Nova Patents
IL280618A

Proteins binding nkg2d, cd16 and a tumor-associated antigen

Abstract

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IL280618A, drawing sheet 1
Sheet 1 of 23

Term

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12 claims: 10 independent, 2 dependent

  1. 1
    WHAT IS CLAIMED IS:1. A protein comprising: (a) a first antigen-binding site that binds NKG2D;(b) a second antigen-binding site that binds a tumor-associated antigen cMET;and (c) an antibody Fc domain or a portion thereof sufficient to bind CD 16, or a third antigen-binding site that binds CD 16. 2. A protein comprising: (a) a first antigen-binding site that binds NKG2D;(b) a second antigen-binding site that binds a tumor-associated antigen selected from the group consisting of KIT, F3, IGF1R, Lewis Y, MUC13, MUC4, MCAM, LRRC32, sialyl-Tn, gpA33, GD3, GM2, c-MET, EPHA3, TNFRSF10A, TNFSF11, CD74, and PMEL;and (c) an antibody Fc domain or a portion thereof sufficient to bind CD 16, or a third antigen-binding site that binds CD 16. 3. The protein of claim 1 or 2, further comprising an additional antigen-binding site that binds the same tumor-associated antigen as the second tumor-associated antigenbinding site. 4. The protein of any one of claims 1-3, wherein the first antigen-binding site that binds NKG2D is a single-chain variable fragment (scFv), and the second and/or the additional antigen-binding site that binds a tumor-associated antigen is an Fab fragment. 5. The protein of any one of claims 1-3, wherein the first anti gen-binding site that binds NKG2D is an scFv, and the second and/or the additional antigen-binding site that binds a tumor-associated antigen is an scFv. 6. The protein of claim 1 or 2, wherein the first antigen-binding site that binds NKG2D is an Fab fragment, and the second antigen-binding site that binds a tumorassociated antigen is an scFv. 7. The protein of claim 1 or 2, wherein the first antigen-binding site that binds NKG2D is an scFv, and the second antigen-binding site that binds a tumor-associated antigen is an Fab fragment. 8. The protein of any one of claims 1-7, wherein the first anti gen-binding site binds to NKG2D in humans. 9. The protein of any one of claims 1-8, wherein the first, the second, and/or the additional antigen-binding site comprises a heavy chain variable domain and a light chain variable domain. 10. The protein of any one of claims 4-7, wherein the scFv that binds the tumorassociated antigen and/or the scFv that binds NKG2D is linked to an antibody constant domain or a portion thereof sufficient to bind CD 16, via a hinge comprising Ala-Ser, wherein the scFv comprises a heavy chain variable domain and a light chain variable domain. 11. The protein according to any one of claims 9-10, wherein the heavy chain variable domain forms a disulfide bridge with the light chain variable domain. 12. The protein according to claim 11, wherein the disulfide bridge is formed between C44 from the heavy chain variable domain and Cl00 from the light chain variable domain. 13. The protein according to any one of claims 10-12, wherein the scFv the heavy chain variable domain is linked to the light chain variable domain via a flexible linker. 14. The protein according to claim 13, wherein within the scFv the flexible linker comprises (G4S)4. 15. The protein according to any one of claims 10-14, wherein within the scFv the heavy chain variable domain is positioned at the N-terminus or the C-terminus of the light chain variable domain. 16. The protein according to any one of claims 10-15, wherein within the scFv the hinge further comprises amino acid sequence Thr-Lys-Gly. 17. The protein according any one of claims 1-16, wherein the first antigenbinding site that binds NKG2D comprises: (1) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 387, 88, and 390, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;
  2. 2
    (2) a heavy chain variable domain comprising complementarity-determining region 1 (CDR1), complementarity-determining region 2 (CDR2), and complementarity-determining region 3 (CDR3) sequences represented by the amino acid sequences of SEQ ID NOs:387, 88, and 412, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;
  3. 3
    (3) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:435, 64, and 436, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 66, 67, and 68, respectively;
  4. 4
    (4) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:437, 72, and 408, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 74, 75, and 76, respectively;
  5. 5
    (5) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:409, 80, and 411, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 82, 83, and 84, respectively;
  6. 6
    (6) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:421, 96, and 422, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 98, 99, and 100, respectively;
  7. 7
    (7) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:87, 88, and 89, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;
  8. 8
    (8) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:387, 88, and 393, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;
  9. 9
    (9) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:387, 88, and 396, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;
  10. 10
    (10) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:387, 88, and 399, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;
  11. 11
    (11) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:387, 88, and 402, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;
  12. 12
