ANTI-SARS-COV-2-SPIKE GLYCOPROTEIN ANTIBODIES AND ANTIGEN-BINDING FRAGMENTS

Abstract

The present disclosure provides antibodies and antigen-binding fragments thereof that bind specifically to a coronavirus spike protein and methods of using such antibodies and fragments for treating or preventing viral infections (e.g., coronavirus infections).

Claims

1. An isolated recombinant antibody or antigen-binding fragment thereof that specifically binds to a coronavirus spike protein (CoV-S), wherein the antibody has one or more of the following characteristics: (a) binds to CoV-S with an EC.sub.50 of less than about 10.sup.8 M; (b) demonstrates an increase in survival in a coronavirus-infected animal after administration to said coronavirus-infected animal, as compared to a comparable coronavirus-infected animal without said administration; and/or (c) comprises three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising an amino acid sequence having at least about 90% sequence identity to an HCVR of Table 4; and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising an amino acid sequence having at least about 90% sequence identity to an LCVR Table 4.

2. The antibody or antigen-binding fragment of claim 1, comprising: (a) a heavy chain variable region comprising the HCDR1, HCDR2, and HCDR3 of an antibody of Table 4; and/or (b) a light chain variable region comprising the LCDR1, LCDR2, and LCDR3 of an antibody of Table 4.

3. The antibody or antigen-binding fragment of claim 1 or 2, comprising: (a) a heavy chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to an HCVR sequence of Table 4; and/or (b) a light chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to an LCVR sequence of Table 4.

4. The antibody or antigen-binding fragment of any one of claims 1-3, wherein said antibody or antigen-binding fragment comprises the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 of a single antibody of Table 4.

5. The antibody or antigen-binding fragment of any one of claims 1-4, comprising an immunoglobulin that comprises the HCVR and the LCVR of a single antibody of Table 4.

6. The antibody or antigen-binding fragment of claim 1, comprising: (a) a heavy chain variable region (HCVR) comprising three complementarity determining regions (CDRs) contained within the amino acid sequence of SEQ ID NO: 212; (b) a HCVR comprising HCDR1, HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 214, 216 and 218, respectively; (c) a HCVR comprising the amino acid sequence of SEQ ID NO: 212; (d) a light chain variable region (LCVR) comprising three CDRs contained within the amino acid sequence of SEQ ID NO: 220; (e) a LCVR comprising LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 222, 126 and 224, respectively; (f) a LCVR comprising the amino acid sequence of SEQ ID NO: 220; (g) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 226; (h) a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 228; (i) a HCVR/LCVR pair comprising the CDRs contained within the amino acid sequences of SEQ ID NOs: 212/222, respectively; (j) a HCVR comprising HCDR1, HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 214, 216 and 218, respectively, and a LCVR comprising LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 222, 126 and 224, respectively; (k) a HCVR comprising the amino acid sequence of SEQ ID NO: 212 and a LCVR comprising the amino acid sequence of SEQ ID NO: 220; or (l) a HC comprising the amino acid sequence of SEQ ID NO: 226 and a LC comprising the amino acid sequence of SEQ ID NO: 228.

7. The antibody or antigen-binding fragment of claim 1, comprising: (a) a heavy chain variable region (HCVR) comprising three complementarity determining regions (CDRs) contained within the amino acid sequence of SEQ ID NO: 362; (b) a HCVR comprising HCDR1, HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 364, 366 and 368, respectively; (c) a HCVR comprising the amino acid sequence of SEQ ID NO: 362; (d) a light chain variable region (LCVR) comprising three CDRs contained within the amino acid sequence of SEQ ID NO: 370; (e) a LCVR comprising LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 372, 106 and 374, respectively; (f) a LCVR comprising the amino acid sequence of SEQ ID NO: 370; (g) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 376; (h) a HC comprising the amino acid sequence of SEQ ID NO: 1077; (i) a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 378; (j) a HCVR/LCVR pair comprising the CDRs contained within the amino acid sequences of SEQ ID NOs: 362/370, respectively; (k) a HCVR comprising HCDR1, HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 364, 366 and 368, respectively, and a LCVR comprising LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 372, 106 and 374 respectively; (l) a HCVR comprising the amino acid sequence of SEQ ID NO: 362 and a LCVR comprising the amino acid sequence of SEQ ID NO: 370; (m) a HC comprising the amino acid sequence of SEQ ID NO: 376 and a LC comprising the amino acid sequence of SEQ ID NO: 378; or (n) a HC comprising the amino acid sequence of SEQ ID NO: 1077 and a LC comprising the amino acid sequence of SEQ ID NO: 378.

8. The antibody or antigen-binding fragment of claim 1, comprising: (a) a heavy chain variable region (HCVR) comprising three complementarity determining regions (CDRs) contained within the amino acid sequence of SEQ ID NO: 493; (b) a HCVR comprising HCDR1, HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 495, 497 and 499, respectively; (c) a HCVR comprising the amino acid sequence of SEQ ID NO: 493; (d) a light chain variable region (LCVR) comprising three CDRs contained within the amino acid sequence of SEQ ID NO: 501; (e) a LCVR comprising LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 503, 505 and 507, respectively; (f) a LCVR comprising the amino acid sequence of SEQ ID NO: 501; (g) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 509; (h) a HC comprising the amino acid sequence of SEQ ID NO: 1075; (i) a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 511; (j) a HCVR/LCVR pair comprising the CDRs contained within the amino acid sequences of SEQ ID NOs: 493/501, respectively; (k) a HCVR comprising HCDR1, HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 495, 497 and 3499 respectively, and a LCVR comprising LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 503, 505 and 507 respectively; (l) a HCVR comprising the amino acid sequence of SEQ ID NO: 493 and a LCVR comprising the amino acid sequence of SEQ ID NO: 501; (m) a HC comprising the amino acid sequence of SEQ ID NO: 509 and a LC comprising the amino acid sequence of SEQ ID NO: 511; or (n) a HC comprising the amino acid sequence of SEQ ID NO: 1075 and a LC comprising the amino acid sequence of SEQ ID NO: 511.

9. The antibody or antigen-binding fragment of claim 1, comprising: (a) a heavy chain variable region (HCVR) comprising three complementarity determining regions (CDRs) contained within the amino acid sequence of SEQ ID NO: 887; (b) a HCVR comprising HCDR1, HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 889, 891 and 893, respectively; (c) a HCVR comprising the amino acid sequence of SEQ ID NO: 887; (d) a light chain variable region (LCVR) comprising three CDRs contained within the amino acid sequence of SEQ ID NO: 895; (e) a LCVR comprising LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 897, 164 and 899, respectively; (f) a LCVR comprising the amino acid sequence of SEQ ID NO: 895; (g) a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 901; (h) a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 903; (i) a HCVR/LCVR pair comprising the CDRs contained within the amino acid sequences of SEQ ID NOs: 887/895, respectively; (j) a HCVR comprising HCDR1, HCDR2 and HCDR3 comprising the amino acid sequences of SEQ ID NOs: 889, 891 and 893, respectively, and a LCVR comprising LCDR1, LCDR2 and LCDR3 comprising the amino acid sequences of SEQ ID NOs: 897, 164 and 899, respectively; (k) a HCVR comprising the amino acid sequence of SEQ ID NO: 887 and a LCVR comprising the amino acid sequence of SEQ ID NO: 895; or (l) a HC comprising the amino acid sequence of SEQ ID NO: 901 and a LC comprising the amino acid sequence of SEQ ID NO: 903.

10. The antibody or antigen-binding fragment of claim 1, comprising: (a) an immunoglobulin heavy chain variable region comprising the HCDR1, HCDR2, and HCDR3 of an antibody of Table 4; and/or (b) an immunoglobulin light chain variable region comprising the LCDR1, LCDR2, and LCDR3 of an antibody of Table 4.

11. An antibody or antigen-binding fragment thereof that competes with the antibody or antigen-binding fragment of any one of the above claims for binding to CoV-S.

12. An antibody or antigen-binding fragment thereof that binds to the same epitope as, or to an overlapping epitope on, CoV-S as the antibody or antigen-binding fragment of any one of the above claims.

13. The antibody or antigen-binding fragment of any one of claims 1-12 which is multispecific.

14. The antibody or antigen-binding fragment of any one of claims 1-13 which comprises one or more of the following properties: (a) inhibits growth of coronavirus; (b) binds to the surface of a coronavirus; (c) limits spread of coronavirus infection of cells in vitro; and (d) protects mice engineered to express the human ACE2 or TMPRSS2 protein from death and/or weight loss caused by coronavirus infection.

15. The antibody or antigen-binding fragment of any one of claims 1-14, wherein said CoV-S is SARS-CoV-2-S.

16. A complex comprising an antibody or antigen-binding fragment of any one of claims 1-15 bound to a CoV-S polypeptide.

17. The complex of claim 16, wherein said CoV-S is SARS-CoV-2-S.

18. A polypeptide comprising: (a) HCDR1, HCDR2, and HCDR3 of an HCVR domain of an antibody or antigen-binding fragment that comprises an HCVR amino acid sequence set forth in Table 4; or (b) LCDR1, LCDR2, and LCDR3 of an LCVR domain of an immunoglobulin chain that comprises an LCVR amino acid sequence set forth in Table 4.

19. A polynucleotide encoding: (a) the polypeptide of claim 18; (b) an antibody or antigen-binding fragment thereof of any one of claims 1-10; (c) a HCVR comprising the CDR sequences contained in a HCVR of Table 4; (d) a LCVR comprising the CDR sequences contained in a LCVR of Table 4; (e) a HCVR comprising HCDR1, HCDR2 and HCDR3 sequences as set forth for an antibody in Table 4; (f) a LCVR comprising LCDR1, LCDR2 and LCDR3 sequences as set forth for an antibody in Table 4; (g) a HCVR and a LCVR comprising CDR sequences contained in an HCVR/LCVR pair of Table 4; (h) a HCVR and a LCVR comprising the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and LCDR3 sequences, respectively, as set forth for an antibody in Table 4; (i) a HCVR comprising the HCVR sequence of an antibody of Table 4; (j) a LCVR comprising the LCVR sequence of an antibody of Table 4; (k) a HCVR and a LCVR comprising the HCVR and LCVR sequences, respectively, of an antibody of Table 4; (l) a heavy chain (HC) comprising the HC sequence of an antibody of Table 4; (m) a light chain (LC) comprising the LC sequence of an antibody of Table 4; or (n) a heavy chain (HC) and a light chain (LC) comprising the HC and LC sequences, respectively, of an antibody of Table 4.

20. A vector comprising the polynucleotide of claim 19.

21. A host cell comprising the antibody or antigen-binding fragment or polypeptide or polynucleotide or vector of any one of claims 1-15, 19 and 20.

22. A pair of polynucleotides, wherein: (a) the first polynucleotide encodes: (i) a HCVR comprising the CDR sequences contained in a HCVR of an antibody of Table 4; (ii) a HCVR comprising the HCDR1, HCDR2 and HCDR3 sequences as set forth for an antibody in Table 4; (iii) a HCVR comprising the HCVR sequence of an antibody of Table 4; or (iv) a heavy chain (HC) comprising the HC sequence of an antibody of Table 4; and (b) the second polynucleotide encodes: (i) a LCVR comprising the CDR sequences contained in a LCVR of an antibody of Table 4; (ii) a LCVR comprising the LCDR1, LCDR2 and LCDR3 sequences as set forth for an antibody in Table 4; (iii) a LCVR comprising the LCVR sequence of an antibody of Table 4; or (iv) a light chain (LC) comprising the LC sequence of an antibody of Table 4.

23. The pair of polynucleotides of claim 22, wherein: (a) the first polynucleotide encodes a HCVR comprising the CDRs contained in a HCVR comprising the amino acid sequence of SEQ ID NO: 212, and the second polynucleotide encodes a LCVR comprising the CDRs contained in a LCVR comprising the amino acid sequence of SEQ ID NO: 220; (b) the first polynucleotide encodes a HCVR comprising the HCDR1, HCDR2 and HCDR2 amino acid sequences of SEQ ID NOs: 214, 216 and 218, respectively, and the second polynucleotide encodes a LCVR comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NOs: 222, 126 and 224, respectively; (c) the first polynucleotide encodes a HCVR comprising the amino acid sequence of SEQ ID NO: 212, and the second polynucleotide encodes a LCVR comprising the amino acid sequence of SEQ ID NO: 220; or (d) the first polynucleotide encodes a HC comprising the amino acid sequence of SEQ ID NO: 226, and the second polynucleotide encodes a LC comprising the amino acid sequence of SEQ ID NO: 228.

