Multispecific binding proteins that bind dectin-1 and trop-2 and methods of use thereof

HK40137833APending Publication Date: 2026-09-18DREN BIO MANAGEMENT INC
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Patent Information

Application Number
HK62026126469
Authority / Receiving Office
HK · HK
Patent Type
Applications
Current Assignee / Owner
Priority Date
2023-07-31
Filing Date
2026-07-21
Publication Date
2026-09-18
Estimated Expiration
2044-07-29

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Abstract

The present disclosure relates to bispecific binding molecules comprising a first arm comprising a first antigen binding domain that binds to human Dectin-1 and a second arm comprising a second antigen binding domain that binds to human Trop-2, as well as methods, polynucleotides, vectors, host cells, compositions, and uses related thereto
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Description

This disclosure relates to a bispecific binding molecule comprising a first arm and a second arm, the first arm comprising a first antigen-binding domain binding to human Dectin-1, and the second arm comprising a second antigen-binding domain binding to human Trop-2, as well as related methods, polynucleotides, vectors, host cells, compositions, and uses. Abstract

Claims

CLAIMSWhat is claimed is:

1. A bispecific binding molecule, comprising:(a) a first arm comprising a first antigen binding domain that binds to human Dectin- 1; and(b) a second arm comprising a second antigen binding domain that binds to human Trop- 2; wherein the second antigen binding domain comprises a second heavy chain variable (VH) domain and a second light chain variable (VL) domain; and wherein:(a) the second VH domain comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO:37, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:38, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:39, and the second VL domain comprises a CDR-L1 comprising the amino acid sequence of SEQ ID NO:40, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:41, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:42; or(b) the second VH domain comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO:53, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:54, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:55, and the second VL domain comprises a CDR-L1 comprising the amino acid sequence of SEQ ID NO:56, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:57, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 58.

2. A bispecific binding molecule, comprising:(a) a first arm comprising a first antigen binding domain that binds to human Dectin- 1; and(b) a second arm comprising a second antigen binding domain that binds to human Trop- 2;wherein the second antigen binding domain comprises a second heavy chain variable (VH) domain and a second light chain variable (VL) domain; and wherein:(a) the second VH domain comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO:48, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:49, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:50, and the second VL domain comprises a CDR-L1 comprising the amino acid sequence of SEQ ID NO:51, a CDR-L2 comprising the amino acid sequence of GAS, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO: 52; or(b) the second VH domain comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO:64, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:65, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:66, and the second VL domain comprises a CDR-L1 comprising the amino acid sequence of SEQ ID NO:67, a CDR-L2 comprising the amino acid sequence of AAS, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:68.

3. The bispecific binding molecule of claim 1 or claim 2, wherein:(a) the second VH domain comprises the amino acid sequence of SEQ ID NO:74, and the second VL domain comprises the amino acid sequence of SEQ ID NO:75; or(b) the second VH domain comprises the amino acid sequence of SEQ ID NO:79, and the second VL domain comprises the amino acid sequence of SEQ ID NO:80.

4. The bispecific binding molecule of any one of claims 1-3, wherein the second arm comprises an antibody heavy chain polypeptide that comprises the second VH domain and an antibody light chain polypeptide that comprises the second VL domain.

5. The bispecific binding molecule of claim 4, wherein the antibody heavy chain polypeptide of the second arm comprises an Fc region.

6. The bispecific binding molecule of claim 5, wherein the Fc region of the second arm is a human IgG Fc region.

7. The bispecific binding molecule of claim 6, wherein the Fc region of the second arm is a human IgGl Fc region.

8. The bispecific binding molecule of claim 6, wherein the Fc region of the second arm is a human IgG4 Fc region.

9. The bispecific binding molecule of any one of claims 5-8, wherein the first arm comprises an Fc region; wherein the Fc region of the first arm comprises one or more knob-forming mutations, and the Fc region of the second arm comprises one or more cognate hole-forming mutations.

10. The bispecific binding molecule of claim 9, wherein the Fc region of the first arm comprises a T366W substitution, and wherein the Fc region of the second arm comprises T366S, L368A, and Y407V substitutions, according to EU numbering.