    (12) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs:387, 88, and 405, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;or (13) a heavy chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 87, 88, and 389, respectively;and a light chain variable domain comprising CDR1, CDR2, and CDR3 sequences represented by the amino acid sequences of SEQ ID NOs: 90, 91, and 92, respectively;18. The protein according any one of claims 1-17, wherein the first antigenbinding site that binds NKG2D comprises: (1) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:388 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:86;(2) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:85 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:86;(3) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:61 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:62;(4) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:69 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:70;(5) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:77 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:78;(6) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:93 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:94;(7) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:391 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:86;(8) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:394 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:86;(9) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:397 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:86;(10) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:400 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:86;or (11) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:403 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:86. 19. The protein according any one of claims 1-16, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to an amino acid sequence selected from: SEQ ID NO:1, SEQ ID NO:41, SEQ ID NO:49, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:69, SEQ ID NO:77, SEQ ID NO:85, and SEQ ID NO:93. 20. The protein according any one of claims 1-16, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:41 and a light chain variable domain at least 90% identical to SEQ IDNO:42. 21. The protein according any one of claims 1-16, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:49 and a light chain variable domain at least 90% identical to SEQ IDNO:50. 22. The protein according any one of claims 1-16, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:57 and a light chain variable domain at least 90% identical to SEQ IDNO:58. 23. The protein according any one of claims 1-16, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:59 and a light chain variable domain at least 90% identical to SEQ IDNO:60. 24. The protein according any one of claims 1-18, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:61 and a light chain variable domain at least 90% identical to SEQ IDNO:62. 25. The protein according any one of claims 1-18, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:69 and a light chain variable domain at least 90% identical to SEQ IDNO:70. 26. The protein according any one of claims 1-18, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:77 and a light chain variable domain at least 90% identical to SEQ IDNO:78. 27. The protein according any one of claims 1-18, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:85 and a light chain variable domain at least 90% identical to SEQ IDNO:86. 28. The protein according any one of claims 1-18, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:93 and a light chain variable domain at least 90% identical to SEQ IDNO:94. 29. The protein according any one of claims 1-16, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 101 and a light chain variable domain at least 90% identical to SEQ ID NO: 102. 30. The protein according any one of claims 1-16, wherein the first antigenbinding site that binds NKG2D comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 103 and a light chain variable domain at least 90% identical to SEQ ID NO: 104. 31. The protein according any one of claims 1-30, wherein the second antigenbinding site that binds c-MET comprises a heavy chain variable domain comprising heavy chain CDR1 (CDRH1), heavy chain CDR2 (CDRH2), and heavy chain CDR3 (CDRH3), and a light chain variable domain comprising light chain CDR1 (CDRL1), light chain CDR2 (CDRL2), and light chain CDR3 (CDRL3), wherein the amino acid sequences of CDRH1, CDRH2, CDRH3, CDRL1, CDRL2, and CDRL3 are set forth in SEQ ID NOs: 292, 406, 294, 296, 407, and 298;300, 301, 302, 410, 305, and 306;or 414, 415, 416, 418, 419, and 420, respectively. 32. The protein according any one of claims 1-31, wherein the second antigenbinding site that binds c-MET comprises: (a) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:291 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:295;(b) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:299 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:303;or (c) a heavy chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:413 and a light chain variable domain comprising an amino acid sequence at least 90% identical to SEQ ID NO:417. 33. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen KIT, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 109 and a light chain variable domain at least 90% identical to SEQ ID NO: 113. 34. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen KIT, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 117 and a light chain variable domain at least 90% identical to SEQ ID NO: 121. 35. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen F3, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 126 and a light chain variable domain at least 90% identical to SEQ ID NO: 130. 36. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen F3, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 140 and a light chain variable domain at least 90% identical to SEQ ID NO: 144. 37. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen IGF1R, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 149 and a light chain variable domain at least 90% identical to SEQ ID NO: 153. 38. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen IGF1R, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 157 and a light chain variable domain at least 90% identical to SEQ ID NO: 161. 39. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen Lewis Y, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 166 and a light chain variable domain at least 90% identical to SEQ ID NO: 170. 40. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen Lewis Y, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 174 and a light chain variable domain at least 90% identical to SEQ ID NO: 178. 41. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen MUC13, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 182 and a light chain variable domain at least 90% identical to SEQ ID NO: 186. 42. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen MCAM, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO: 192 and a light chain variable domain at least 90% identical to SEQ ID NO: 196. 43. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen MCAM, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:200 and a light chain variable domain at least 90% identical to SEQ ID NO:204. 44. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen LRRC32, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:209 and a light chain variable domain at least 90% identical to SEQ ID NO:213. 45. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen LRRC32, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:217 and a light chain variable domain at least 90% identical to SEQ ID NO:221. 46. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen sialyl-Tn, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:226 and a light chain variable domain at least 90% identical to SEQ ID NO:230. 47. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen sialyl-Tn, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:234 and a light chain variable domain at least 90% identical to SEQ ID NO:238. 48. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen gpA33, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:242 and a light chain variable domain at least 90% identical to SEQ ID NO:246. 49. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen gpA33, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:250 and a light chain variable domain at least 90% identical to SEQ ID NO:254. 50. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen GD3, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:259 and a light chain variable domain at least 90% identical to SEQ ID NO:263. 51. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen GD3, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:267 and a light chain variable domain at least 90% identical to SEQ ID NO:271. 52. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen GM2, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:275 and a light chain variable domain at least 90% identical to SEQ ID NO:279. 53. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen GM2, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:283 and a light chain variable domain at least 90% identical to SEQ ID NO:287. 54. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen EPHA3, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:308 and a light chain variable domain at least 90% identical to SEQ ID NO:312. 55. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen TNFRSF10, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:317 and a light chain variable domain at least 90% identical to SEQ ID NO:321. 56. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen TNFRSF10, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:325 and a light chain variable domain at least 90% identical to SEQ ID NO:329. 57. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen TNFSF11, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:334 and a light chain variable domain at least 90% identical to SEQ ID NO:338. 58. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen TNFSF11, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:342 and a light chain variable domain at least 90% identical to SEQ ID NO:346. 59. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen CD74, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:351 and a light chain variable domain at least 90% identical to SEQ ID NO:355. 60. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen CD74, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:359 and a light chain variable domain at least 90% identical to SEQ ID NO:363. 61. The protein according to any one of claims 2-30, wherein the second antigen- binding site binds the tumor-associated antigen PMEL, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:369 and a light chain variable domain at least 90% identical to SEQ ID NO:373. 62. The protein according to any one of claims 2-30, wherein the second antigenbinding site binds the tumor-associated antigen PMEL, and wherein the protein comprises a heavy chain variable domain at least 90% identical to SEQ ID NO:377 and a light chain variable domain at least 90% identical to SEQ ID NO:381. 63. The protein according to any one of the preceding claims, wherein the protein comprises an antibody Fc domain or a portion thereof sufficient to bind CD16, wherein the antibody Fc domain comprises a hinge and a CH2 domain. 64. The protein according to claim 63, wherein the antibody Fc domain comprises a hinge and a CH2 domain of a human IgGl antibody. 65. The protein according to claim 63 or 64, wherein the antibody Fc domain comprises an amino acid sequence at least 90% identical to amino acids 234-332 of a human IgGl antibody. 66. The protein according to claim 63, wherein the antibody Fc domain comprises an amino acid sequence at least 90% identical to the antibody Fc domain of human IgGl and differs at one or more positions selected from the group consisting of Q347, Y349, L351, S354, E356, E357, K360, Q362, S364, T366, L368, K370, N390, K392, T394, D399, S400, D401, F405, Y407, K409, T411, and K439. 67. The protein according to any one of claims 1-18 comprising an amino acid sequence at least 95% identical to SEQ ID NO:423 and SEQ ID NO:424. 68. The protein according to claim 67 comprising an amino acid sequence at least 95% identical SEQ ID NO:425 and SEQ ID NO:426;SEQ ID NO:427 and SEQ ID NO:428;or SEQ ID NO:429 and SEQ ID NO:430. 69. A formulation comprising a protein according to any one of the preceding claims and a pharmaceutically acceptable carrier. 70. A cell comprising one or more nucleic acids expressing a protein according to any one of claims 1-68. 