, tetracycline resistance genes, and ampicillin resistance genes.

[0474] The vector may contain a natural or standard promoter of a nucleotide sequence operatively linked to a nucleotide sequence encoding a polypeptide (including its functional moiety and functional variants) or a nucleotide sequence complementary to or hybridized to a nucleotide sequence encoding an antigen-binding protein. The selection of promoters, for example, strong, weak, inducible, tissue-specific, and developmentally specific, is within the general skill of a person skilled in the art. Similarly, the combination of nucleotide sequences and promoters is also within the skill of a person skilled in the art. Promoters may be non-viral or viral promoters, such as cytomegalovirus (CMV) promoters, SV40 promoters, RSV promoters, and promoters found in long terminal repeat sequences of mouse stem cell viruses.

[0475] 3.3 Host Cell This document provides host cells containing the nucleic acids or vectors disclosed herein. As used herein, the term "host cell" refers to any type of cell that may contain the vectors disclosed herein and is capable of producing expression products encoded by nucleic acids (e.g., mRNA, proteins). In some aspects, host cells are adherent cells or suspension cells, i.e., cells that grow in suspension. In exemplary aspects, host cells are cultured cells or primary cells, i.e., cells directly isolated from an organism (e.g., humans). Host cells may belong to any cell type, may originate from any type of tissue, and may be at any developmental stage.

[0476] In an exemplary aspect, the cell is a eukaryotic cell, including but not limited to yeast cells, filamentous fungal cells, protozoan cells, algal cells, insect cells, or mammalian cells. Such host cells are described in the art. See, for example, Frenzel et al., Front Immunol 4: 217 (2013). In an exemplary aspect, the eukaryotic cell is a mammalian cell. In an exemplary aspect, the mammalian cell is a non-human mammalian cell. In some respects, the cells are Chinese ovarian (CHO) cells and their derivatives (e.g., CHO-K1, CHO pro-3, CS9), mouse myeloma cells (e.g., NS0, GS-NS0, Sp2 / 0), cells engineered to lack dihydrofolate reductase (DHFR) activity (e.g., DUKX-X11, DG44), human embryonic kidney 293 (HEK293) cells or their derivatives (e.g., HEK293T, HEK293-EBNA), African green monkey kidney cells (e.g., COS cells, VERO cells), human cervical cancer cells (e.g., HeLa), human osteosarcoma epithelial cells U2-OS, adenocarcinoma human alveolar basal epithelial cells A549, human fibrosarcoma cells HT1080, mouse brain tumor cells CAD, embryonic cancer cells P19, and mouse embryonic fibroblasts NIH. 3T3, mouse fibroblasts L929, mouse neuroblastoma cells N2a, human breast cancer cells MCF-7, retinoblastoma cells Y79, human retinoblastoma cells SO-Rb50, human liver cancer cells HepG2, mouse B myeloma cells J558L, or young hamster kidney (BHK) cells (Gaillet et al. 2007; Khan, Adv PharmBull [Advanced Drug Bulletin] 3(2): 257-263 (2013)). In a particular embodiment, the host cell was CS9 (CHO cell line).

[0477] For the purpose of amplifying or replicating vectors, in some respects the host cell is a prokaryotic cell, such as a bacterial cell.

[0478] This disclosure also provides a cell population comprising at least one host cell as described herein. In some aspects, the cell population is a heterogeneous population comprising host cells containing the vector, and additionally comprising at least one other cell that does not contain any vector. Alternatively, in some aspects, the cell population is a substantially homogeneous population, wherein the population primarily comprises (e.g., is substantially composed of) host cells containing the vector. In some aspects, the population is a clonal population of cells, wherein all cells in the population are clones of a single host cell containing the vector, such that all cells in the population contain the vector. In exemplary embodiments of this disclosure, the cell population is a clonal population comprising host cells containing the vector as described herein.

[0479] 3.4. Manufacturing Method The antigen-binding proteins disclosed herein can be obtained by methods known in the art. Suitable methods for de novo synthesis of peptides are described, for example, in: Chan et al., Fmoc Solid Phase Peptide Synthesis, Oxford University Press, Oxford, 2005; Peptide and Protein Drug Analysis, edited by Reid, R., Marcel Dekker, Inc., 2000; Epitope Mapping, edited by Westwood et al., Oxford University Press, Oxford, 2000; and U.S. Patent No. 5,449,752. Further exemplary methods for preparing the peptides of the present invention are described herein.

[0480] Furthermore, in some respects, the antigen-binding proteins disclosed herein are generated using standard recombination methods on nucleic acids encoding the amino acid sequences of the molecules. See, for example, Sambrook et al., Molecular Cloning: A Laboratory Manual, 3rd ed., Cold Spring Harbor Press, Cold Spring Harbor, NY 2001; and Ausubel et al., Current Protocols in Molecular Biology, Greene Publishing Associates and John Wiley & Sons, NY 1994.