11. The bispecific binding molecule of any one of claims 5-8, wherein the first arm comprises an Fc region; wherein the Fc region of the second arm comprises one or more knob-forming mutations, and the Fc region of the first arm comprises one or more cognate hole-forming mutations.

12. The bispecific binding molecule of claim 11, wherein the Fc region of the second arm comprises a T366W substitution, and wherein the Fc region of the first arm comprises T366S, L368A, and Y407V substitutions, according to EU numbering.

13. The bispecific binding molecule of claim 4, wherein:(a) the second arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO:76 or 77 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:78;(b) the second arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO: 81 or 82 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:83;(c) the second arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO: 107 or 108 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:78; or(d) the second arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO: 109 or 110 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:83.

14. A bispecific binding molecule, comprising:(a) a first arm comprising a first antigen binding domain that binds to human Dectin- 1; and(b) a second arm comprising a second antigen binding domain that binds to human Trop- 2; wherein the second arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO:71 or 72 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:73, or wherein the second arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO: 105 or 106 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:73.

15. The bispecific binding molecule of any one of claims 1-14, wherein the first antigen binding domain comprises a first heavy chain variable (VH) domain and a first light chain variable (VL) domain; and wherein the first VH domain comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 1, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:2, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO:3, and the second VL domain comprises a CDR-L1 comprising the amino acid sequence of SEQ ID NO:4, a CDR- L2 comprising the amino acid sequence of SEQ ID NO:5, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:6.

16. The bispecific binding molecule of any one of claims 1-14, wherein the first antigen binding domain comprises a first heavy chain variable (VH) domain and a first light chain variable (VL) domain; and wherein the first VH domain comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 13, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 14, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 15, and the second VL domain comprises a CDR-L1 comprising the amino acid sequence of SEQ ID NO:4, a CDR-L2 comprising the amino acid sequence of SEQ ID NO:5, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:6.

17. The bispecific binding molecule of claim 15 or claim 16, wherein the first VH domain comprises the amino acid sequence of SEQ ID NO:24, and the first VL domain comprises the amino acid sequence of SEQ ID NO:25.

18. The bispecific binding molecule of any one of claims 1-14, wherein the first antigen binding domain comprises a first heavy chain variable (VH) domain and a first light chain variable (VL) domain; and wherein the first VH domain comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 16, a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 17, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 18, and the second VL domain comprises a CDR-L1 comprising the amino acid sequence of SEQ ID NON, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 19, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:20.

19. The bispecific binding molecule of any one of claims 1-14, wherein the first antigen binding domain comprises a first heavy chain variable (VH) domain and a first light chain variable (VL) domain; and wherein the first VH domain comprises a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 13, a CDR-H2 comprising the amino acid sequence of SEQ ID NO:23, and a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 18, and the second VL domain comprises a CDR-L1 comprising the amino acid sequence of SEQ ID NON, a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 19, and a CDR-L3 comprising the amino acid sequence of SEQ ID NO:20.

20. The bispecific binding molecule of claim 18 or claim 19, wherein the first VH domain comprises the amino acid sequence of SEQ ID NO:26, and the first VL domain comprises the amino acid sequence of SEQ ID NO:27.

21. The bispecific binding molecule of claim 18 or claim 19, wherein the first VH domain comprises the amino acid sequence of SEQ ID NO:30, and the first VL domain comprises the amino acid sequence of SEQ ID NON 1.

22. The bispecific binding molecule of any one of claims 1-21, wherein the first arm comprises an antibody heavy chain polypeptide that comprises the first VH domain and an antibody light chain polypeptide that comprises the first VL domain.

23. The bispecific binding molecule of claim 22, wherein the antibody heavy chain polypeptide of the first arm comprises an Fc region.

24. The bispecific binding molecule of any one of claims 1-21, wherein the first arm comprises a single chain variable fragment (scFv) that comprises the first VH domain and the first VL domain.

25. The bispecific binding molecule of claim 24, wherein the scFv comprises a linker between the first VH domain and the first VL domain.

26. The bispecific binding molecule of claim 24 or claim 25, wherein the scFv comprises the amino acid sequence of SEQ ID NO:34.