24. The pair of polynucleotides of claim 22, wherein: (a) the first polynucleotide encodes a HCVR comprising the CDRs contained in a HCVR comprising the amino acid sequence of SEQ ID NO: 362, and the second polynucleotide encodes a LCVR comprising the CDRs contained in a LCVR comprising the amino acid sequence of SEQ ID NO: 370; (b) the first polynucleotide encodes a HCVR comprising the HCDR1, HCDR2 and HCDR2 amino acid sequences of SEQ ID NOs: 364, 366 and 368, respectively, and the second polynucleotide encodes a LCVR comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NOs: 372, 106 and 374, respectively; (c) the first polynucleotide encodes a HCVR comprising the amino acid sequence of SEQ ID NO: 362, and the second polynucleotide encodes a LCVR comprising the amino acid sequence of SEQ ID NO: 370; (d) the first polynucleotide encodes a HC comprising the amino acid sequence of SEQ ID NO: 376, and the second polynucleotide encodes a LC comprising the amino acid sequence of SEQ ID NO: 378; or (e) the first polynucleotide encodes a HC comprising the amino acid sequence of SEQ ID NO: 1077, and the second polynucleotide encodes a LC comprising the amino acid sequence of SEQ ID NO: 378.

25. The pair of polynucleotides of claim 22, wherein: (a) the first polynucleotide encodes a HCVR comprising the CDRs contained in a HCVR comprising the amino acid sequence of SEQ ID NO: 493, and the second polynucleotide encodes a LCVR comprising the CDRs contained in a LCVR comprising the amino acid sequence of SEQ ID NO: 501; (b) the first polynucleotide encodes a HCVR comprising the HCDR1, HCDR2 and HCDR2 amino acid sequences of SEQ ID NOs: 495, 497 and 499, respectively, and the second polynucleotide encodes a LCVR comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NOs: 503, 505 and 507, respectively; (c) the first polynucleotide encodes a HCVR comprising the amino acid sequence of SEQ ID NO: 493, and the second polynucleotide encodes a LCVR comprising the amino acid sequence of SEQ ID NO: 501; (d) the first polynucleotide encodes a HC comprising the amino acid sequence of SEQ ID NO: 509, and the second polynucleotide encodes a LC comprising the amino acid sequence of SEQ ID NO: 511; or (e) the first polynucleotide encodes a HC comprising the amino acid sequence of SEQ ID NO: 1075, and the second polynucleotide encodes a LC comprising the amino acid sequence of SEQ ID NO: 511.

26. The pair of polynucleotides of claim 22, wherein: (a) the first polynucleotide encodes a HCVR comprising the CDRs contained in a HCVR comprising the amino acid sequence of SEQ ID NO: 887, and the second polynucleotide encodes a LCVR comprising the CDRs contained in a LCVR comprising the amino acid sequence of SEQ ID NO: 895; (b) the first polynucleotide encodes a HCVR comprising the HCDR1, HCDR2 and HCDR2 amino acid sequences of SEQ ID NOs: 889, 891 and 893, respectively, and the second polynucleotide encodes a LCVR comprising the LCDR1, LCDR2 and LCDR3 amino acid sequences of SEQ ID NOs: 897, 164 and 899, respectively; (c) the first polynucleotide encodes a HCVR comprising the amino acid sequence of SEQ ID NO: 887, and the second polynucleotide encodes a LCVR comprising the amino acid sequence of SEQ ID NO: 895; or (d) the first polynucleotide encodes a HC comprising the amino acid sequence of SEQ ID NO: 901, and the second polynucleotide encodes a LC comprising the amino acid sequence of SEQ ID NO: 903.

27. A pair of vectors comprising, respectively, the first and second polynucleotides of any one of claims 22-26.

28. A host cell comprising the pair of vectors of claim 27.

29. A method for making an antibody or antigen-binding fragment of any one of claims 1-15, comprising: (a) introducing into a host cell one or more polynucleotides encoding said antibody or antigen-binding fragment; (b) culturing the host cell under conditions favorable to expression of the one or more polynucleotides; and (c) optionally, isolating the antibody or antigen-binding fragment from the host cell and/or a medium in which the host cell is grown.

30. The method of claim 29, wherein the host cell is a Chinese hamster ovary cell.

31. The method of claim 29 or 30, wherein the one or more polynucleotides is one or more polynucleotides according to claim 19, or the pair of polynucleotides according to any one of claims 22-26.

32. An antibody or antigen-binding fragment which is a product of the method of any one of claims 29-31.

33. A composition or kit comprising the antibody or antigen-binding fragment of any one of claims 1-15 and 32 in association with a further therapeutic agent.

34. A pharmaceutical composition comprising the antigen-binding protein of any one of claims 1-15 and 32 and pharmaceutically acceptable carrier and, optionally, a further therapeutic agent.

35. The composition or kit of claim 33 or 34 in association with a further therapeutic agent which is an anti-viral drug or a vaccine.

36. The composition or kit of any one of claims 33-35 wherein the further therapeutic agent is selected from the group consisting of: an anti-inflammatory agent, an antimalarial agent, an antibody or antigen-binding fragment thereof that specifically binds TMPRSS2, and an antibody or antigen-binding fragment thereof that specifically binds to CoV-S.

37. The composition or kit of claim 36, wherein said antimalarial agent is chloroquine or hydroxychloroquine, and/or said anti-inflammatory agent is an antibody, optionally wherein said antibody is sarilumab, tocilizumab or gimsilumab, and/or said antibody or antigen-binding fragment that specifically binds to CoV-S is casirivimab or imdevimab.

38. The composition or kit of claim 36, wherein the antibody or antigen-binding fragment comprises the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences of a second antibody of Table 4.

39. A vessel or injection device comprising the antigen-binding protein or composition of any one of claims 1-15, 32, and 33-38.

40. A method for treating or preventing infection with a coronavirus, in a subject in need thereof, comprising administering a therapeutically effective amount of antigen-binding protein of any one of claims 1-15 and 32.

41. The method of claim 40, wherein said coronavirus is selected from the group consisting of SARS-CoV-2, SARS-CoV, and MERS-CoV.

42. The method of claim 40 or 41, wherein the subject is administered one or more further therapeutic agents.

43. The method of claim 42, wherein the one or more further therapeutic agents is an anti-viral drug or a vaccine.

44. The method of claim 42, wherein the one or more further therapeutic agents is selected from the group consisting of: an anti-inflammatory agent, an antimalarial agent, an antibody or antigen-binding fragment thereof that specifically binds TMPRSS2, and an antibody or antigen-binding fragment thereof that specifically binds to CoV-S.

45. The method of claim 44, wherein said antimalarial agent is chloroquine or hydroxychloroquine, and/or said anti-inflammatory agent is an antibody, optionally wherein said antibody is sarilumab, tocilizumab, or gimsilumab, and/or said antibody or antigen-binding fragment that specifically binds to CoV-S is casirivimab or imdevimab.

46. The method of claim 44, wherein the antibody or antigen-binding fragment comprises the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences of a second antibody of Table 4.

47. A method for administering an antibody or antigen-binding fragment of any one of claims 1-15 and 32 into the body of a subject comprising injecting the antibody or antigen-binding fragment into the body of the subject.

48. The method of claim 47, wherein the antibody or antigen-binding fragment is injected into the body of the subject subcutaneously, intravenously or intramuscularly.

49. The antibody or antigen-binding fragment, composition, kit, complex, polypeptide, polynucleotide, vector, cell, or method of any of the above claims, wherein said antibody or antigen-binding binding fragment comprises a VH3-66 or Vk1-33 variable domain sequence.

50. An isolated antibody or antigen-binding fragment thereof that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, wherein said isolated antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 887, and three light chain complementarity determining regions (CDRs) (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 895.

51. The isolated antibody or antigen-binding fragment thereof of claim 50, wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 889, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 891, HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 893, LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 897, LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 164, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 899.

52. The isolated antibody or antigen-binding fragment thereof of claim 51, comprising an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 887.

53. The isolated antibody or antigen-binding fragment thereof of claim 51, comprising an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 895.

54. The isolated antibody or antigen-binding fragment thereof of claim 51, comprising an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 887 and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 895.

55. An isolated antibody that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, wherein said isolated antibody comprises an immunoglobulin constant region, three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 887, and three light chain complementarity determining regions (CDRs) (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 895.

56. The isolated antibody of claim 55, wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 889, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 891, HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 893, LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 897, LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 164, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 899.

57. The isolated antibody of claim 55, comprising an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 887 and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 895.

58. The isolated antibody of claim 55, wherein said isolated antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 901 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 903.

59. The isolated antibody of claim 55, wherein said immunoglobulin constant region is an IgG1 constant region.

60. The isolated antibody of claim 55 which is a recombinant antibody.

61. The isolated antibody of claim 55 which is multispecific.

62. A pharmaceutical composition comprising the isolated antibody of claim 55 and a pharmaceutically acceptable carrier or diluent.

63. The pharmaceutical composition of claim 62, further comprising a second therapeutic agent.

64. The pharmaceutical composition of claim 63, wherein the second therapeutic agent is selected from the group consisting of: a second antibody, or an antigen-binding fragment thereof, that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, an anti-inflammatory agent, an antimalarial agent, and an antibody or antigen-binding fragment thereof that binds TMPRSS2.

65. The pharmaceutical composition of claim 63, wherein the second therapeutic agent is a second antibody, or an antigen-binding fragment thereof, that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008.

66. The pharmaceutical composition of claim 65, wherein the second antibody or antigen-binding fragment thereof comprises three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within an HCVR comprising the amino acid sequence set forth in SEQ ID NO: 212, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within an LCVR comprising the amino acid sequence set forth in SEQ ID NO: 220.

67. The pharmaceutical composition of claim 66, wherein the second antibody or antigen-binding fragment thereof comprises: HCDR1, comprising the amino acid sequence set forth in SEQ ID NO: 214; HCDR2, comprising the amino acid sequence set forth in SEQ ID NO: 216; HCDR3, comprising the amino acid sequence set forth in SEQ ID NO: 218; LCDR1, comprising the amino acid sequence set forth in SEQ ID NO: 222; LCDR2, comprising the amino acid sequence set forth in SEQ ID NO: 126; and LCDR3, comprising the amino acid sequence set forth in SEQ ID NO: 224.

68. The pharmaceutical composition of claim 67, wherein the second antibody or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence set forth in SEQ ID NO: 212 and an LCVR comprising the amino acid sequence set forth in SEQ ID NO: 220.

69. The pharmaceutical composition of claim 68, wherein the second antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 226 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 228.

70. An isolated antibody or antigen-binding fragment thereof that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, wherein said isolated antibody or antigen-binding fragment comprises three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 212, and three light chain complementarity determining regions (CDRs) (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 220.

71. The isolated antibody or antigen-binding fragment thereof of claim 70, wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 214, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 216, HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 218, LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 222, LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 126, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 224.

72. The isolated antibody or antigen-binding fragment thereof of claim 70, comprising an HCVR that comprises an amino acid sequence set forth in SEQ ID NO: 212.

73. The isolated antibody or antigen-binding fragment thereof of claim 70, comprising an LCVR that comprises an amino acid sequence set forth in SEQ ID NO: 220.

74. The isolated antibody or antigen-binding fragment thereof of claim 70, comprising an HCVR that comprises an amino acid sequence set forth in SEQ ID NO: 212 and an LCVR that comprises an amino acid sequence set forth in SEQ ID NO: 220.

75. An isolated antibody that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, wherein said isolated antibody comprises an immunoglobulin constant region, three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 212, and three light chain complementarity determining regions (CDRs) (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 220.

76. The isolated antibody of claim 75, wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 214, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 216, HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 218, LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 222, LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 126, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 224.

77. The isolated antibody of claim 75, comprising an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 212 and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 220.

78. The isolated antibody of claim 75, wherein said isolated antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 226 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 228.

79. The isolated antibody of claim 75, wherein said immunoglobulin constant region is an IgG1 constant region.

80. The isolated antibody of claim 75, which is a recombinant antibody.

81. The isolated antibody of claim 75, which is multispecific.

82. A pharmaceutical composition comprising the isolated antibody of claim 75 and a pharmaceutically acceptable carrier or diluent.

83. The pharmaceutical composition of claim 82, further comprising a second therapeutic agent.

84. The pharmaceutical composition of claim 83, wherein the second therapeutic agent is selected from the group consisting of: a second antibody, or an antigen-binding fragment thereof, that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, an anti-inflammatory agent, an antimalarial agent, and an antibody or antigen-binding fragment thereof that binds TMPRSS2.

85. The pharmaceutical composition of claim 83, wherein the second therapeutic agent is a second antibody, or an antigen-binding fragment thereof, that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008.

86. The pharmaceutical composition of claim 85, wherein the second antibody or antigen-binding fragment thereof comprises three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within an HCVR comprising the amino acid sequence set forth in SEQ ID NO: 887, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within an LCVR comprising the amino acid sequence set forth in SEQ ID NO: 895.