[0481] This document further provides a method for preparing the antigen-binding protein disclosed herein. In an exemplary embodiment, the method includes culturing the host cells disclosed herein to express the antigen-binding protein, and harvesting the expressed antigen-binding protein. The host cell may be any host cell described herein. In an exemplary aspect, the host cell is selected from the group consisting of CHO cells, NSO cells, COS cells, VERO cells, and BHK cells. In an exemplary aspect, the step of culturing the host cell includes culturing the host cell in a growth medium to support the growth and expansion of the host cell. In an exemplary aspect, the growth medium increases cell density, culture viability, and productivity in a timely manner. In an exemplary aspect, the growth medium contains amino acids, vitamins, inorganic salts, glucose, and serum as sources of growth factors, hormones, and adhesion factors. In an exemplary aspect, the growth medium is a culture medium with a fully defined chemical composition consisting of amino acids, vitamins, trace elements, inorganic salts, lipids, and insulin or insulin-like growth factor. In addition to nutrients, the growth medium also helps maintain pH and osmotic pressure. Several growth media are commercially available and described in the art. See, for example, Arora, “Cell Culture Media: A Review” MATER METHODS 3:175 (2013).

[0482] In an exemplary aspect, the method for preparing the antigen-binding protein disclosed herein includes culturing host cells in a fed-batch medium. In an exemplary aspect, the method includes culturing in a fed-batch medium. Methods for producing recombinant proteins are known in the art. See, for example, Li et al., “Cell culture processes for monoclonal antibody production” MAbs [Monoclonal Antibodies] 2(5): 466–477 (2010).

[0483] Methods for preparing the antigen-binding proteins disclosed herein may include one or more steps for purifying the molecule from a cell culture or its supernatant, and preferably recovering the purified protein. In an exemplary aspect, the method includes one or more chromatographic steps, such as affinity chromatography (e.g., protein A affinity chromatography), ion exchange chromatography, or hydrophobic interaction chromatography. In an exemplary aspect, the method includes purifying the protein using a protein A affinity chromatography resin.

[0484] In an exemplary embodiment, the method further includes the step of formulating purified proteins to obtain a formulation containing purified proteins. Such steps are described, for example, in *Formulation and Process Development Strategies for Manufacturing*, edited by Jameel and Hershenson, John Wiley & Sons, Inc. (Hoboken, NJ), 2010.

[0485] 4. Pharmaceutical compositions and treatment methods 4.1 Pharmaceutical Composition This document provides compositions comprising antigen-binding proteins, nucleic acids, vectors, host cells, or combinations thereof. The compositions may comprise isolated and / or purified forms of antigen-binding proteins, nucleic acids, vectors, or host cells, or combinations thereof.

[0486] In an exemplary aspect, the composition comprises an agent that enhances the chemophysical characteristics of an antigen-binding molecule, nucleic acid, carrier, or host cell or a combination thereof in a manner such as by stabilizing the antigen-binding protein, for example, at a specific temperature (e.g., room temperature), increasing the shelf life of the antigen-binding protein, reducing the degradation of the antigen-binding protein (e.g., degradation mediated by oxidative proteases), increasing the half-life of the antigen-binding protein, etc.

[0487] In an exemplary aspect of this disclosure, the composition further comprises a pharmaceutically acceptable carrier, diluent, or excipient. Pharmaceutical compositions may contain any pharmaceutically acceptable ingredients, including, for example, acidifiers, additives, adsorbents, aerosol propellants, air displacement agents, alkalizers, anti-caking agents, anticoagulants, antimicrobial preservatives, antioxidants, antibacterial agents, alkalis, binders, buffers, chelating agents, coating agents, colorants, desiccants, detergents, diluents, disinfectants, disintegrants, dispersants, solubilizers, dyes, emulsifiers, emulsion stabilizers, fillers, film-forming agents, flavor enhancers, flavoring agents, flow enhancers, gelling agents, granulators, humectants, lubricants, mucosal adhesives, ointment bases, ointments, oily mediators, organic bases, lozenge bases, pigments, plasticizers, polishing agents, preservatives, multivalent chelating agents, skin penetrants, solubilizers, solvents, stabilizers, suppository bases, and surface active agents. Agents, surfactants, suspending agents, sweeteners, therapeutic agents, thickeners, tension agents, toxic agents, viscous agents, water-miscible cosolvents, water softeners, or wetting agents. See, for example, Handbook of Pharmaceutical Excipients, 3rd edition, AH Kibbe (Pharmaceutical Press, London, 2000) (included in its entirety by reference); Remington's Pharmaceutical Sciences, 16th edition, EW Martin (Mack Publishing Co., Easton, Pennsylvania, 1980) (included in its entirety by reference).

[0488] In an exemplary aspect, the pharmaceutical composition comprises a formulation material that is non-toxic to the recipient at the dosage and concentration used. In a particular embodiment, the pharmaceutical composition comprises an active pharmaceutical agent and one or more pharmaceutically acceptable salts; polyols; surfactants; osmotic balancers; toning agents; antioxidants; antibiotics; antifungal agents; swelling agents; lyophilization protectants; antifoaming agents; chelating agents; preservatives; colorants; analgesics; or other pharmaceutical agents. In an exemplary aspect, optionally in addition to one or more excipients, the pharmaceutical composition also comprises one or more polyols and / or one or more surfactants, wherein the one or more excipients include, but are not limited to, pharmaceutically acceptable salts; osmotic balancers (toning agents); antioxidants; antibiotics; antifungal agents; swelling agents; lyophilization protectants; antifoaming agents; chelating agents; preservatives; colorants; and analgesics.