27. The bispecific binding molecule of any one of claims 24-26, wherein the first arm further comprises an Fc region linked to the scFv.

28. The bispecific binding molecule of claim 23 or claim 27, wherein the Fc region of the first arm is a human IgG Fc region.

29. The bispecific binding molecule of claim 28, wherein the Fc region of the first arm is a human IgGl Fc region.

30. The bispecific binding molecule of claim 28, wherein the Fc region of the first arm is a human IgG4 Fc region.

31. The bispecific binding molecule of any one of claims 28-30, wherein the second arm comprises an Fc region; wherein the Fc region of the first arm comprises one or more knobforming mutations, and the Fc region of the second arm comprises one or more cognate holeforming mutations.

32. The bispecific binding molecule of claim 31, wherein the Fc region of the first arm comprises a T366W substitution, and wherein the Fc region of the second arm comprises T366S, L368A, and Y407V substitutions, according to EU numbering.

33. The bispecific binding molecule of any one of claims 28-30, wherein the second arm comprises Fc region; wherein the Fc region of the second arm comprises one or more knobforming mutations, and the Fc region of the first arm comprises one or more cognate holeforming mutations.

34. The bispecific binding molecule of claim 33, wherein the Fc region of the second arm comprises a T366W substitution, and wherein the Fc region of the first arm comprises T366S, L368A, and Y407V substitutions, according to EU numbering.

35. The bispecific binding molecule of claim 22, wherein the first arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO: 87 or 88 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:89, or wherein the first arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO: 111 or 112 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO: 89.

36. The bispecific binding molecule of claim 22, wherein the first arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO:28 or 90 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:29, or wherein the first arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO:99 or 100 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:29.

37. The bispecific binding molecule of claim 22, wherein the first arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO:32 or 91 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:33, or wherein the first arm comprises an antibody heavy chain polypeptide that comprises the amino acid sequence of SEQ ID NO: 101 or 102 and an antibody light chain polypeptide that comprises the amino acid sequence of SEQ ID NO:33.

38. The bispecific binding molecule of claim 24, wherein the first arm comprises the amino acid sequence of SEQ ID NO:35 or 36, or wherein the first arm comprises the amino acid sequence of SEQ ID NO: 103 or 104.

39. The bispecific binding molecule of any one of claims 1-38, wherein at least one or two of the first and second arms is / are non-fucosylated or comprise(s) reduced fucosylation.

40. The bispecific binding molecule of any one of claims 1-39, wherein human Dectin-1 comprises the amino acid sequence of SEQ ID NO:84 or 85.

41. The bispecific binding molecule of any one of claims 1-40, wherein human Trop-2 comprises the amino acid sequence of SEQ ID NO:86.

42. A bispecific binding molecule, comprising:(a) a first arm comprising a first antigen binding domain that binds to human Dectin- 1; and(b) a second arm comprising a second antigen binding domain that binds to human Trop- 2; wherein the first arm comprises a first antibody heavy chain polypeptide and a first antibody light chain polypeptide, and the second arm comprises a second antibody heavy chain polypeptide and a second antibody light chain polypeptide; and wherein:(a) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:87 or 88, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:89, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:71 or 72, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:73;(b) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:87 or 88, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:89, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:76 or 77, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:78;(c) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:87 or 88, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:89, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 81 or 82, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:83;(d) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:28 or 90, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:29, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:71 or 72, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:73;(e) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:28 or 90, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:29, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:76 or 77, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:78;(f) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:28 or 90, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:29, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 81 or 82, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:83;(g) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:32 or 91, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:33, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:71 or 72, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:73;(h) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:32 or 91, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:33, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:76 or 77, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:78;(i) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:32 or 91, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:33, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 81 or 82, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:83;(j) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 111 or 112, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:89, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 105 or 106, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:73;(k) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 111 or 112, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:89, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 107 or 108, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:78;(l) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 111 or 112, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:89, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 109 or 110, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:83;(m) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:99 or 100, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:29, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 105 or 106, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:73;(n) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:99 or 100, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:29, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 107 or 108, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:78;(o) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:99 or 100, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:29, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 109 or 110, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:83;(p) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 101 or 102, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:33, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 105 or 106, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:73;(q) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 101 or 102, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:33, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 107 or 108, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:78; or(r) the first antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 101 or 102, the first antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:33, the second antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 109 or 110, and the second antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:83.