87. The pharmaceutical composition of claim 86, wherein the second antibody or antigen-binding fragment thereof comprises: HCDR1, comprising the amino acid sequence set forth in SEQ ID NO: 889; HCDR2, comprising the amino acid sequence set forth in SEQ ID NO: 891; HCDR3, comprising the amino acid sequence set forth in SEQ ID NO: 893; LCDR1, comprising the amino acid sequence set forth in SEQ ID NO: 897; LCDR2, comprising the amino acid sequence set forth in SEQ ID NO: 164; and LCDR3, comprising the amino acid sequence set forth in SEQ ID NO: 899.

88. The pharmaceutical composition of claim 87, wherein the second antibody or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence set forth in SEQ ID NO: 887 and an LCVR comprising the amino acid sequence set forth in SEQ ID NO: 895.

89. The pharmaceutical composition of claim 88, wherein the second antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 901 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 903.

90. An isolated antibody that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, wherein said isolated antibody comprises an immunoglobulin constant region, three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 270, and three light chain complementarity determining regions (CDRs) (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 278.

91. The isolated antibody of claim 90, wherein HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 272, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 274, HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 276, LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 280, LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 106, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 282.

92. The isolated antibody of claim 90, comprising an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 270 and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 278.

93. The isolated antibody of claim 90, wherein said isolated antibody comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 284 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 286.

94. The isolated antibody of claim 90, wherein said immunoglobulin constant region is an IgG1 constant region.

95. The isolated antibody of claim 90 which is a recombinant antibody.

96. The isolated antibody of claim 90 which is multispecific.

97. A pharmaceutical composition comprising the isolated antibody of claim 90 and a pharmaceutically acceptable carrier or diluent.

98. The pharmaceutical composition of claim 97, further comprising a second therapeutic agent.

99. The pharmaceutical composition of claim 98, wherein the second therapeutic agent is selected from the group consisting of: a second antibody, or an antigen-binding fragment thereof, that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, an anti-inflammatory agent, an antimalarial agent, and an antibody or antigen-binding fragment thereof that binds TMPRSS2.

100. The pharmaceutical composition of claim 98, wherein the second therapeutic agent is a second antibody, or an antigen-binding fragment thereof, that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008.

101. The pharmaceutical composition of claim 100, wherein the second antibody or antigen-binding fragment thereof comprises three heavy chain CDRs (HCDR1, HCDR2 and HCDR3) contained within an HCVR comprising the amino acid sequence set forth in SEQ ID NO: 212, and three light chain CDRs (LCDR1, LCDR2 and LCDR3) contained within an LCVR comprising the amino acid sequence set forth in SEQ ID NO: 220.

102. The pharmaceutical composition of claim 101, wherein the second antibody or antigen-binding fragment thereof comprises: HCDR1, comprising the amino acid sequence set forth in SEQ ID NO: 214; HCDR2, comprising the amino acid sequence set forth in SEQ ID NO: 216; HCDR3, comprising the amino acid sequence set forth in SEQ ID NO: 218; LCDR1, comprising the amino acid sequence set forth in SEQ ID NO: 222; LCDR2, comprising the amino acid sequence set forth in SEQ ID NO: 126; and LCDR3, comprising the amino acid sequence set forth in SEQ ID NO: 224.

103. The pharmaceutical composition of claim 102, wherein the second antibody or antigen-binding fragment thereof comprises an HCVR comprising the amino acid sequence set forth in SEQ ID NO: 212 and an LCVR comprising the amino acid sequence set forth in SEQ ID NO: 220.

104. The pharmaceutical composition of claim 103, wherein the second antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 226 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 228.

105. A pharmaceutical composition comprising: a) a first isolated antibody that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, wherein said first isolated antibody comprises an immunoglobulin constant region, three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 887, and three light chain complementarity determining regions (CDRs) (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 895; and b) a second isolated antibody that binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, wherein said second isolated antibody comprises an immunoglobulin constant region, three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 1030, and three light chain complementarity determining regions (CDRs) (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 1038.

106. The pharmaceutical composition of claim 105, wherein the first isolated antibody comprises: HCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 889, HCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 891, HCDR3 comprising the amino acid sequence set forth in SEQ ID NO: 893, LCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 897, LCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 164, and LCDR3 comprising the amino acid sequence set forth in SEQ ID NO: 899.

107. The pharmaceutical composition of claim 105, wherein the first isolated antibody comprises: an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 887 and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 895.

108. The pharmaceutical composition of claim 105, wherein the first isolated antibody comprises: a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 901 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 903.

109. The pharmaceutical composition of claim 105, wherein the first isolated antibody comprises: an immunoglobulin constant region that is an IgG1 constant region.

110. The pharmaceutical composition of claim 105, wherein the first isolated antibody is a recombinant antibody.

111. The pharmaceutical composition of claim 105, wherein the first isolated antibody is multispecific.

112. The pharmaceutical composition of claim 105, further comprising a pharmaceutically acceptable carrier or diluent.

113. The pharmaceutical composition of claim 105, wherein the second isolated antibody comprises: HCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1032, HCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 1034, HCDR3 comprising the amino acid sequence set forth in SEQ ID NO: 1036, LCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1040, LCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 1042, and LCDR3 comprising the amino acid sequence set forth in SEQ ID NO: 1044.

114. The pharmaceutical composition of claim 105, wherein the second isolated antibody comprises: an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1030 and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1038.

115. The pharmaceutical composition of claim 105, wherein the second isolated antibody comprises: a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 1048 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 1048.

116. The pharmaceutical composition of any one of claims 105-115, wherein said pharmaceutical composition further comprises a third isolated antibody.

117. The pharmaceutical composition of claim 116, wherein the third isolated antibody binds a SARS-CoV-2 spike protein comprising the amino acid sequence set forth in SEQ ID NO: 1008, wherein said third isolated antibody comprises an immunoglobulin constant region, three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2 and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 1010, and three light chain complementarity determining regions (CDRs) (LCDR1, LCDR2 and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 1018.

118. The pharmaceutical composition of claim 116, wherein the third isolated antibody comprises: HCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1012, HCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 1014, HCDR3 comprising the amino acid sequence set forth in SEQ ID NO: 1016, LCDR1 comprising the amino acid sequence set forth in SEQ ID NO: 1020, LCDR2 comprising the amino acid sequence set forth in SEQ ID NO: 1022, and LCDR3 comprising the amino acid sequence set forth in SEQ ID NO: 1024.

119. The pharmaceutical composition of claim 116, wherein the third isolated antibody comprises: an HCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1010 and an LCVR that comprises the amino acid sequence set forth in SEQ ID NO: 1018.

120. The pharmaceutical composition of claim 116, wherein the third isolated antibody comprises: a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 1026 and a light chain comprising the amino acid sequence set forth in SEQ ID NO: 1028.

121. A method for treating or preventing infection with an Omicron variant SARS-CoV-2, in a subject in need thereof, comprising administering a therapeutically effective amount of an antibody or antigen binding fragment thereof comprising three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising an amino acid sequence having at least about 90% sequence identity to an HCVR of Table 4; and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising an amino acid sequence having at least about 90% sequence identity to an LCVR Table 4.

122. A method for preventing one or more COVID-19 symptoms resulting from infection with an Omicron variant SARS-CoV-2, in a subject in need thereof, comprising administering a therapeutically effective amount of an antibody or antigen binding fragment thereof comprising three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising an amino acid sequence having at least about 90% sequence identity to an HCVR of Table 4; and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising an amino acid sequence having at least about 90% sequence identity to an LCVR Table 4.

123. The method of claim 122, wherein said preventing comprises pre-exposure prophylaxis.

124. The method of claim 122, wherein said preventing comprises post-exposure prophylaxis.

125. The method of any one of claims 121-124, wherein said antibody or antigen-binding fragment comprises: (a) an immunoglobulin heavy chain variable region comprising the HCDR1, HCDR2, and HCDR3 of an antibody of Table 4; and/or (b) an immunoglobulin light chain variable region comprising the LCDR1, LCDR2, and LCDR3 of an antibody of Table 4.

126. The method of any one of claims 121-124, wherein said antibody or antigen-binding fragment comprises the HCDR1, HCDR2, and HCDR3 of an antibody of Table 4 and the LCDR1, LCDR2, and LCDR3 of said antibody of Table 4.

127. The method of any one of claims 121-126, wherein the subject is administered one or more further therapeutic agents.

128. The method of claim 127, wherein the one or more further therapeutic agents is an anti-viral drug or a vaccine.

129. The method of claim 127, wherein the one or more further therapeutic agents is selected from the group consisting of: an anti-inflammatory agent, an antimalarial agent, an antibody or antigen-binding fragment thereof that specifically binds TMPRSS2, and an antibody or antigen-binding fragment thereof that specifically binds to CoV-S.

130. The method of claim 129, wherein said antimalarial agent is chloroquine or hydroxychloroquine, and/or said anti-inflammatory agent is an antibody, optionally wherein said antibody is sarilumab, tocilizumab, or gimsilumab, and/or said antibody or antigen-binding fragment that specifically binds to CoV-S is casirivimab or imdevimab.

131. The method of claim 129, wherein the antibody or antigen-binding fragment comprises the HCDR1, HCDR2, HCDR3, LCDR1, LCDR2, and LCDR3 sequences of a second antibody of Table 4.

132. The method of any one of claims 121-131, wherein said administering comprises injecting the antibody or antigen-binding fragment into the body of the subject.

133. The method of claim 132, wherein the antibody or antigen-binding fragment is injected into the body of the subject subcutaneously, intravenously or intramuscularly.

134. A method for treating or preventing one or more COVID-19 symptoms resulting from infection with SARS-CoV-2, in a subject in need thereof, comprising administering a therapeutically effective amount of an antibody or antigen binding fragment thereof comprising three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 362; and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 370.

135. The method of claim 134, wherein said HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 364, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 366, HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 368, LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 372, LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 106, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 374.

136. The method of any one of claims 134 and 135, wherein said HCVR comprises the amino acid sequence set forth in SEQ ID NO: 362.

137. The method of any one of claims 134-136, wherein said LCVR comprises the amino acid sequence set forth in SEQ ID NO: 370.

138. The method of any one of claims 134-137, wherein said antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 376.

139. The method of any one of claims 134-138, wherein said antibody or antigen-binding fragment thereof comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 378.

140. The method of any one of claims 134-139, wherein said administering comprises administering said antibody or antigen-binding fragment via injection.

141. The method of claim 140, wherein said injection is intravenous.

142. The method of claim 140, wherein said injection is subcutaneous.

143. The method of any one of claims 134-142, wherein said administering comprises administering 300 mg of said antibody or antigen-binding fragment thereof.

144. The method of any one of claims 134-142, wherein said administering comprises administering 600 mg of said antibody or antigen-binding fragment thereof.

145. The method of any one of claims 134-142, wherein said administering comprises administering 1200 mg of said antibody or antigen-binding fragment thereof.

146. The method of any one of claims 134-142, wherein said administering comprises administering two doses of said antibody or antigen-binding fragment thereof.

147. The method of claim 146, wherein each of said two doses comprises 300 mg of said antibody or antigen-binding fragment thereof.

148. The method of claim 146 or 147, wherein said two doses are administered 4 weeks apart, 5 weeks apart, 6 weeks apart, 7 weeks apart, 8 weeks apart, 9 weeks apart, 10 weeks apart, 11 weeks apart, 12 weeks apart, 13 weeks apart, 14 weeks apart, 15 weeks apart, or 16 weeks apart.

149. The method of claim 146 or 147, wherein said two doses are administered 8-16 weeks apart.

150. The method of claim 146 or 147, wherein said two doses are administered 12 weeks apart.

151. The method of claim 146 or 147, wherein said two doses are administered 8 weeks apart.

152. The method of any one of claims 134-151, wherein said administering reduces the viral load of SARS-CoV-2 in said subject.

153. The method of any one of claims 134-151, wherein said administering occurs prior to SARS-CoV-2 infection.

154. A method for treating or preventing one or more COVID-19 symptoms resulting from infection with SARS-CoV-2, in a subject in need thereof, comprising administering a therapeutically effective amount of an antibody or antigen binding fragment thereof comprising three heavy chain complementarity determining regions (CDRs) (HCDR1, HCDR2, and HCDR3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 362; and three light chain CDRs (LCDR1, LCDR2, and LCDR3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 370, wherein said administering comprises administering to said subject two doses of said antibody or antigen-binding fragment thereof, each does comprising 300 mg of said antibody or antigen-binding fragment thereof.