[0489] In some embodiments, the pharmaceutical composition may contain formulation materials for altering, maintaining, or preserving, for example, the composition's pH, osmotic pressure, viscosity, transparency, color, isotonicity, odor, sterility, stability, dissolution or release rate, adsorption, or permeation. In such embodiments, suitable formulation materials include, but are not limited to, amino acids (such as glycine, glutamine, asparagine, arginine, or lysine); antimicrobial agents; antioxidants (such as ascorbic acid, sodium sulfite, or sodium bisulfite); buffers (such as borates, bicarbonates, Tris-HCl, citrates, phosphates, or other organic acids); swelling agents (such as mannitol or glycine); chelating agents (such as ethylenediaminetetraacetic acid (EDTA)); complexing agents (such as caffeine, polyvinylpyrrolidone, β-cyclodextrin, or hydroxypropyl-β-cyclodextrin); fillers; monosaccharides; disaccharides; and other carbohydrates (such as glucose, mannose, or dextrin); proteins (such as serum albumin, gelatin, or immunoglobulins); colorants, flavorings, and diluents; emulsifiers; hydrophilic polymers (such as polyvinylpyrrolidone); low molecular weight peptides; and salts. Counterions (e.g., sodium); preservatives (e.g., benzalkonium chloride, benzoic acid, salicylic acid, thimerosal, phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbic acid, or hydrogen peroxide); solvents (e.g., glycerol, propylene glycol, or polyethylene glycol); sugar alcohols (e.g., mannitol or sorbitol); suspending agents; surfactants or wetting agents (e.g., pluronics, PEG, dehydrated sorbitol esters, polysorbate esters (e.g., polysorbate 20, polysorbate esters), triton, tromethamine, lecithin, cholesterol, tyloxacin); stability enhancers (e.g., sucrose or sorbitol); tension enhancers (e.g., alkali metal halides (preferably sodium chloride or potassium chloride), mannitol, sorbitol); delivery media; diluents; excipients and / or pharmaceutical adjuvants. See Remington's Pharmaceutical Sciences, 18th edition (edited by AR Genrmo), 1990, Mack Publishing Company.

[0490] The pharmaceutical composition may be formulated to achieve a physiologically compatible pH. In some embodiments, the pH of the pharmaceutical composition may be, for example, between about 4 or about 5 and about 8.0, or between about 4.5 and about 7.5, or between about 5.0 and about 7.5. In an exemplary embodiment, the pH of the pharmaceutical composition is between 5.5 and 7.5.

[0491] 4.2 Treatment methods This disclosure also provides treatment methods. In an exemplary embodiment, the method is a way of treating a subject in need, which includes administering to the subject in need an amount of the pharmaceutical composition of this disclosure that is effective in treating the subject.

[0492] This document provides methods for treating subjects with cancer and methods for treating subjects with solid tumors. In an exemplary embodiment, the method includes administering to the subject an amount of the pharmaceutical composition disclosed herein that is effective in treating the subject's cancer or solid tumor.

[0493] The cancers that can be treated by the methods disclosed herein can be any cancer, such as any malignant growth or tumor caused by abnormal and uncontrolled cell division that can spread to other parts of the body via the lymphatic system or bloodstream. In some respects, cancer is selected from the following groups: acute lymphoblastic carcinoma, acute myeloid leukemia, alveolar rhabdomyosarcoma, bone cancer, brain cancer, breast cancer, cancer of the anus, anal canal or anorectal region, eye cancer, intrahepatic bile duct cancer, joint cancer, cancer of the neck, gallbladder or pleura, cancer of the nose, nasal cavity or middle ear, oral cancer, vulvar cancer, chronic lymphocytic leukemia, chronic myeloid carcinoma, colon cancer, esophageal cancer, cervical cancer, gastrointestinal carcinoid tumors, Hodgkin's lymphoma, hypopharyngeal cancer, kidney cancer, laryngeal cancer, liver cancer, lung cancer, malignant mesothelioma, melanoma, multiple myeloma, nasopharyngeal carcinoma, non-Hodgkin's lymphoma, ovarian cancer, pancreatic cancer, cancer of the peritoneum, omentum and mesentery, pharyngeal cancer, prostate cancer, rectal cancer, kidney cancer (e.g., renal cell carcinoma (RCC)), small bowel cancer, soft tissue cancer, stomach cancer, testicular cancer, thyroid cancer, ureteral cancer and bladder cancer. In certain aspects, the cancer is selected from the group consisting of: head and neck cancer, ovarian cancer, cervical cancer, bladder cancer and esophageal cancer, pancreatic cancer, gastrointestinal cancer, gastric cancer, breast cancer, endometrial cancer and colorectal cancer, hepatocellular carcinoma, glioblastoma, bladder cancer, lung cancer (e.g., non-small cell lung cancer (NSCLC)), and bronchioloalveolar carcinoma. In certain embodiments, the tumor is non-small cell lung cancer (NSCLC), head and neck cancer, kidney cancer, triple-negative breast cancer, and gastric cancer. In an exemplary aspect, a subject has a tumor (e.g., a solid tumor, a hematologic malignancy, or a lymphoma) and the subject is administered an amount of pharmaceutical composition effective in treating the subject's tumor. In other exemplary aspects, the tumor is non-small cell lung cancer (NSCLC), small cell lung cancer (SCLC), head and neck cancer, kidney cancer, breast cancer, melanoma, ovarian cancer, liver cancer, pancreatic cancer, colon cancer, prostate cancer, gastric cancer, lymphoma, or leukemia, and the subject is administered an amount of pharmaceutical composition effective in treating the subject's tumor.