43. A bispecific binding molecule, comprising:(a) a first arm comprising a first antigen binding domain that binds to human Dectin- 1; and(b) a second arm comprising a second antigen binding domain that binds to human Trop- 2; wherein the first arm comprises a single chain variable fragment (scFv) linked to an Fc region, and the second arm comprises an antibody heavy chain polypeptide and an antibody light chain polypeptide; and wherein:(a) the first arm comprises the amino acid sequence of SEQ ID NO:35 or 36, the antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:71 or 72, and the antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:73;(b) the first arm comprises the amino acid sequence of SEQ ID NO:35 or 36, the antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:76 or 77, and the antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:78;(c) the first arm comprises the amino acid sequence of SEQ ID NO:35 or 36, the antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO:81 or 82, and the antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:83;(d) the first arm comprises the amino acid sequence of SEQ ID NO: 103 or 104, the antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 105 or106, and the antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:73;(e) the first arm comprises the amino acid sequence of SEQ ID NO: 103 or 104, the antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 107 or 108, and the antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:78; or(f) the first arm comprises the amino acid sequence of SEQ ID NO: 103 or 104, the antibody heavy chain polypeptide comprises the amino acid sequence of SEQ ID NO: 109 or 110, and the antibody light chain polypeptide comprises the amino acid sequence of SEQ ID NO:83.

44. A polynucleotide encoding the bispecific binding molecule of any one of claims 1-43.

45. A vector comprising the polynucleotide of claim 44.

46. The vector of claim 45, wherein the vector is an expression vector.

47. An isolated host cell comprising the polynucleotide of claim 44 or the vector of claim 45 or claim 46.

48. The isolated host cell of claim 47, wherein the host cell is a yeast, insect, plant, or prokaryotic cell.

49. The isolated host cell of claim 47, wherein the host cell is a mammalian cell.

50. The isolated host cell of claim 49, wherein the mammalian cell is a Chinese hamster ovary (CHO) cell.

51. The isolated host cell of claim 49 or claim 50, wherein the host cell comprises an alphal,6-fucosyltransferase (Fut8) or alpha-1, 3 -mannosyl-gly coprotein 2-beta-N- acetylglucosaminyltranf erase (MGAT1) knockout.

52. The isolated host cell of claim 49 or claim 50, wherein the host cell overexpresses P 1 ,4- N-acetylglucosaminyltransferase III (GnT-III).

53. The isolated host cell of claim 52, wherein the host cell further overexpresses Golgi p- mannosidase II (Manll).

54. A method of producing a bispecific binding molecule, comprising culturing the host cell of any one of claims 47-53 under conditions suitable for production of the bispecific binding molecule.

55. The method of claim 54, further comprising recovering the bispecific binding molecule from the host cell.

56. The method of claim 54 or claim 55, wherein, prior to production of the bispecific binding molecule, the host cell is treated with kifunensine.

57. A bispecific binding molecule produced by the method of any one of claims 54-56.

58. A pharmaceutical composition comprising the bispecific binding molecule of any one of claims 1-43 and 57 and a pharmaceutically acceptable carrier.

59. A method of treating cancer, comprising administering an effective amount of the bispecific binding molecule of any one of claims 1-43 and 57 or the pharmaceutical composition of claim 58 to an individual in need thereof.

60. The method of claim 59, wherein the cancer is a solid cancer.

61. The method of claim 59 or claim 60, wherein the cancer is breast cancer, prostate cancer, urothelial cancer, cervical cancer, bladder cancer, pancreatic cancer, thyroid cancer, head and neck cancer, esophageal cancer, endometrial cancer, gastric cancer, liver cancer, colorectal cancer, melanoma, lung cancer, ovarian cancer, or uterine cancer.

62. The method of any one of claims 59-61, wherein cells of the cancer express Trop-2.

63. The method of any one of claims 59-62, wherein the individual is a human.