71. A method of enhancing tumor cell death, the method comprising exposing a tumor cell and a natural killer cell to an effective amount of the protein according to any one of claims 1-68, wherein the tumor cell expresses a tumor-associated antigen selected from the group consisting of KIT, F3, IGF1R, Lewis Y, MUC13, MUC4, MCAM, LRRC32, sialyl-Tn, gpA33, GD3, GM2, c-MET, EPHA3, TNFRSF10A, TNFSF11, CD74, and PMEL. 72. A method of treating cancer, wherein the method comprises administering an effective amount of the protein according to any one of claims 1-68 or the formulation according to claim 69 to a patient. 73. The method of claim 72, wherein the second antigen binding site of the protein binds c-MET, and the cancer to be treated is selected from the group consisting of renal cancer, thyroid cancer, melanoma, lung cancer, melanoma, liver cancer, pancreatic cancer, colorectal cancer, and head and neck cancer. 74. The method of claim 72, wherein the second antigen binding site of the protein binds KIT, and the cancer to be treated is selected from the group consisting of colorectal cancer, acute myeloid leukemia, gastrointestinal stromal tumor, melanoma, small cell lung cancer, non-small cell lung cancer, renal cancer, liver cancer, and testicular cancer. 75. The method of claim 72, wherein the second antigen binding site of the protein binds F3, and the cancer to be treated is selected from the group consisting of bladder cancer, breast cancer, cervical cancer, glioblastoma, head and neck cancer, lung cancer, pancreatic cancer, prostate cancer, sarcomas, and colorectal cancer. 76. The method of claim 72, wherein the second antigen binding site of the protein binds IGF1R, and the cancer to be treated is selected from the group consisting of breast cancer, cervical cancer, head and neck cancer, lung cancer, melanoma, mesothelioma, ovarian cancer, prostate cancer, sarcoma, thyroid cancer, renal cancer, colorectal cancer, pancreatic cancer, gliobastoma, and liver cancer. 77. The method of claim 72, wherein the second antigen binding site of the protein binds Lewis Y, and the cancer to be treated is selected from the group consisting of ovarian cancer, lung cancer, colorectal cancer, gastric cancer, breast cancer, cervical cancer, head and neck cancer, multiple myeloma, and acute myeloid leukemia. 78. The method of claim 72, wherein the second antigen binding site of the protein binds MUC13, and the cancer to be treated is selected from the group consisting of ovarian cancer, liver cancer, lung cancer, melanoma, liver cancer, gastric cancer, pancreatic cancer, renal cancer, esophageal cancer, breast cancer, colorectal cancer, cervical cancer, and cholangiocarcinoma. 79. The method of claim 72, wherein the second antigen binding site of the protein binds MUC4, and the cancer to be treated is selected from the group consisting of breast cancer, pancreatic cancer, ovarian cancer, lung cancer, acute lymphoblastic leukemia, bladder cancer, cervical cancer, colorectal cancer, head and neck cancer, and prostate cancer. 80. The method of claim 72, wherein the second antigen binding site of the protein binds MCAM, and the cancer to be treated is selected from the group consisting of melanoma, breast cancer, small cell lung cancer, sarcoma, colorectal cancer, pancreatic cancer, and renal cancer. 81. The method of claim 72, wherein the second antigen binding site of the protein binds LRRC32, and the cancer to be treated is selected from the group consisting of renal cancer, pancreatic cancer, sarcoma, ovarian cancer, lung cancer, gliobastoma, head and neck cancer, prostate cancer, liver cancer, breast cancer, and cervical cancer. 82. The method of claim 72, wherein the second antigen binding site of the protein binds sialyl-Tn, and the cancer to be treated is selected from the group consisting of ovarian cancer, pancreatic cancer, bladder cancer, gastric cancer, prostate cancer, colorectal cancer, and breast cancer. 83. The method of claim 72, wherein the second antigen binding site of the protein binds gpA33, and the cancer to be treated is selected from the group consisting of colorectal cancer, gastric cancer, and esophageal cancer. 84. The method of claim 72, wherein the second antigen binding site of the protein binds GD3, and the cancer to be treated is selected from the group consisting of lung cancer, glioma, breast cancer, melanoma, ovarian cancer, pancreatic cancer, and neuroblastoma. 85. The method of claim 72, wherein the second antigen binding site of the protein binds GM2, and the cancer to be treated is selected from the group consisting of gastric cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, neuroblastoma, melanoma, lung cancer, mesothelioma, and liver cancer. 86. The method of claim 72, wherein the second antigen binding site of the protein binds EPHA3, and the cancer to be treated is selected from the group consisting of cervical cancer, head and neck cancer, gastric cancer, multiple myeloma, ovarian cancer, colorectal cancer, melanoma, breast cancer, prostate cancer, pancreatic cancer, lung cancer, and sarcoma. 87. The method of claim 72, wherein the second antigen binding site of the protein binds TNFRSF10A, and the cancer to be treated is selected from the group consisting of breast cancer, colorectal cancer, prostate cancer, pancreatic cancer, bladder cancer, and head and neck cancer. 88. The method of claim 72, wherein the second antigen binding site of the protein binds TNFSF11, and the cancer to be treated is selected from the group consisting of breast cancer, prostate cancer, and a bone metastatic cancer. 89. The method of claim 72, wherein the second antigen binding site of the protein binds CD74, and the cancer to be treated is selected from the group consisting of diffuse large B cell cancer, a B cell malignancy, renal cancer, lung cancer, ovarian cancer, melanoma, sarcoma, head and neck cancer, liver cancer, bladder cancer, glioma, breast cancer, and leukemia. 90. The method of claim 72, wherein the second antigen binding site of the protein binds PMEL, and the cancer to be treated is selected from the group consisting of melanoma and sarcoma. Dr. Shlomo Cohen Co. Law Offices B. S. R Tower 3 5 Kineret Street Bnei Brak 5126237 Tel. 03 - 527 1919
Independent claims12