[0494] As used herein, the term "treatment" and related terms do not necessarily imply 100% or complete cure. Rather, there are different degrees of treatment that a person skilled in the art would consider to have potential benefit or therapeutic effect. In this regard, the methods for treating cancer disclosed herein can provide any amount or level of treatment. Furthermore, treatment provided by the methods disclosed herein can include treating one or more symptoms or signs of the cancer being treated. Treatment provided by the methods disclosed herein can also cover slowing the progression of cancer. For example, these methods can treat cancer by enhancing T-cell activity or the immune response against cancer, reducing tumor or cancer growth, reducing tumor cell metastasis, increasing cell death of tumor or cancer cells, etc. In an exemplary aspect, these methods provide treatment by delaying the onset or recurrence of cancer by 1 day, 2 days, 4 days, 6 days, 8 days, 10 days, 15 days, 30 days, two months, 4 months, 6 months, 1 year, 2 years, 4 years, or longer. In an exemplary aspect, these methods provide treatment by increasing the survival rate of the subject.

[0495] Specifically, the antigen-binding protein disclosed in this paper targets cancer-associated fibroblasts present in the tumor stroma. The tumor stroma (broadly defined as the non-cancerous and non-immune cell component of a tumor) has traditionally been considered a structural component that holds tumor tissue together. The tumor stroma consists of extracellular matrix and specialized connective tissue cells, including fibroblasts and mesenchymal stromal cells. Tumors typically require the stroma for nutrition and waste removal, but its content can vary significantly across different types of cancer. For example, many lymphomas have very little stroma, while the stroma can constitute up to 90% of other solid tumors. The antigen-binding protein disclosed in this paper specifically targets FAP+ tumors. Fibroblasts can infiltrate tumors, and FAP+ cells can be readily identified using methods well-known in the art, such as immunostaining.

[0496] 4.3 Subjects In some embodiments disclosed herein, the subject is a mammal, including but not limited to rodent mammals such as mice and hamsters; and lagomorph mammals such as rabbits; mammals from the order Carnivora, including felines (cats) and canines (dogs); mammals from the order Artiodactyla, including bovids (cows) and suidae (pigs); or perissodactyl mammals, including equines (horses). In some aspects, the mammal belongs to the order Primates, Ceboid, or Simoid (monkeys) or Anthropoids (humans and apes). In some aspects, the mammal is human.

[0497] 5. Reagent kit This disclosure further provides kits comprising the antigen-binding proteins, nucleic acids, vectors, or host cells or combinations thereof disclosed herein. In an exemplary aspect, the antigen-binding proteins, nucleic acids, vectors, or host cells are provided in the kit in unit doses. For purposes herein, "unit dose" refers to a discrete amount dispersed in a suitable carrier. In an exemplary aspect, a unit dose is an amount sufficient to provide a desired effect (e.g., cancer treatment) to a subject. In an exemplary aspect, the kit comprises several unit doses, for example, a weekly or monthly supply of unit doses, optionally packaged individually or otherwise separately from other unit doses. In some embodiments, components / unit doses of the kit are packaged together with instructions for administration to a patient. In some embodiments, the kit comprises one or more devices for administration to a patient, such as needles and syringes. In some aspects, the antigen-binding proteins, nucleic acids, vectors, host cells, or combinations thereof are pre-packaged in ready-to-use forms (e.g., syringes, intravenous infusion bags, etc.). In an exemplary aspect, the ready-to-use form is for single-use only. In an exemplary aspect, the kit comprises multiple single-use ready-to-use forms of the antigen-binding proteins, nucleic acids, vectors, or host cells disclosed herein. In some respects, the kit further includes other therapeutic or diagnostic agents or pharmaceutically acceptable carriers (e.g., solvents, buffers, diluents, etc.), including any of the therapeutic or diagnostic agents or pharmaceutically acceptable carriers described herein.

[0498] The following examples are given only to illustrate the invention and are not intended to limit its scope in any way.

[0499] Example Example 1. Generation and characterization of anti-FAP antibodies 1. Anti-FAP antibody production Fully human antibodies against human FAP were generated by immunizing XENOMOUSE® transgenic mice (US Patent Nos. 6,114,598; 6,162,963; 6,833,268; 7,049,426; 7,064,244, which are incorporated herein by reference in their entirety; Green et al., 1994, Nature Genetics 7:13-21; Mendez et al., 1997, Nature Genetics 15:146-156; Green and Jakobovits, 1998, J. Ex. Med, 188:483-495; Kellerman and Green, Current Opinion in Biotechnology 13, 593-597, 2002). Animals from the XMG4 and XMG2 XENOMOUSE® strains were used for these immunizations. Multiple immunogens and immunization pathways were used to generate an anti-human FAP immune response. For gene immunization, mice were immunized 11 times over 4–5 weeks using the Helios gene gun system, according to the manufacturer's instructions (BioRad). Briefly, an expression vector encoding wild-type human FAP was coated onto a gold bead (BioRad) and delivered to the epidermis of the abdomen of shaved mice. For soluble protein immunization, mice were immunized with recombinant human FAP protein representing the extracellular domain. Animals were immunized 10 times over 4–5 weeks using subcutaneous injections of the recombinant protein mixed with Alum and CpG-ODN. The initial injection was 10 µg, with subsequent booster doses of 5 µg. FAP-specific serum titers were monitored using transiently transfected 293T cells via live-cell FACS analysis on an Accuri flow cytometer (BD Biosciences). Animals exhibiting the highest antigen-specific serum titers against human FAP were sacrificed and used to generate hybridomas (Kohler and Milstein, 1975). Pooled lymphocytes from the spleen and / or draining lymph nodes (from each harvest) were isolated from lymphoid tissue by grinding in a suitable medium (e.g., Durbeco Modified Eagle Medium (DMEM); Invitrogen, Carlsbad, CA). B cells were selected and / or expanded using standard methods and fused with suitable fusion couples using techniques known in the art. Hybridoma supernatants conjugated with both human and cynomolgus monkey FAP were then selected for further characterization. Table 1 summarizes the number of identified antigen-specific hits.