155. The method of claim 154, wherein said administering is subcutaneous.

156. The method of claim 154 or 155, wherein said HCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 364, HCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 366, HCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 368, LCDR1 comprises the amino acid sequence set forth in SEQ ID NO: 372, LCDR2 comprises the amino acid sequence set forth in SEQ ID NO: 106, and LCDR3 comprises the amino acid sequence set forth in SEQ ID NO: 374.

157. The method of any one of claims 154-156, wherein said HCVR comprises the amino acid sequence set forth in SEQ ID NO: 362.

158. The method of any one of claims 154-157, wherein said LCVR comprises the amino acid sequence set forth in SEQ ID NO: 370.

159. The method of any one of claims 154-158, wherein said antibody or antigen-binding fragment thereof comprises a heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 376.

160. The method of any one of claims 154-158, wherein said antibody or antigen-binding fragment thereof comprises a light chain comprising the amino acid sequence set forth in SEQ ID NO: 378.

161. The method of any one of claims 154-160, wherein said administering comprises administering 600 mg of said antibody or antigen-binding fragment thereof.

162. The method of any one of claims 154-160, wherein said administering comprises administering 300 mg of said antibody or antigen-binding fragment thereof.

163. A polynucleotide encoding: (a) the isolated antibody or antigen-binding fragment thereof of any one of claims 41-52, 61-72 or 81-87; or (b) a HCVR and/or a LCVR; or a heavy chain and/or a light chain of an antibody or antigen-binding fragment thereof of any one of claims 50-61, 70-81 or 90-96.

164. A vector comprising the polynucleotide of claim 163.

165. A host cell comprising the antibody or antigen-binding fragment thereof or polynucleotide or vector of any one of claims 50-61, 70-81, 90-96, 163 or 164.

166. The antibody or antigen-binding fragment thereof of any one of claims 1-15, 32, 50-61, 70-81 or 90-96 that neutralizes an omicron variant of SARS-CoV-2.

167. The antibody or antigen-binding fragment thereof of claim 166, wherein the omicron variant is selected from BA.1, BA.1.1, BA.2, BA.2.12.1, BA.3, or BA.4/BA.5.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0080] FIG. 1 shows cross-competition between 41 anti-SARS-CoV-2-S monoclonal antibodies upon binding to immobilized SARS-Cov-2 RBD-MMH (SEQ ID NO: 1069). Red: Pre-bound mAb-1 reduced binding of mAb-2 to SARS-CoV-2 RBD-MMH by greater than 50%, and binding to SARS-CoV-2 RBD-MMH was also reduced by greater than 50% when the binding order of mAb-1 and mAb-2 was reversed. Yellow: Pre-bound mAb-1 reduced binding of mAb-2 to SARS-CoV-2 RBD-MMH by greater than 50%, but binding to SARS-CoV-2 RBD-MMH was reduced by less than 50% when the binding order of mAb-1 and mAb-2 was reversed.

[0081] FIG. 2 shows cross-competition between 15 anti-SARS-CoV-2-S monoclonal antibodies upon binding to immobilized SARS-Cov-2 RBD-MMH. Red: Pre-bound mAb-1 reduced binding of mAb-2 to SARS-CoV-2 RBD-MMH by greater than 50%, and binding to SARS-CoV-2 RBD-MMH was also reduced by greater than 50% when the binding order of mAb-1 and mAb-2 was reversed. Yellow: Pre-bound mAb-1 reduced binding of mAb-2 to SARS-CoV-2 RBD-MMH by greater than 50%, but binding to SARS-CoV-2 RBD-MMH was reduced by less than 50% when the binding order of mAb-1 and mAb-2 was reversed.

[0082] FIG. 3 depicts the cryo-EM structure of mAb14256, mAb10987, and the receptor binding domain (RBD) of the SARS-CoV-2 spike glycoprotein, in complex, at 3.9 resolution.

[0083] FIG. 4 shows that mAb14256 binds at the top of the RBD, thereby blocking ACE2 binding. mAb14256 competes with mAb10933 (middle structure) and mAb10985 (not depicted).

[0084] FIG. 5 depicts the cryo-EM structure of mAb15160, mAb14315, and the receptor binding domain (RBD) of the SARS-CoV-2 spike glycoprotein, in complex, at 3.18 resolution.

[0085] FIG. 6 depicts the cryo-EM structure of antigen-binding fragments of mAb1428 and mAb15160 with the receptor binding domain (RBD) from the Wuhan-Hu-1 strain and BA.1 lineage, in complex, at 3.3 and 3.4 resolution, respectively.

DETAILED DESCRIPTION OF THE INVENTION

[0086] Before the present methods are described, it is to be understood that this invention is not limited to particular methods, and experimental conditions described, as such methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0087] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference in their entirety.

[0088] The term coronavirus or CoV refers to any virus of the coronavirus family, including but not limited to SARS-CoV-2, MERS-CoV, and SARS-CoV. SARS-CoV-2 has also been known as 2019-nCoV and Wuhan coronavirus. It binds via the viral spike protein to human host cell receptor angiotensin-converting enzyme 2 (ACE2). The spike protein also binds to and is cleaved by TMPRSS2, which activates the spike protein for membrane fusion of the virus.

[0089] The term CoV-S, also called S or S protein refers to the spike protein of a coronavirus, and can refer to specific S proteins such as SARS-CoV-2-S, MERS-CoV S, and SARS-CoV S. The SARS-CoV-2-Spike protein is a 1273 amino acid type I membrane glycoprotein which assembles into trimers that constitute the spikes or peplomers on the surface of the enveloped coronavirus particle. The protein has two essential functions, host receptor binding and membrane fusion, which are attributed to the N-terminal (S1) and C-terminal (S2) halves of the S protein. CoV-S binds to its cognate receptor via a receptor binding domain (RBD) present in the S1 subunit. The amino acid sequence of full-length SARS-CoV-2 spike protein is exemplified by the amino acid sequence provided in SEQ ID NO: 1008. The term CoV-S includes protein variants of CoV spike protein isolated from different CoV isolates as well as recombinant CoV spike protein or a fragment thereof. The term also encompasses CoV spike protein or a fragment thereof coupled to, for example, a histidine tag, mouse or human Fc, or a signal sequence such as ROR1.

[0090] The term coronavirus infection or CoV infection, as used herein, refers to infection with a coronavirus such as SARS-CoV-2, MERS-CoV, or SARS-CoV. The term includes coronavirus respiratory tract infections, often in the lower respiratory tract. Symptoms can include high fever, dry cough, shortness of breath, pneumonia, gastro-intestinal symptoms such as diarrhea, organ failure (kidney failure and renal dysfunction), septic shock, and death in severe cases.

Viruses

[0091] The present invention includes methods for treating or preventing a viral infection in a subject. The term virus includes any virus whose infection in the body of a subject is treatable or preventable by administration of an anti-CoV-S antibody or antigen-binding fragment thereof (e.g., wherein infectivity of the virus is at least partially dependent on CoV-S). In an embodiment of the invention, a virus is any virus that expresses spike protein (e.g., CoV-S). The term virus also includes a CoV-S-dependent respiratory virus which is a virus that infects the respiratory tissue of a subject (e.g., upper and/or lower respiratory tract, trachea, bronchi, lungs) and is treatable or preventable by administration of an anti-CoV-S antibody or antigen-binding fragment thereof. For example, in an embodiment of the invention, virus includes coronavirus, SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2), SARS-CoV (severe acute respiratory syndrome coronavirus), and MERS-CoV (Middle East respiratory syndrome (MERS) coronavirus). Coronaviruses can include the genera of alphacoronaviruses, betacoronaviruses, gammacoronaviruses, and deltacoronaviruses. In some embodiments, the antibodies or antigen-binding fragments provided herein can bind to and/or neutralize an alphacoronavirus, a betacoronavirus, a gammacoronavirus, and/or a deltacoronavirus. In certain embodiments, this binding and/or neutralization can be specific for a particular genus of coronavirus or for a particular subgroup of a genus. Viral infection refers to the invasion and multiplication of a virus in the body of a subject.

[0092] Coronavirus virions are spherical with diameters of approximately 125 nm. The most prominent feature of coronaviruses is the club-shape spike projections emanating from the surface of the virion. These spikes are a defining feature of the virion and give them the appearance of a solar corona, prompting the name, coronaviruses. Within the envelope of the virion is the nucleocapsid. Coronaviruses have helically symmetrical nucleocapsids, which is uncommon among positive-sense RNA viruses, but far more common for negative-sense RNA viruses. SARS-CoV-2, MERS-CoV, and SARS-CoV belong to the coronavirus family. The initial attachment of the virion to the host cell is initiated by interactions between the S protein and its receptor. The sites of receptor binding domains (RBD) within the S1 region of a coronavirus S protein vary depending on the virus, with some having the RBD at the C-terminus of S1. The S-protein/receptor interaction is the primary determinant for a coronavirus to infect a host species and also governs the tissue tropism of the virus. Many coronaviruses utilize peptidases as their cellular receptor. Following receptor binding, the virus must next gain access to the host cell cytosol. This is generally accomplished by acid-dependent proteolytic cleavage of S protein by a cathepsin, TMPRRS2 or another protease, followed by fusion of the viral and cellular membranes.

[0093] The coronaviruses described herein also include variant coronaviruses, which in some embodiments are classified as variants of interest or variants of concern by the World Health Organization (WHO). Variant coronaviruses can have mutations in their spike glycoprotein, which may change the virus's properties such as severity of COVID-19 or transmissibility. WHO defines a variant of concern as one that is associated with one or more of the following changes from wild-type, to such a degree that it is a matter of global public health significance: 1) increased transmissibility or detrimental change in COVID-19 epidemiology; 2) increased virulence or changed clinical disease presentation; or 3) decreased effectiveness of public health measures and social measures, including available therapeutics, vaccines, and diagnostics. Variants of interest are defined by WHO as SARS-CoV-2 viruses 1) with genetic changes that are predicted or have been demonstrated to affect virus characteristics including immune escape, therapeutic escape, diagnostic escpe, transmissibility, and disease severity; and 2) that have been identified as causing significant community transmission or multiple COVID-19 clusters, in multiple countries and with increasing relative prevalence alongside an increasing number of cases over time, or with other epidemiological impacts that suggest an emerging risk to global public health. As of Dec. 13, 2021, there are five variants of concern as classified by WHO (alpha, beta, gamma, delta, and omicron), and two variants of interest as classified by WHO (lambda and mu). Each variant can be defined by the mutations in its spike glycoprotein as compared to the wild-type spike glycoprotein. For example, the Omicron variant (also classified as B.1.1.529) comprises the following mutations in its spike glycoprotein: A67V, 69-70, T95I, G142D/143-145, 211/L212I, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, and L981F (full length omicron spike glycoprotein: SEQ ID NO: 1072). Moreover, these variants may have further lineages that encompass a group of related viruses. For example, as of Jul. 8, 2022, the Omicron variant has BA.1, BA.1.1, BA.2, BA.2.12.1, BA.3, and BA.4/BA.5 lineages. In some embodiments, the antibodies and antigen-binding fragments thereof described herein can bind to any of these variants and/or lineages. In further embodiments, the antibodies and antigen-binding fragments thereof can neutralize these variants and/or lineages.

Anti-CoV-S Antibodies and Antigen-Binding Fragments

[0094] The present invention provides antigen-binding proteins, such as antibodies and antigen-binding fragments thereof, that specifically bind to CoV spike protein or an antigenic fragment thereof.