[0500] Table 1. Number of antigen-specific antibodies against human FAP protein and cynomolgus monkey FAP protein identified from Xenomouse activities 2. Additional binding characterization of anti-FAP antibodies Depleted hybridoma supernatants were tested by flow cytometry for binding to human FAP and cynomolgus FAP. To confirm binding to human FAP and cynomolgus FAP, as well as binding lacking human DPP4, HEK293T cells were transiently transfected with mammalian expression constructs encoding human FAP, cynomolgus FAP, mouse FAP, or human DPP4 using 293Fectin (Thermo Fisher). To confirm antibody binding to endogenously expressed human FAP, depleted hybridoma supernatants were also tested using RPMI-7951 cells endogenously expressing human FAP. SKML-28 cells lacking endogenous FAP expression were used as a negative control. Cells were incubated with depleted hybridoma supernatant normalized to 5 μg / mL antibody, followed by incubation with Alexa Fluor 647-conjugated goat anti-human IgG secondary antibody (Jackson Immuno Research). Dead cells were stained with 7-aminoactinomycin D (Sigma). Cells were run on an Accuri flow cytometer to detect anti-FAP antibody binding. Generally, there was a good correlation between human FAP and cynomolgus monkey FAP binding, as well as between endogenous human FAP and transiently expressed human FAP binding. Cross-reactivity was found between single antibodies and mouse FAP, while no cross-reactivity with human DPP4 was observed for any of the antibodies tested. Table 2-4 summarizes the binding characterization of these antibodies.

[0501] Table 2. Binding characterization of anti-FAP antibodies Table 3. Binding to 293T cells transiently expressing human FAPα, cyno FAPα, mouse FAPα, and human DPP4 (FACS geometric mean) Table 4. Binding to endogenously expressed human FAPα (FACS geometric mean) 3. Epitope grouping analysis of anti-FAP antibodies A common method for characterizing epitopes is through competition assays. Competing antibodies can be considered as binding to the same or overlapping sites on the target. This example describes a method for identifying competitive binding sites of human FAP, and the results of this method when applied to many of the antibodies described herein.

[0502] Grouping experiments can be performed in various ways, and the methods used can affect the assay results. What these methods have in common is that FAP is typically bound by one reference antibody and detected by another. If the reference antibody prevents the binding of the probe antibody, these antibodies are considered to be in the same group. The order in which the antibodies are used is important. If antibody A is used as the reference antibody and blocks the binding of antibody B, the reverse is not always true: antibody B, used as the reference antibody, does not necessarily block antibody A. Several factors come into play here: antibody binding can cause a conformational change in the target, thus preventing the binding of the second antibody, or overlapping but not completely masking epitopes can allow the second antibody to still interact with the target with a sufficiently high affinity to allow binding. Generally, if competition is observed in either order, the antibodies are said to be grouped together, and if two antibodies can block each other, it is likely that the epitopes overlap more completely.

[0503] In this example, a modified antibody-antibody competition assay was used to determine the relative epitope grouping profiles of FAP-specific antibodies in a high-throughput manner. In short, the ability of each antibody to competitively bind to a group of reference antibodies selected based on their distinct binding characteristics according to approximate groupings determined from preliminary data generated using smaller sample sets representing different yields was tested. The competition / binding pattern of each test antibody against the reference antibody group was then determined and compared to that generated from other test antibodies. The correlation between the competition / binding profiles of the individual test antibodies was then compared. Antibodies exhibiting similar competition / binding profiles were grouped (classified) together (e.g., grouping profiles A, B, etc.).

[0504] Biotinylated recombinant soluble human FAP protein was conjugated with streptoacidin-coated, uniquely barcoded LumAvidin Beads® (Luminex) in the dark for 45 minutes at room temperature, followed by two washes. The reference antibody hybridoma supernatant sample was incubated with the antigen-coated beads in the dark for 1 hour at room temperature, followed by three washes. The beads were resuspended in FACS buffer containing Stabilguard® (SurModics). The antigen-coated reference antibody binding beads were pooled and then aliquoted into wells containing normalized (5 µg / ml) test antibody (hybridoma supernatant) samples (or negative controls), incubated in the dark for 1 hour at room temperature, followed by two washes. The samples were then incubated with Alexa Fluor® 488 IgG fragment-specific detection antibody (Jackson Immunological Research) in the dark for 15 minutes at room temperature, followed by one wash and resuspended in FACS buffer. Samples were analyzed using an Accuri flow cytometer equipped with an Intellicyt HyperCyt autosampler.