[0095] The term antibody, as used herein, refers to immunoglobulin molecules comprising four polypeptide chains, two heavy chains (HCs) and two light chains (LCs) inter-connected by disulfide bonds (i.e., full antibody molecules), as well as multimers thereof (e.g. IgM). Exemplary antibodies include, for example, those listed in Table 4. Each heavy chain comprises a heavy chain variable region (HCVR or V.sub.H) and a heavy chain constant region (comprised of domains C.sub.H1, C.sub.H2 and C.sub.H3). Each light chain is comprised of a light chain variable region (LCVR or V.sub.L) and a light chain constant region (C.sub.L). The V.sub.H and V.sub.L regions can be further subdivided into regions of hypervariability, termed complementarity determining regions (CDR), interspersed with regions that are more conserved, termed framework regions (FR). Each V.sub.H and V.sub.L comprises three CDRs and four FRs, arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. Heavy chain CDRs can also be referred to as HCDRs or CDR-Hs, and numbered as described above (e.g., HCDR1, HCDR2, and HCDR3 or HCDR1, HCDR2, and HCDR3). Likewise, light chain CDRs can be referred to as LCDRs or CDR-Ls, and numbered LCDR1, LCDR2, and LCDR3, or LCDR1, LCDR2, and LCDR3. In certain embodiments of the invention, the FRs of the antibody (or antigen binding fragment thereof) are identical to the human germline sequences, or are naturally or artificially modified. Exemplary human germline sequences include, but are not limited to, VH3-66 and Vk1-33. Thus, the present disclosure provides anti-CoV-S antibodies or antigen-binding fragments thereof (e.g., anti-SARS-CoV-2-S antibodies or antigen-binding fragments thereof) comprising HCDR and LCDR sequences of Table 4 within a VH3-66 or Vk1-33 variable heavy chain or light chain region. The present disclosure further provides anti-CoV-S antibodies or antigen-binding fragments thereof (e.g., anti-SARS-CoV-2-S antibodies or antigen-binding fragments thereof) comprising HCDR and LCDR sequences of Table 4 within a combination of a light chain selected from IgKV4-1, IgKV1-5, IgKV1-9, IgKV1-12, IgKV3-15, IgKV1-16, IgKV1-17, IgKV3-20, IgLV3-21, IgKV2-24, IgKV1-33, IgKV1-39, IgLV1-40, IgLV1-44, IgLV1-51, IgLV3-1, IgKV1-6, IgLV2-8, IgKV3-11, IgLV2-11, IgLV2-14, IgLV2-23, or IgLV6-57, and a heavy chain selected from IgHV1-69, IgHV3-64, IgHV4-59, IgHV3-53, IgHV3-48, IgHV4-34, IgHV3-33, IgHV3-30, IgHV3-23, IgHV3-20, IgHV1-18, IgHV3-15, IgHV3-11, IgHV3-9, IgHV1-8, IgHV3-7, IgHV2-5, IgHV1-2, IgHV2-70, IgHV3-66, IgHV5-51, IgHV1-46, IgHV4-39, IgHV4-31, IgHV3-30-3, IgHV2-26, or IgHV7-4-1. The present disclosure further provides anti-CoV-S antibodies or antigen-binding fragments thereof (e.g., anti-SARS-CoV-2-S antibodies or antigen-binding fragments thereof) comprising HCVR and LCVR sequences of Table 4 within a combination of a light chain selected from IgKV4-1, IgKV1-5, IgKV1-9, IgKV1-12, IgKV3-15, IgKV1-16, IgKV1-17, IgKV3-20, IgLV3-21, IgKV2-24, IgKV1-33, IgKV1-39, IgLV1-40, IgLV1-44, IgLV1-51, IgLV3-1, IgKV1-6, IgLV2-8, IgKV3-11, IgLV2-11, IgLV2-14, IgLV2-23, or IgLV6-57, and a heavy chain selected from IgHV1-69, IgHV3-64, IgHV4-59, IgHV3-53, IgHV3-48, IgHV4-34, IgHV3-33, IgHV3-30, IgHV3-23, IgHV3-20, IgHV1-18, IgHV3-15, IgHV3-11, IgHV3-9, IgHV1-8, IgHV3-7, IgHV2-5, IgHV1-2, IgHV2-70, IgHV3-66, IgHV5-51, IgHV1-46, IgHV4-39, IgHV4-31, IgHV3-30-3, IgHV2-26, or IgHV7-4-1.

[0096] Typically, the variable domains of both the heavy and light immunoglobulin chains comprise three hypervariable regions, also called complementarity determining regions (CDRs), located within relatively conserved framework regions (FR). In general, from N-terminal to C-terminal, both light and heavy chains variable domains comprise FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. In an embodiment of the invention, the assignment of amino acids to each domain is in accordance with the definitions of Sequences of Proteins of Immunological Interest, Kabat, et al.; National Institutes of Health, Bethesda, Md.; 5.sup.th ed.; NIH Publ. No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32:1-75; Kabat, et al., (1977) J. Biol. Chem. 252:6609-6616; Chothia, et al., (1987) J Mol. Biol. 196:901-917 or Chothia, et al., (1989) Nature 342:878-883.

[0097] The present invention includes monoclonal anti-CoV-S antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof, as well as monoclonal compositions comprising a plurality of isolated monoclonal antigen-binding proteins. The term monoclonal antibody, as used herein, refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts. A plurality of such monoclonal antibodies and fragments in a composition refers to a concentration of identical (i.e., as discussed above, in amino acid sequence except for possible naturally occurring mutations that may be present in minor amounts) antibodies and fragments which is above that which would normally occur in nature, e.g., in the blood of a host organism such as a mouse or a human.

[0098] In an embodiment of the invention, an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment comprises a heavy chain constant domain, e.g., of the type IgA (e.g., IgA1 or IgA2), IgD, IgE, IgG (e.g., IgG1, IgG2, IgG3 and IgG4) or IgM. In an embodiment of the invention, an antigen-binding protein, e.g., antibody or antigen-binding fragment comprises a light chain constant domain, e.g., of the type kappa or lambda.

[0099] The term human antigen-binding protein, such as an antibody, as used herein, includes antibodies having variable and constant regions derived from human germline immunoglobulin sequences whether in a human cell or grafted into a non-human cell, e.g., a mouse cell. See e.g., U.S. Pat. No. 8,502,018, 6,596,541 or 5,789,215. The human mAbs of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutation in vivo), for example in the CDRs and, in particular, CDR3. However, the term human antibody, as used herein, is not intended to include mAbs in which CDR sequences derived from the germline of another mammalian species (e.g., mouse) have been grafted onto human FR sequences. The term includes antibodies recombinantly produced in a non-human mammal or in cells of a non-human mammal. The term is not intended to include antibodies isolated from or generated in a human subject. See below.

[0100] The present invention includes anti-CoV-S chimeric antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof, and methods of use thereof. As used herein, a chimeric antibody is an antibody having the variable domain from a first antibody and the constant domain from a second antibody, where the first and second antibodies are from different species. (U.S. Pat. No. 4,816,567; and Morrison et al., (1984) Proc. Natl. Acad. Sci. USA 81: 6851-6855).

[0101] The present invention includes anti-CoV-S hybrid antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof, and methods of use thereof. As used herein, a hybrid antibody is an antibody having the variable domain from a first antibody and the constant domain from a second antibody, wherein the first and second antibodies are from different animals, or wherein the variable domain, but not the constant region, is from a first animal. For example, a variable domain can be taken from an antibody isolated from a human and expressed with a fixed constant region not isolated from that antibody. Exemplary hybrid antibodies are described in Example 1, which refers to antibody heavy chain variable region and light chain variable region derived PCR products that were cloned into expression vectors containing a heavy constant region and a light constant region, respectively. Hybrid antibodies are synthetic and non-naturally occurring because the variable and constant regions they contain are not isolated from a single natural source.

[0102] The term recombinant antigen-binding proteins, such as antibodies or antigen-binding fragments thereof, refers to such molecules created, expressed, isolated or obtained by technologies or methods known in the art as recombinant DNA technology which include, e.g., DNA splicing and transgenic expression. The term includes antibodies expressed in a non-human mammal (including transgenic non-human mammals, e.g., transgenic mice), or a cell (e.g., CHO cells) expression system, or a non-human cell expression system, or isolated from a recombinant combinatorial human antibody library. In some embodiments, a recombinant antibody shares a sequence with an antibody isolated from an organism (e.g., a mouse or a human), but has been expressed via recombinant DNA technology. Such antibodies may have post-translational modifications (e.g., glycosylation) that differ from the antibody as isolated from the organism.

[0103] In some embodiments, the antibodies disclosed herein lack fucose in its constant region glycosylation. Methods of measuring fucose in an antibody composition have been described in the art, e.g., U.S. Pat. No. 8,409,838 (Regeneron Pharmaceuticals), incorporated herein by reference. In some embodiments, fucose is undetectable in a composition comprising a population of antibody molecules. In some embodiments, an antibody lacking fucose has enhanced ADCC activity.

[0104] In some embodiments, antibodies that lack fucose can be produced using cell lines that are deficient in their ability to fucosylate proteins, i.e., the ability to fucosylate proteins is reduced or eliminated. Fucosylation of glycans requires synthesis of GDP-fucose via the de novo pathway or the salvage pathway, both of which involve sequential function of several enzymes, leading to addition of a fucose molecule to the first N-acetylglucosamine (GlcNAc) moiety of the reducing end of a glycan. The two key enzymes of the de novo pathway responsible for production of GDP-fucose are GDP-D-mannose-4,6-dehydratase (GMD) and GDP-keto-6-deoxymannose-3,5-epimerase,4-reductase (FX). In the absence of fucose, these two de novo pathway enzymes (GMD and FX) convert mannose and/or glucose to GDP-fucose which is then transported into the Golgi complex where nine fucosyl-transferases (FUT1-9) act in concert to fucosylate the first GlcNAc molecule of a glycan. In the presence of fucose, however, the salvage pathway enzymes, fucose-kinase and GDP-fucose pyrophosphorylase, convert fucose into GDP-fucose.

[0105] Cell lines that are deficient in their ability to fucosylate proteins have been described in the art. In some embodiments, a cell line deficient in its ability to fucosylate proteins is a mammalian cell line (e.g., CHO cell lines, such as CHO K1, DXB-11 CHO, Veggie-CHO) comprising a mutation or genetic modification in one or more of endogenous FUT1 to 9 genes resulting in a lack of one or more functional fucosyl-transferases. In some embodiments, the mammalian cell line comprises a mutation in an endogenous FUT8 gene (e.g., a FUT8 knock-out cell line in which the FUT8 gene has been disrupted resulting in a lack of a functional 1,6-fucosyltransferase in the cell line, as described in U.S. Pat. No. 7,214,775 (Kyowa Hakko Kogyo Co., Ltd.) and U.S. Pat. No. 7,737,725 (Kyowa Hakko Kirin Co., Ltd), incorporated herein by reference. In some embodiments, the mammalian cell line comprises a mutation or genetic modification in an endogenous GMD gene resulting in a lack of a functional GMD in the cell line, e.g., a GMD knock-out cell line in which the GMD gene has been disrupted, described in e.g., U.S. Pat. No. 7,737,725 (Kyowa Hakko Kirin Co., Ltd), incorporated herein by reference. In some embodiments, the mammalian cell line comprises a mutation or genetic modification in an endogenous Fx gene resulting in a lack of a functional Fx protein. In some embodiments, the mammalian cell line is an Fx knock-out cell line in which the endogenous Fx gene has been disrupted (see, e.g., U.S. Pat. No. 7,737,725 (Kyowa Hakko Kirin Co., Ltd), incorporated herein by reference). In some embodiments, the mammalian cell line comprises a mutation in an endogenous Fx mutation that confers temperature sensitive phenotypes (as described in, e.g., U.S. Pat. No. 8,409,838 (Regeneron Pharmaceuticals), incorporated herein by reference). In some embodiments, the mammalian cell line deficient in its ability to fucosylate proteins is a cell line that has been selected based on resistance to certain lectins, e.g., the Lens culinaris lectin. See, e.g., U.S. Pat. No. 8,409,838 (Regeneron Pharmaceuticals), incorporated herein by reference.

[0106] Recombinant anti-CoV-S antigen-binding proteins, e.g., antibodies and antigen-binding fragments, disclosed herein may also be produced in an E. coli/7 expression system. In this embodiment, nucleic acids encoding the anti-CoV-S antibody immunoglobulin molecules of the invention (e.g., as found in Table 4) may be inserted into a pET-based plasmid and expressed in the E. coli/T7 system. For example, the present invention includes methods for expressing an antibody or antigen-binding fragment thereof or immunoglobulin chain thereof in a host cell (e.g., bacterial host cell such as E. coli such as BL21 or BL21DE3) comprising expressing T7 RNA polymerase in the cell which also includes a polynucleotide encoding an immunoglobulin chain that is operably linked to a T7 promoter. For example, in an embodiment of the invention, a bacterial host cell, such as an E. coli, includes a polynucleotide encoding the T7 RNA polymerase gene operably linked to a lac promoter and expression of the polymerase and the chain is induced by incubation of the host cell with IPTG (isopropyl-beta-D-thiogalactopyranoside). See U.S. Pat. Nos. 4,952,496 and 5,693,489 or Studier & Moffatt, Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes, J. Mol. Biol. 1986 May 5; 189(1): 113-30.

[0107] There are several methods by which to produce recombinant antibodies which are known in the art. One example of a method for recombinant production of antibodies is disclosed in U.S. Pat. No. 4,816,567.

[0108] Transformation can be by any known method for introducing polynucleotides (e.g., DNA or RNA, including mRNA) into a host cell. Methods for introduction of heterologous polynucleotides into mammalian cells are well known in the art and include dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of the polynucleotide(s) in liposomes, lipid nanoparticle technology, biolistic injection and direct microinjection of the DNA into nuclei. In addition, nucleic acid molecules may be introduced into mammalian cells by viral vectors such as lentivirus or adeno-associated virus. Methods of transforming cells are well known in the art. See, for example, U.S. Pat. Nos. 4,399,216; 4,912,040; 4,740,461 and 4,959,455. In some embodiments, an antibody or antigen-binding fragment thereof of the present disclosure can be introduced to a subject in nucleic acid form (e.g, DNA or RNA, including mRNA), such that the subject's own cells produce the antibody. The present disclosure further provides modifications to nucleotide sequences encoding the anti-CoV-S antibodies described herein that result in increased antibody expression, increased antibody stability, increased nucleic acid (e.g., mRNA) stability, or improved affinity or specificity of the antibodies for the CoV spike protein.