[0505] To determine the antibody competition / binding profiles for each test antibody, for each competition / binding reaction (i.e., the entire reference antibody set), the reference antibody binding signal alone was subtracted from the reference antibody plus the test antibody signal. Each antibody binding profile was defined as the set of net binding values ​​for each competition / binding reaction. The similarity between profiles was then assessed by calculating the coefficient of determination between each test antibody profile. Test antibodies showing high similarity to each other (R² > 0.8) were then grouped into a common group profile. Individual group profiles were defined only when two or more highly correlated samples were present. If a single unique antibody group profile was observed (i.e., they showed a low degree of similarity to other test antibody binding profiles), the group was classified as unknown. Of the 554 samples tested, 499 were subdivided into 14 unique group profiles. Table 5 summarizes the grouping data showing the number of samples in each group across three harvests.

[0506] Table 5. Antibody grouping profile of anti-FAP antibodies 4. Molecular rescue and sequencing of anti-FAP antibodies RNA (total RNA or mRNA) was purified from wells containing hybridoma cells producing anti-FAP antibodies using the Qiagen RNeasy Mini Kit or the Invitrogen mRNA catcher plus kit. The purified RNA was used to amplify the antibody heavy and light chain variable region (V) genes using cDNA synthesis via reverse transcription followed by polymerase chain reaction (RT-PCR). The fully human antibody γ heavy chain was obtained using the Qiagen One-Step Reverse Transcription PCR Kit (Qiagen). This method was used to generate first-strand cDNA from an RNA template, followed by multiplex PCR amplification of the γ heavy chain variable region. The 5' γ chain-specific primer was annealed to the antibody heavy chain signal sequence, while the 3' primer was annealed to the γ constant domain region. The fully human κ light chain was obtained using the Qiagen One-Step Reverse Transcription PCR Kit (Qiagen). This method was used to generate first-strand cDNA from an RNA template, followed by multiplex PCR amplification of the κ light chain variable region. The 5' κ light chain-specific primer was annealed to the signal sequence of the antibody light chain, while the 3' primer was annealed to the region of the κ constant domain. The complete human λ light chain was obtained using the Qiagen One-Step Reverse Transcription PCR Kit (Qiagen). This method is used to generate first-strand cDNA from an RNA template, followed by multiplex PCR amplification of the variable region of the λ light chain. The 5' λ light chain-specific primer was annealed to the signal sequence of the light chain, while the 3' primer was annealed to the region of the λ constant domain.

[0507] The amplified cDNA was enzymatically purified using exonuclease I and alkaline phosphatase, and the purified PCR products were directly sequenced. Amino acid sequences were deduced from the corresponding nucleic acid sequences using bioinformatics methods. Each hybridoma sample underwent two additional independent RT-PCR amplification and sequencing cycles to confirm that any observed mutations were not the result of PCR. The resulting amino acid sequences were then analyzed to determine the germline sequence origin of the antibody and to identify deviations from the germline sequence. Comparisons of each heavy and light chain sequence with its original germline sequence were indicated. Amino acid sequences corresponding to the complementarity-determining regions (CDRs) of the sequenced antibody were aligned, and clones were grouped by similarity using these alignments.

[0508] Using the method described above, 82 unique heavy chains from 41 different VDJ recombinants were resolved. Among these antibodies, the light chain sequences of two antibodies from each VDJ group were resolved, resulting in the identification of 37 unique antibodies with sequence resolution from 29 different VDJ groups. Of these, 24 unique antibodies with 24 different CDR3 sequences from 24 different VDJ groups were selected for the generation of bispecific molecules (Table 6). Groups C and D are subgroups and can be considered substantially identical.

[0509] Table 6. Anti-FAP antibodies Example 2. Generation and characterization of bispecific molecules targeting FAP This example describes the generation and...

Claims

1. A fibroblast activating protein α (FAP) antigen-binding protein, the FAP antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein comprises: (1) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 197, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 198; (2) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 199, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 200; (3) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 201, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 202; (4) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 203, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 204; (5) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 205, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 206; (6) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 207, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 208; (7) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 209, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 210; (8) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 211, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 212; (9) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 213, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 214; (10) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 215, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 216; (11) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 217, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 218; (12) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 219, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 220; (13) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 221, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 222; (14) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 223, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 224; (15) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 225, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 226; (16) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 227, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 228; (17) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 229, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 230; (18) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 231, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 232; (19) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 233, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 234; (20) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 235, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 236; (21) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 237, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 238; (22) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 239, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 240; (23) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 241, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 242; (24) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 243, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 244; (25) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 245, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 246; (26) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 247, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 248; (27) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 249, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 250; (28) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 251, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 252; (29) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 253, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 254; (30) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 255, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 256; (31) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 257, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 258; (32) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 259, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 260; (33) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 261, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 262; (34) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 263, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 264; (35) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 265, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 266; (36) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 267, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 268; (37) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 269, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 270; (38) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 271, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 272; (39) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 273, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 274; (40) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 275, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 276; (41) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 277, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 278; (42) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 279, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 280; (43) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 281, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 282; (44) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 283, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 284; (45) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 285, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 286; (46) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 287, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 288; (47) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 289, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 290; (48) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 291, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 292; (49) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 293, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 294; (50) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 295, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 296; (51) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 297, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 298; (52) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 299, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 300; (53) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 301, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 302; (54) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 303, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 304; (55) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 305, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 306; (56) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 307, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 308; (57) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 309, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 310; (58) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 311, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 312; (59) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 313, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 314; (60) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 315, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 316; (61) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 317, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 318; (62) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 319, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 320; (63) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 321, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 322; (64) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 323, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 324; (65) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 325, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 326; (66) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 327, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 328; (67) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 329, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 330; (68) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 331, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 332; (69) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 333, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 334; (70) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 335, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 336; (71) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 337, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 338; (72) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 339, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 340; or (73) The heavy chains CDR-H1, CDR-H2 and CDR-H3 of SEQ ID NO: 341, and the light chains CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO:

342.

2. The FAP antigen-binding protein of claim 1, wherein the FAP antigen-binding protein comprises: (1) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 1-6 respectively; (2) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 6-12 respectively; (3) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 13-18; (4) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 19-24 respectively; (5) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 25-30; (6) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 31-36 respectively; (7) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 37-42; (8) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 43-48; (9) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 49-54; (10) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 55-60 respectively; (11) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 61-66; (12) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 67-72 respectively; (13) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 73-78; (14) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 79-84; (15) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 85-90; (16) Each of the following contains CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 of SEQ ID NO: 91-96 respectively; (17) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 97-102; (18) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 103-108; (19) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 109-114; (20) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 115-120; (21) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 121-126; (22) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 127-132; (23) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 133-138; (24) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 139-144; (25) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 145-150; (26) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 151-156; (27) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 157-162; (28) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 163-168; (29) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 169-174; (30) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 175-180; (31) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 181-186; (32) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 187-192; (33) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 43, 44, 45, 46, 47 and 193; (34) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 43, 44, 194, 46, 47 and 193; (35) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 175, 176, 177, 178, 179 and 195; (36) CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and CDR-L3 respectively containing SEQ ID NO: 187, 188, 196, 190, 191 and 192; (37) CDR-H1 containing any one of SEQ ID NO: 398-402; CDR-H2 containing any one of SEQ ID NO: 403-407; CDR-H3 containing any one of SEQ ID NO: 408-412; CDR-L1 containing any one of SEQ ID NO: 413-417; CDR-L2 containing any one of SEQ ID NO: 418-422; and CDR-L3 containing any one of SEQ ID NO: 423-427; (38) CDR-H1 containing any one of SEQ ID NO: 428-432; CDR-H2 containing any one of SEQ ID NO: 433-437; CDR-H3 containing any one of SEQ ID NO: 438-442; CDR-L1 containing any one of SEQ ID NO: 443-447; CDR-L2 containing any one of SEQ ID NO: 448-452; and CDR-L3 containing any one of SEQ ID NO: 453-457; (39) A CDR-H1 containing any one of SEQ ID NO: 458-462; a CDR-H2 containing any one of SEQ ID NO: 463-467; a CDR-H3 containing any one of SEQ ID NO: 468-472; a CDR-L1 containing any one of SEQ ID NO: 473-477; a CDR-L2 containing any one of SEQ ID NO: 478-482; and a CDR-L3 containing any one of SEQ ID NO: 483-487; or (40) A CDR-H1 comprising any one of SEQ ID NO: 488-492; a CDR-H2 comprising any one of SEQ ID NO: 493-497; a CDR-H3 comprising any one of SEQ ID NO: 498-502; a CDR-L1 comprising any one of SEQ ID NO: 503-507; a CDR-L2 comprising any one of SEQ ID NO: 508-512; and a CDR-L3 comprising any one of SEQ ID NO: 513-517.

3. A fibroblast activator protein α (FAP) antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein binds to an epitope comprising residues V275, R175, F181, Q182, I183, D178, F185, P179 and Y274 as numbered according to SEQ ID NO:

394.

4. The FAP antigen-binding protein of claim 3, wherein the VH and the VL comprise (VH and VL numbered according to Kabat): (1) H33 is Arg, Lys, Gln or Asn; (2) H94 is Arg, Lys, Gln or Asn; (3) H97 is Gly or Ala; (4) H98 is Tyr, Trp, Phe, Thr or Ser; (5) H100B is Tyr, Trp, Phe, Thr or Ser; (6) H100C is Tyr, Trp, Phe, Thr or Ser; (7) L53 is Gln, Asn or Glu; (8) L54 is Arg, Lys, Gln or Asn; and (9) L60 is Asp, Glu or Asn.

5. The FAP antigen-binding protein as described in claim 3 or 4, wherein: (a) The VH comprises: (i) a complementary determination region (CDR)-H1 comprising any one of SEQ ID NO: 398-402; and (ii) a CDR-H3 comprising any one of SEQ ID NO: 408-412; (b) The VL comprises a CDR-L2 containing any one of SEQ ID NO: 418-422; and (c) The VL is further included in the D at position L60 (according to the Kabat number).

6. A fibroblast activating protein α (FAP) antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein binds to epitopes comprising E325, D331, Q336, I320, R324, E302, R303 and T335 according to SEQ ID NO:

394.