[0109] Thus, the present invention includes recombinant methods for making an anti-CoV-S antigen-binding protein, such as an antibody or antigen-binding fragment thereof of the present invention, or an immunoglobulin chain thereof, comprising (i) introducing one or more polynucleotides (e.g., including the nucleotide sequence of any one or more of the sequences of Table 5) encoding light and/or heavy immunoglobulin chains, or CDRs, of the antigen-binding protein, e.g., of Table 4, for example, wherein the polynucleotide is in a vector, and/or integrated into a host cell chromosome and/or is operably linked to a promoter, (ii) culturing the host cell (e.g., CHO or Pichia or Pichia pastoris) under condition favorable to expression of the polynucleotide and, (iii) optionally, isolating the antigen-binding protein, (e.g., antibody or fragment) or chain from the host cell and/or medium in which the host cell is grown. For example, a polynucleotide can be integrated into a host cell chromosome through targeted insertion with a vector such as adeno-associated virus (AAV), e.g., after cleavage of the chromosome using a gene editing system (e.g., CRISPR (for example, CRISPR-Cas9), TALEN, megaTAL, zinc finger, or Argonaute). Targeted insertions can take place, for example, at host cell loci such as an albumin or immunoglopbulin genomic locus. Alternatively, insertion can be at a random locus, e.g., using a vector such as lentivirus. When making an antigen-binding protein (e.g., antibody or antigen-binding fragment) comprising more than one immunoglobulin chain, e.g., an antibody that comprises two heavy immunoglobulin chains and two light immunoglobulin chains, co-expression of the chains in a single host cell leads to association of the chains, e.g., in the cell or on the cell surface or outside the cell if such chains are secreted, so as to form the antigen-binding protein (e.g., antibody or antigen-binding fragment). The methods include those wherein only a heavy immunoglobulin chain or only a light immunoglobulin chain (e.g., any of those discussed herein including mature fragments and/or variable domains thereof) is expressed. Such chains are useful, for example, as intermediates in the expression of an antibody or antigen-binding fragment that includes such a chain. For example, the present invention also includes anti-CoV-S antigen-binding proteins, such as antibodies and antigen-binding fragments thereof, comprising a heavy chain immunoglobulin (or variable domain thereof or comprising the CDRs thereof) encoded by a polynucleotide comprising a nucleotide sequence set forth in Table 5 and a light chain immunoglobulin (or variable domain thereof or comprising the CDRs thereof) encoded by a nucleotide sequence set forth in Table 5 which are the product of such production methods, and, optionally, the purification methods set forth herein. For example, in some embodiments, the product of the method is an anti-CoV-S antigen-binding protein which is an antibody or fragment comprising an HCVR comprising an amino acid sequence set forth in Table 4 and an LCVR comprising an amino acid sequence set forth in Table 4, wherein the HCVR and LCVR sequences are selected from a single antibody listed in Table 4. In some embodiments, the product of the method is an anti-CoV-S antigen-binding protein which is an antibody or fragment comprising HCDR1, HCDR2, and HCDR3 comprising amino acid sequences set forth in Table 4 and LCDR1, LCDR2, and LCDR3 comprising amino acid sequences set forth in Table 4, wherein the six CDR sequences are selected from a single antibody listed in Table 4. In some embodiments, the product of the method is an anti-CoV-S antigen-binding protein which is an antibody or fragment comprising a heavy chain comprising an HC amino acid sequence set forth in Table 4 and a light chain comprising an LC amino acid sequence set forth in Table 4.

[0110] Eukaryotic and prokaryotic host cells, including mammalian cells, may be used as hosts for expression of an anti-CoV-S antigen-binding protein. Such host cells are well known in the art and many are available from the American Type Culture Collection (ATCC). These host cells include, inter alia, Chinese hamster ovary (CHO) cells, NS0, SP2 cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, 3T3 cells, HEK-293 cells and a number of other cell lines. Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, bovine, horse and hamster cells. Other cell lines that may be used are insect cell lines (e.g., Spodoptera frugiperda or Trichoplusia ni), amphibian cells, bacterial cells, plant cells and fungal cells. Fungal cells include yeast and filamentous fungus cells including, for example, Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichia minuta (Ogataea minuta, Pichia lindneri), Pichia opuntiae, Pichia thermotolerans, Pichia salictaria, Pichia guercuum, Pichia pijperi, Pichia stiptis, Pichia methanolica, Pichia sp., Saccharomyces cerevisiae, Saccharomyces sp., Hansenula polymorpha, Kluyveromyces sp., Kluyveromyces lactis, Candida albicans, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium lucknowense, Fusarium sp., Fusarium gramineum, Fusarium venenatum, Physcomitrella patens and Neurospora crassa. The present invention includes an isolated host cell (e.g., a CHO cell) comprising an antigen-binding protein, such as those of Table 4; or a polynucleotide encoding such a polypeptide thereof.

[0111] Polynucleotides, as discussed herein, may encode all or a portion of an antibody or antigen-binding fragment as discussed throughout the present disclosure. In some cases, a single polynucleotide may encode both a HCVR and a LCVR (e.g., defined with reference to the CDRs contained within the respective amino acid sequence-defined HCVR and LCVR, defined with reference to the amino acid sequences of the CDRs of the HCVR and LCVR, respectively, or defined with reference to the amino acid sequences of the HCVR and LCVR, respectively) of an antibody or antigen-binding fragment, or the HCVR and LCVR may be encoded by separate polynucleotides (i.e., a pair of polynucleotides). In the latter case, in which the HCVR and LCVR are encoded by separate polynucleotides, the polynucleotides may be combined in a single vector or may be contained in separate vectors (i.e., a pair of vectors). In any case, a host cell used to express the polynucleotide(s) or vector(s) may contain the full complement of component parts to generate the antibody or antigen-binding fragment thereof. For example, a host cell may comprise separate vectors, each encoding a HCVR and a LCVR, respectively, of an antibody or antigen-binding fragment thereof as discussed above or herein. Similarly, the polynucleotide or polynucleotides, and the vector or vectors, may be used to express the full-length heavy chain and full-length light chain of an antibody as discussed above or herein. For example, a host cell may comprise a single vector with polynucleotides encoding both a heavy chain and a light chain of an antibody, or the host cell may comprise separate vectors with polynucleotides encoding, respectively, a heavy chain and a light chain of an antibody as discussed above or herein.

[0112] In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the HCDRs of SEQ ID NOs: 214, 216 and 218 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 213, 215 and 217). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the LCDRs of SEQ ID NOs: 222, 126 and 224 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 221, 125 and 223). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the amino acid sequence of SEQ ID NO: 212 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 211). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the amino acid sequence of SEQ ID NO: 220 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 219). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 226 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 225). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 228 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 227). In an embodiment, the present disclosure provides a pair of polynucleotides or a pair of vectors encoding, respectively, a HCVR and a LCVR, or a HC and a LC, as discussed in this paragraph, as well as a host cell containing the pair or polynucleotides and/or the pair of vectors to encode the antibody designated mAb14315 or an antigen-binding fragment thereof.

[0113] In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the HCDRs of SEQ ID NOs: 364, 366 and 368 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 363, 365 and 367). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the LCDRs of SEQ ID NOs: 372, 106 and 374 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 371, 105 and 373). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the amino acid sequence of SEQ ID NO: 362 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 361). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the amino acid sequence of SEQ ID NO: 370 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 369). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 376 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 375). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 378 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 377). In an embodiment, the present disclosure provides a pair of polynucleotides or a pair of vectors encoding, respectively, a HCVR and a LCVR, or a HC and a LC, as discussed in this paragraph, as well as a host cell containing the pair or polynucleotides and/or the pair of vectors to encode the antibody designated mAb15160 or an antigen-binding fragment thereof.

[0114] In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the HCDRs of SEQ ID NOs: 495, 497 and 499 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 494, 496 and 498). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the LCDRs of SEQ ID NOs: 503, 505 and 507 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 502, 504 and 506). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the amino acid sequence of SEQ ID NO: 493 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 492). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the amino acid sequence of SEQ ID NO: 501 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 500). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 509 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 508). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 511 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 510). In an embodiment, the present disclosure provides a pair of polynucleotides or a pair of vectors encoding, respectively, a HCVR and a LCVR, or a HC and a LC, as discussed in this paragraph, as well as a host cell containing the pair or polynucleotides and/or the pair of vectors to encode the antibody designated mAb14284 or an antigen-binding fragment thereof.

[0115] In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the HCDRs of SEQ ID NOs: 889, 891 and 893 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 888, 890 and 892). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the LCDRs of SEQ ID NOs: 897, 164 and 899 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 896, 163 and 898). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the amino acid sequence of SEQ ID NO: 887 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 886). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the amino acid sequence of SEQ ID NO: 895 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 894). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 901 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 900). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 903 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 902). In an embodiment, the present disclosure provides a pair of polynucleotides or a pair of vectors encoding, respectively, a HCVR and a LCVR, or a HC and a LC, as discussed in this paragraph, as well as a host cell containing the pair or polynucleotides and/or the pair of vectors to encode the antibody designated mAb14256 or an antigen-binding fragment thereof.

[0116] In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the HCDRs of SEQ ID NOs: 495, 497 and 499 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 494, 496 and 498). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the LCDRs of SEQ ID NOs: 503, 505 and 507 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 502, 504 and 506). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the amino acid sequence of SEQ ID NO: 493 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 492). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the amino acid sequence of SEQ ID NO: 501 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 500). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 1075 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 1074). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 511 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 510). In an embodiment, the present disclosure provides a pair of polynucleotides or a pair of vectors encoding, respectively, a HCVR and a LCVR, or a HC and a LC, as discussed in this paragraph, as well as a host cell containing the pair or polynucleotides and/or the pair of vectors to encode the antibody designated mAb17090 or an antigen-binding fragment thereof.

[0117] In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the HCDRs of SEQ ID NOs: 364, 366 and 368 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 363, 365 and 367). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the LCDRs of SEQ ID NOs: 372, 106 and 374 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NOs: 371, 105 and 373). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a HCVR comprising the amino acid sequence of SEQ ID NO: 362 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 361). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a LCVR comprising the amino acid sequence of SEQ ID NO: 370 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 369). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a heavy chain (HC) comprising the amino acid sequence of SEQ ID NO: 1077 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 1076). In an exemplary embodiment, the present disclosure provides a polynucleotide (or a vector comprising the polynucleotide, or a host cell comprising the polynucleotide or the vector) encoding a light chain (LC) comprising the amino acid sequence of SEQ ID NO: 378 (e.g., in an embodiment, the polynucleotide comprises SEQ ID NO: 377). In an embodiment, the present disclosure provides a pair of polynucleotides or a pair of vectors encoding, respectively, a HCVR and a LCVR, or a HC and a LC, as discussed in this paragraph, as well as a host cell containing the pair or polynucleotides and/or the pair of vectors to encode the antibody designated mAb15160_2 or an antigen-binding fragment thereof.

[0118] The term specifically binds refers to those antigen-binding proteins (e.g., mAbs) having a binding affinity to an antigen, such as a CoV-S protein (e.g., SARS-CoV-2-S), expressed as K.sub.D, of at least about 10.sup.8 M, as measured by real-time, label free bio-layer interferometry assay, for example, at 25 C. or 37 C., e.g., an Octet HTX biosensor, or by surface plasmon resonance, e.g., BIACORE, or by solution-affinity ELISA. The present invention includes antigen-binding proteins that specifically bind to a CoV-S protein.

[0119] The terms antigen-binding portion or antigen-binding fragment of an antibody or antigen-binding protein, and the like, as used herein, include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds an antigen to form a complex. Non-limiting examples of antigen-binding fragments include: (i) Fab fragments; (ii) F(ab).sub.2 fragments; (iii) Fd fragments; (iv) Fv fragments; (v) single-chain Fv (scFv) molecules; (vi) dAb fragments; and (vii) minimal recognition units consisting of the amino acid residues that mimic the hypervariable region of an antibody (e.g., an isolated complementarity determining region (CDR) such as a CDR3 peptide), or a constrained FR3-CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., as defined in WO08/020079 or WO09/138519) (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed within the expression antigen-binding fragment, as used herein. In an embodiment of the invention, the antigen-binding fragment comprises three or more CDRs of an antibody of Table 4 (e.g., HCDR1, HCDR2 and HCDR3; or LCDR1, LCDR2 and LCDR3).