7. The FAP antigen-binding protein of claim 6, wherein the VH comprises (VH numbered according to Kabat): (1) H31 is Arg, Lys, Gln or Asn; (2) H98 is Tyr, Trp, Phe, Thr or Ser; (3) H99 is Tyr, Trp, Phe, Thr or Ser; (4) H100 is Tyr, Trp, Phe, Thr or Ser; and (5) H101 is Asp, Glu or Asn.

8. The FAP antigen-binding protein as described in claim 6 or 7, wherein: (a) The VH comprises: (i) a complementary determination region (CDR)-H1 comprising any one of SEQ ID NO: 428-432; and (ii) a CDR-H3 comprising any one of SEQ ID NO: 438-442; and (b) The VL comprises a CDR-L2 containing any one of SEQ ID NO: 448-452.

9. A fibroblast activator protein α (FAP) antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein binds to epitopes comprising S86, E82, I181, T83, Q85, Q65, K486, N80, T88, I485 and I487 according to SEQ ID NO:

394.

10. The FAP antigen-binding protein of claim 9, wherein the VH comprises (VH numbered according to Kabat): (1) H31 is Asn, Gln, His, Asp, Lys, or Arg; (2) H33 is Gly or Ala; (3) H52A is Tyr, Trp, Phe, Thr, or Ser; (4) H55 is Arg, Lys, Gln, or Asn; (5) H56 is Asn, Gln, His, Asp, Lys, or Arg; (6) H95 is Asp, Glu, or Asn; (7) H100 is Gly or Ala; (8) L32 is Tyr, Trp, Phe, Thr, or Ser; (9) L91 is Phe, Leu, Val, Ile, Ala, or Tyr; and (10) L95 is Tyr, Trp, Phe, Thr, or Ser.

11. The FAP antigen-binding protein as described in claim 9 or 10, wherein: (a) The VH comprises: (i) a CDR-H1 comprising any one of SEQ ID NO: 458-462; (ii) a CDR-H3 comprising any one of SEQ ID NO: 463-467; and (iii) a CDR-H3 comprising any one of SEQ ID NO: 468-472; and (b) The VL comprises: (i) a CDR-L1 comprising any one of SEQ ID NO: 473-487; (ii) a CDR-L2 comprising any one of SEQ ID NO: 478-482; and (iii) a CDR-L3 comprising any one of SEQ ID NO: 483-487.

12. A fibroblast activator protein α (FAP) antigen-binding protein comprising a heavy chain variable domain (VH) and a light chain variable domain (VL), wherein the protein binds to epitopes comprising K381, K371, E414, S428, Q389, Y432, I390, P434 and K436 according to SEQ ID NO:

394.

13. The FAP antigen-binding protein of claim 12, wherein the VH and the VL comprise (VH and VL numbered according to Kabat): (1) H28 is Thr or Ser; (2) H53 is Asp, Glu or Asn; (3) H96 is Arg, Lys, Gln or Asn; (4) H99 is Tyr, Trp, Phe, Thr or Ser; (5) H100A is Tyr, Trp, Phe, Thr or Ser; (6) H100B is Tyr, Trp, Phe, Thr or Ser; and (7) H100C is Tyr, Trp, Phe, Thr or Ser.

14. The FAP antigen-binding protein as described in claim 12 or 13, wherein: (a) The VH comprises: (i) a CDR-H1 comprising any one of SEQ ID NO: 488-492; (ii) a CDR-H2 comprising any one of SEQ ID NO: 493-497; and (iii) a CDR-H3 comprising any one of SEQ ID NO: 498-502; and (b) The VL comprises: (i) a CDR-L1 comprising any one of SEQ ID NO: 503-507; and (ii) a CDR-L3 comprising any one of SEQ ID NO: 513-517.

15. The FAP antigen-binding protein according to any one of claims 1-14, wherein the FAP antigen-binding protein comprises: a VL frame derived from a human Vκ frame sequence or a human Vλ frame sequence, and a VH frame derived from a human VH1, VH2, VH3, VH4 or VH5 frame sequence.

16. The FAP antigen-binding protein according to any one of claims 1-15, wherein the FAP antigen-binding protein is IgG.

17. The FAP antigen-binding protein according to any one of claims 1-15, wherein the FAP antigen-binding protein is an antigen-binding fragment of IgG.

18. The FAP antigen-binding protein according to any one of claims 1-15, wherein the FAP antigen-binding protein is a single-chain Fv (scFv).

19. The FAP antigen-binding protein of claim 18, wherein the scFv comprises a first linker between VH and VL.

20. A nucleic acid comprising a nucleotide sequence encoding the FAP antigen-binding protein as described in any one of claims 1-19.

21. A host cell comprising the nucleic acid as described in claim 20.

22. A pharmaceutical composition comprising (i) the FAP antigen-binding protein as described in any one of claims 1-19; and (ii) a pharmaceutically acceptable carrier, excipient, or diluent.

23. A method for preparing the FAP antigen-binding protein as described in any one of claims 1-19 under conditions of expressing the FAP antigen-binding protein.

24. A method of treating cancer, the method comprising administering to a subject in need a therapeutically effective amount of the FAP antigen-binding protein as described in any one of claims 1-19 or the pharmaceutical composition as described in claim 22.

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