[0120] An antigen-binding fragment of an antibody will, in an embodiment of the invention, comprise at least one variable domain. The variable domain may be of any size or amino acid composition and will generally comprise at least one CDR, which is adjacent to or in frame with one or more framework sequences. In antigen-binding fragments having a V.sub.H domain associated with a V.sub.L domain, the V.sub.H and V.sub.L domains may be situated relative to one another in any suitable arrangement. For example, the variable region may be dimeric and contain V.sub.H-V.sub.H, V.sub.H-V.sub.L or V.sub.L-V.sub.L dimers. Alternatively, the antigen-binding fragment of an antibody may contain a monomeric V.sub.H or V.sub.L domain.

[0121] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting, exemplary configurations of variable and constant domains that may be found within an antigen-binding fragment of an antibody of the present invention include: (i) V.sub.H-C.sub.H1; (ii) V.sub.H-C.sub.H2; (iii) V.sub.H-C.sub.H3; (iv) V.sub.HC.sub.H1-C.sub.H2; (v) V.sub.H-C.sub.H1-C.sub.H2-C.sub.H3; (vi) V.sub.H-C.sub.H2-C.sub.H3; (vii) V.sub.H-C.sub.L; (viii) V.sub.L-C.sub.H1; (ix) V.sub.L-C.sub.H2; (x) V.sub.L-C.sub.H3; (xi) V.sub.L-C.sub.H1-C.sub.H2; (xii) V.sub.L-C.sub.H1-C.sub.H2-C.sub.H3; (xiii) V.sub.L-C.sub.H2-C.sub.H3; and (xiv) V.sub.L-C.sub.L. In any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be either directly linked to one another or may be linked by a full or partial hinge or linker region. A hinge region may consist of at least 2 (e.g., 5, 10, 15, 20, 40, 60 or more) amino acids, which result in a flexible or semi-flexible linkage between adjacent variable and/or constant domains in a single polypeptide molecule. Moreover, an antigen-binding fragment of an antibody of the present invention may comprise a homo-dimer or hetero-dimer (or other multimer) of any of the variable and constant domain configurations listed above in non-covalent association with one another and/or with one or more monomeric V.sub.H or V.sub.L domain (e.g., by disulfide bond(s)).

[0122] Antigen-binding proteins (e.g., antibodies and antigen-binding fragments) may be mono-specific or multi-specific (e.g., bi-specific). Multispecific antigen-binding proteins are discussed further herein.

[0123] In specific embodiments, antibody or antibody fragments of the invention may be conjugated to a moiety such a ligand or a therapeutic moiety (immunoconjugate), such as an anti-viral drug, a second anti-influenza antibody, or any other therapeutic moiety useful for treating a viral infection, e.g., influenza viral infection. See below.

[0124] The present invention also provides a complex comprising an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment, discussed herein complexed with CoV-S polypeptide or an antigenic fragment thereof and/or with a secondary antibody or antigen-binding fragment thereof (e.g., detectably labeled secondary antibody) that binds specifically to the anti-CoV-S antibody or fragment. In an embodiment of the invention, the antibody or fragment is in vitro (e.g., is immobilized to a solid substrate) or is in the body of a subject. In an embodiment of the invention, the CoV-S is in vitro (e.g., is immobilized to a solid substrate) or is on the surface of a virus or is in the body of a subject. Immobilized anti-CoV-S antibodies and antigen-binding fragments thereof which are covalently linked to an insoluble matrix material (e.g., glass or polysaccharide such as agarose or sepharose, e.g., a bead or other particle thereof) are also part of the present invention; optionally, wherein the immobilized antibody is complexed with CoV-S or antigenic fragment thereof or a secondary antibody or fragment thereof.

[0125] Isolated antigen-binding proteins, antibodies or antigen-binding fragments thereof, polypeptides, polynucleotides and vectors, are at least partially free of other biological molecules from the cells or cell culture from which they are produced. Such biological molecules include nucleic acids, proteins, other antibodies or antigen-binding fragments, lipids, carbohydrates, or other material such as cellular debris and growth medium. An isolated antibody or antigen-binding fragment may further be at least partially free of expression system components such as biological molecules from a host cell or of the growth medium thereof. Generally, the term isolated is not intended to refer to a complete absence of such biological molecules or to an absence of water, buffers, or salts or to components of a pharmaceutical formulation that includes the antibodies or fragments.

[0126] The term epitope refers to an antigenic determinant (e.g., a CoV-S polypeptide) that interacts with a specific antigen-binding site of an antigen-binding protein, e.g., a variable region of an antibody molecule, known as a paratope. A single antigen may have more than one epitope. Thus, different antibodies may bind to different areas on an antigen and may have different biological effects. The term epitope also refers to a site on an antigen to which B and/or T cells respond. It also refers to a region of an antigen that is bound by an antibody. Epitopes may be defined as structural or functional. Functional epitopes are generally a subset of the structural epitopes and have those residues that directly contribute to the affinity of the interaction. Epitopes may be linear or conformational, that is, composed of non-linear amino acids. In certain embodiments, epitopes may include determinants that are chemically active surface groupings of molecules such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and, in certain embodiments, may have specific three-dimensional structural characteristics, and/or specific charge characteristics.

[0127] Methods for determining the epitope of an antigen-binding protein, e.g., antibody or fragment or polypeptide, include alanine scanning mutational analysis, peptide blot analysis (Reineke (2004) Methods Mol. Biol. 248: 443-63), peptide cleavage analysis, crystallographic studies and NMR analysis. In addition, methods such as epitope excision, epitope extraction and chemical modification of antigens can be employed (Tomer (2000) Prot. Sci. 9: 487-496). Another method that can be used to identify the amino acids within a polypeptide with which an antigen-binding protein (e.g., antibody or fragment or polypeptide) (e.g., coversin) interacts is hydrogen/deuterium exchange detected by mass spectrometry. In general terms, the hydrogen/deuterium exchange method involves deuterium-labeling the protein of interest, followed by binding the antigen-binding protein, e.g., antibody or fragment or polypeptide, to the deuterium-labeled protein. Next, the CoV-S protein/antigen-binding protein complex is transferred to water and exchangeable protons within amino acids that are protected by the antibody complex undergo deuterium-to-hydrogen back-exchange at a slower rate than exchangeable protons within amino acids that are not part of the interface. As a result, amino acids that form part of the protein/antigen-binding protein interface may retain deuterium and therefore exhibit relatively higher mass compared to amino acids not included in the interface. After dissociation of the antigen-binding protein (e.g., antibody or fragment or polypeptide), the target protein is subjected to protease cleavage and mass spectrometry analysis, thereby revealing the deuterium-labeled residues which correspond to the specific amino acids with which the antigen-binding protein interacts. See, e.g., Ehring (1999) Analytical Biochemistry 267: 252-259; Engen and Smith (2001) Anal. Chem. 73: 256A-265A.

[0128] The term competes as used herein, refers to an antigen-binding protein (e.g., antibody or antigen-binding fragment thereof) that binds to an antigen (e.g., CoV-S) and inhibits or blocks the binding of another antigen-binding protein (e.g., antibody or antigen-binding fragment thereof) to the antigen. The term also includes competition between two antigen-binding proteins e.g., antibodies, in both orientations, i.e., a first antibody that binds and blocks binding of second antibody and vice versa. In certain embodiments, the first antigen-binding protein (e.g., antibody) and second antigen-binding protein (e.g., antibody) may bind to the same epitope. Alternatively, the first and second antigen-binding proteins (e.g., antibodies) may bind to different, but, for example, overlapping epitopes, wherein binding of one inhibits or blocks the binding of the second antibody, e.g., via steric hindrance. Competition between antigen-binding proteins (e.g., antibodies) may be measured by methods known in the art, for example, by a real-time, label-free bio-layer interferometry assay. Epitope mapping (e.g., via alanine scanning or hydrogen-deuterium exchange (HDX)) can be used to determine whether two or more antibodies are non-competing (e.g., on a spike protein receptor binding domain (RBD) monomer), competing for the same epitope, or competing but with diverse micro-epitopes (e.g., identified through HDX). In an embodiment of the invention, competition between a first and second anti-CoV-S antigen-binding protein (e.g., antibody) is determined by measuring the ability of an immobilized first anti-CoV-S antigen-binding protein (e.g., antibody) (not initially complexed with CoV-S protein) to bind to soluble CoV-S protein complexed with a second anti-CoV-S antigen-binding protein (e.g., antibody). A reduction in the ability of the first anti-CoV-S antigen-binding protein (e.g., antibody) to bind to the complexed CoV-S protein, relative to uncomplexed CoV-S protein, indicates that the first and second anti-CoV-S antigen-binding proteins (e.g., antibodies) compete. The degree of competition can be expressed as a percentage of the reduction in binding. Such competition can be measured using a real time, label-free bio-layer interferometry assay, e.g., on an Octet RED384 biosensor (Pall ForteBio Corp.), ELISA (enzyme-linked immunosorbent assays) or SPR (surface plasmon resonance).

[0129] Binding competition between anti-CoV-S antigen-binding proteins (e.g., monoclonal antibodies (mAbs)) can be determined using a real time, label-free bio-layer interferometry assay on an Octet RED384 biosensor (Pall ForteBio Corp.). For example, to determine competition between two anti-CoV-S monoclonal antibodies, the anti-CoV-S mAb can be first captured onto anti-hFc antibody coated Octet biosensor tips (Pall ForteBio Corp., #18-5060) by submerging the tips into a solution of anti-CoV-S mAb (subsequently referred to as mAb1). As a positive-control for blocking, the antibody captured biosensor tips can then be saturated with a known blocking isotype control mAb (subsequently referred to as blocking mAb) by dipping into a solution of blocking mAb. To determine if mAb2 competes with mAb1, the biosensor tips can then be subsequently dipped into a co-complexed solution of CoV-S polypeptide and a second anti-CoV-S mAb (subsequently referred to as mAb2), that had been pre-incubated for a period of time and binding of mAb1 to the CoV-S polypeptide can be determined. The biosensor tips can be washed in buffer in between every step of the experiment. The real-time binding response can be monitored during the course of the experiment and the binding response at the end of every step can be recorded.

[0130] For example, in an embodiment of the invention, the competition assay is conducted at 25 C. and pH about 7, e.g., 7.4, e.g., in the presence of buffer, salt, surfactant and a non-specific protein (e.g., bovine serum albumin).

[0131] Typically, an antibody or antigen-binding fragment of the invention which is modified in some way retains the ability to specifically bind to CoV-S, e.g., retains at least 10% of its CoV-S binding activity (when compared to the parental antibody) when that activity is expressed on a molar basis. Preferably, an antibody or antigen-binding fragment of the invention retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or more of the CoV-S binding affinity as the parental antibody. It is also intended that an antibody or antigen-binding fragment of the invention can include conservative or non-conservative amino acid substitutions (referred to as conservative variants or function conserved variants of the antibody) that do not substantially alter its biologic activity.

[0132] A variant of a polypeptide, such as an immunoglobulin chain (e.g., mAb8021 V.sub.H, V.sub.L, HC, or LC, mAb8028 V.sub.H, V.sub.L, HC, or LC, or mAb8029 V.sub.H, V.sub.L, HC, or LC), refers to a polypeptide comprising an amino acid sequence that is at least about 70-99.9% (e.g., 70, 72, 74, 75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, 99.9%) identical or similar to a referenced amino acid sequence that is set forth herein (e.g., SEQ ID NO: 2, 10, 18, 20, 22, 30, 38, 40, 42, 50, 58, or 60); when the comparison is performed by a BLAST algorithm wherein the parameters of the algorithm are selected to give the largest match between the respective sequences over the entire length of the respective reference sequences (e.g., expect threshold: 10; word size: 3; max matches in a query range: 0; BLOSUM 62 matrix; gap costs: existence 11, extension 1; conditional compositional score matrix adjustment).

[0133] A variant of a polynucleotide refers to a polynucleotide comprising a nucleotide sequence that is at least about 70-99.9% (e.g., at least about 70, 72, 74, 75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, or 99.9%) identical to a referenced nucleotide sequence that is set forth herein (e.g., SEQ ID NO: 1, 9, 17, 19, 21, 29, 37, 39, 41, 49, 57, or 59); when the comparison is performed by a BLAST algorithm wherein the parameters of the algorithm are selected to give the largest match between the respective sequences over the entire length of the respective reference sequences (e.g., expect threshold: 10; word size: 28; max matches in a query range: 0; match/mismatch scores: 1, 2; gap costs: linear).

[0134] Anti-CoV-S antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof of the present invention, in an embodiment of the invention, include a heavy chain immunoglobulin variable region having at least 70% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater) amino acid sequence identity to the HCVR amino acid sequences set forth in Table 4; and/or a light chain immunoglobulin variable region having at least 70% (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater) amino acid sequence identity to the LCVR amino acid sequences set forth in Table 4.

[0135] In addition, a variant anti-CoV-S antigen-binding protein may include a polypeptide comprising an amino acid sequence that is set forth herein except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10) mutations such as, for example, missense mutations (e.g., conservative substitutions), non-sense mutations, deletions, or insertions. For example, the present invention includes antigen-binding proteins which include an immunoglobulin light chain variant comprising an LCVR amino acid sequence set forth in Table 4 but having one or more of such mutations and/or an immunoglobulin heavy chain variant comprising an HCVR amino acid sequence set forth in Table 4 but having one or more of such mutations. In an embodiment of the invention, a variant anti-CoV-S antigen-binding protein includes an immunoglobulin light chain variant comprising LCDR1, LCDR2 and LCDR3 wherein one or more (e.g., 1 or 2 or 3) of such CDRs has one or more of such mutations (e.g., conservative substitutions) and/or an immunoglobulin heavy chain variant comprising HCDR1, HCDR2 and HCDR3 wherein one or more (e.g., 1 or 2 or 3) of such CDRs has one or more of such mutations (e.g., conservative substitutions). Substitutions can be in a CDR, framework, or constant region.

[0136] The invention further provides variant anti-CoV-S antigen-binding proteins, e.g., antibodies or antigen-binding fragments thereof, comprising one or more variant CDRs (e.g., any one or more of LCDR1, LCDR2, LCDR3, HCDR1, HCDR2 and/or HCDR3) that are set forth herein with at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 99.9% sequence identity or similarity to, e.g., the heavy chain and light chain CDRs of Table 4.

[0137] Embodiments of the present invention also include variant antigen-binding proteins, e.g., anti-CoV-S antibodies and antigen-binding fragments thereof, that comprise immunoglobulin V.sub.Hs and V.sub.Ls; or HCs and LCs, which comprise an amino acid sequence having 70% or more (e.g., 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or greater) overall amino acid sequence identity or similarity to the amino acid sequences of the corresponding V.sub.Hs, V.sub.Ls, HCs or LCs specifically set forth herein, but wherein the LCDR1, LCDR2, LCDR3, HCDR1, HCDR2 and HCDR3 of such immunoglobulins are not variants and comprise CDR amino acid sequence set forth in Table 4. Thus, in such embodiments, the CDRs within variant antigen-binding proteins are not, themselves, variants.

[0138] Conservatively modified variant anti-CoV-S antibodies and antigen-binding fragments thereof are also part of the present invention. A conservatively modified variant or a conservative substitution refers to a variant wherein there is one or more substitutions of amino acids in a polypeptide with other amino acids having similar characteristics (e.g. charge, side-chain size, hydrophobicity/hydrophilicity, backbone conformation and rigidity, etc.). Such changes can frequently be made without significantly disrupting the biological activity of the antibody or fragment. Those of skill in this art recognize that, in general, single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin/Cummings Pub. Co., p. 224 (4.sup.th Ed.)). In addition, substitutions of structurally or functionally similar amino acids are less likely to significantly disrupt biological activity.

[0139] Examples of groups of amino acids that have side chains with similar chemical properties include 1) aliphatic side chains: glycine, alanine, valine, leucine and isoleucine; 2) aliphatic-hydroxyl side chains: serine and threonine; 3) amide-containing side chains: asparagine and glutamine; 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan; 5) basic side chains: lysine, arginine, and histidine; 6) acidic side chains: aspartate and glutamate, and 7) sulfur-containing side chains: cysteine and methionine. Preferred conservative amino acids substitution groups are: valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamate-aspartate, and asparagine-glutamine. Alternatively, a conservative replacement is any change having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256: 1443 45.

[0140] Function-conservative variants of the anti-CoV-S antibodies and antigen-binding fragments thereof are also part of the present invention. Any of the variants of the anti-CoV-S antibodies and antigen-binding fragments thereof (as discussed herein) may be function-conservative variants. Such function-conservative variants may, in some cases, also be characterized as conservatively modified variants. Function-conservative variants, as used herein, refers to variants of the anti-CoV-S antibodies or antigen-binding fragments thereof in which one or more amino acid residues have been changed without significantly altering one or more functional properties of the antibody or fragment. In an embodiment of the invention, a function-conservative variant anti-CoV-S antibody or antigen-binding fragment thereof of the present invention comprises a variant amino acid sequence and exhibits one or more of the following functional properties: [0141] Inhibits growth of coronavirus (e.g., SARS-CoV-2, SARS-CoV, and/or MERS-CoV) in ACE2- and/or TMPRSS2-expressing cells (e.g., Calu-3 cells); [0142] Does not significantly bind to MDCK/Tet-on cells which do not express ACE2 and/or TMPRSS2; [0143] Limits spread of coronavirus infection (e.g., by SARS-CoV-2, SARS-CoV, and/or MERS-CoV) of cells, e.g., Calu-3, in vitro; and/or [0144] Protects a mouse engineered to express the human TMPRSS2 and/or ACE2 protein from death caused by coronavirus infection (e.g., SARS-CoV-2, SARS-CoV, or MERS-CoV), for example, wherein the mice are infected with an otherwise lethal dose of the virus, optionally when combined with a second therapeutic agent. [0145] Protects a mouse engineered to express the human TMPRSS2 and/or ACE2 protein from weight loss caused by coronavirus infection (e.g., SARS-CoV-2, SARS-CoV, or MERS-CoV), for example, wherein the mice are infected with a dose of the virus that would otherwise cause weight loss, optionally when combined with a second therapeutic agent.

[0146] A neutralizing or antagonist anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment, refers to a molecule that inhibits an activity of CoV-S to any detectable degree, e.g., inhibits the ability of CoV-S to bind to a receptor such as ACE2, to be cleaved by a protease such as TMPRSS2, or to mediate viral entry into a host cell or viral reproduction in a host cell.

[0147] Table 4 refers to antigen-binding proteins, such as antibodies and antigen-binding fragments thereof, that comprise the heavy chain or V.sub.H (or a variant thereof) and light chain or V.sub.L (or a variant thereof) as set forth below; or that comprise a V.sub.H that comprises the CDRs thereof (HCDR1 (or a variant thereof), HCDR2 (or a variant thereof) and HCDR3 (or a variant thereof)) and a V.sub.L that comprises the CDRs thereof (LCDR1 (or a variant thereof), LCDR2 (or a variant thereof) and LCDR3 (or a variant thereof)), e.g., wherein the immunoglobulin chains, variable regions and/or CDRs comprise the specific amino acid sequences described below.

[0148] The antibodies described herein also include embodiments wherein the V.sub.H is fused to a wild-type IgG4 (e.g., wherein residue 108 is S) or to IgG4 variants (e.g., wherein residue 108 is P).

[0149] Antibodies and antigen-binding fragments of the present invention comprise immunoglobulin chains including the amino acid sequences set forth herein as well as cellular and in vitro post-translational modifications to the antibody. For example, the present invention includes antibodies and antigen-binding fragments thereof that specifically bind to CoV-S comprising heavy and/or light chain amino acid sequences set forth herein (e.g., HCDR1, HCDR2, HCDR3, LCDR1, LCDR2 and/or LCDR3) as well as antibodies and fragments wherein one or more amino acid residues is glycosylated, one or more Asn residues is deamidated, one or more residues (e.g., Met, Trp and/or His) is oxidized, the N-terminal Gln is pyroglutamate (pyroE) and/or the C-terminal Lysine is missing. The amino acid and nucleotide sequences of exemplary anti-SARS-CoV-2-Spike protein (SARS-CoV-2-S) antibodies are shown in the Table of Exemplary Sequences (Table 1), below.

TABLE-US-00001 TABLE1 TableofExemplarySequences Antibody Component Designation Part Sequence SEQIDNO mAb14256 AminoAcids HCVR QVQLQESGPGLVKPSETLSLTCTVSGGSISSHYW 887 SWIRQPPGKGLEWIGYIYYSGSSNYNPSLKSRVT ISVDTSKNQFSLKLNSVTAADTAVYYCARHYDIL TGFDWFDPWGQGTLVTVSS HCDR1 GGSISSHY 889 HCDR2 IYYSGSS 891 HCDR3 ARHYDILTGFDWFDP 893 LCVR QSVLTQPPSVSGAPGQRVTISCTGSSSNIGTHYD 895 VHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSK SGTSASLAITGLQAEDEADYYCQSFDNSLTAPYV FGTGTKVTVL LCDR1 SSNIGTHYD 897 LCDR2 GNS 164 LCDR3 QSFDNSLTAPYV 899 HC QVQLQESGPGLVKPSETLSLTCTVSGGSISSHYW 901 SWIRQPPGKGLEWIGYIYYSGSSNYNPSLKSRVT ISVDTSKNQFSLKLNSVTAADTAVYYCARHYDIL TGFDWFDPWGQGTLVTVSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEL LGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGK LC QSVLTQPPSVSGAPGQRVTISCTGSSSNIGTHYD 903 VHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSK SGTSASLAITGLQAEDEADYYCQSFDNSLTAPYV FGTGTKVTVLGQPKAAPSVTLFPPSSEELQANKA TLVCLISDFYPGAVTVAWKADSSPVKAGVETTTP SKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECS NucleicAcids HCVR caggtgcagctgcaggagtcgggcccaggactgg 886 tgaagccttcggagaccctgtccctcacctgcac tgtctctggtggctccatcagtagtcactactgg agctggatccggcagcccccagggaagggactgg aatggattgggtatatttattacagcgggagctc caactacaacccctccctcaagagtcgagtcacc atatcagtagacacgtccaagaaccagttctccc tgaaactgaattctgtgaccgccgcagacacggc cgtgtattactgtgcgagacattacgatattttg actggttttgactggttcgacccctggggccagg gaaccctggtcaccgtctcctca HCDR1 ggtggctccatcagtagtcactac 888 HCDR2 atttattacagcgggagctcc 890 HCDR3 gcgagacattacgatattttgactggttttgact 892 ggttcgacccc LCVR cagtctgtgctgacgcagccgccctcagtgtcag 894 gggccccagggcagagggtcaccatctcctgcac tgggagcagttccaacatcgggacacattatgat gtacactggtaccaacaacttccaggaacagccc ccaaactcctcatctatggtaacagcaatcggcc ctcaggggtccctgaccgattctctggctccaag tctggcacctcagcctccctggccatcactgggc tccaggctgaggatgaggctgattattactgcca gtcctttgacaacagcctgactgccccttatgtc ttcggaactgggaccaaggtcaccgtccta LCDR1 agttccaacatcgggacacattatgat 896 LCDR2 ggtaacagc 163 LCDR3 cagtcctttgacaacagcctgactgccccttatg 898 tc HC caggtgcagctgcaggagtcgggcccaggactgg 900 tgaagccttcggagaccctgtccctcacctgcac tgtctctggtggctccatcagtagtcactactgg agctggatccggcagcccccagggaagggactgg aatggattgggtatatttattacagcgggagctc caactacaacccctccctcaagagtcgagtcacc atatcagtagacacgtccaagaaccagttctccc tgaaactgaattctgtgaccgccgcagacacggc cgtgtattactgtgcgagacattacgatattttg actggttttgactggttcgacccctggggccagg gaaccctggtcaccgtctcctcagcctccaccaa gggcccatcggtcttccccctggcaccctcctcc aagagcacctctgggggcacagcggccctgggct gcctggtcaaggactacttccccgaaccggtgac ggtgtcgtggaactcaggcgccctgaccagcggc gtgcacaccttcccggctgtcctacagtcctcag gactctactccctcagcagcgtggtgaccgtgcc ctccagcagcttgggcacccagacctacatctgc aacgtgaatcacaagcccagcaacaccaaggtgg acaagaaagttgagcccaaatcttgtgacaaaac tcacacatgcccaccgtgcccagcacctgaactc ctggggggaccgtcagtcttcctcttccccccaa aacccaaggacaccctcatgatctcccggacccc tgaggtcacatgcgtggtggtggacgtgagccac gaagaccctgaggtcaagttcaactggtacgtgg acggcgtggaggtgcataatgccaagacaaagcc gcgggaggagcagtacaacagcacgtaccgtgtg gtcagcgtcctcaccgtcctgcaccaggactggc tgaatggcaaggagtacaagtgcaaggtctccaa caaagccctcccagcccccatcgagaaaaccatc tccaaagccaaagggcagccccgagaaccacagg tgtacaccctgcccccatcccgggatgagctgac caagaaccaggtcagcctgacctgcctggtcaaa ggcttctatcccagcgacatcgccgtggagtggg agagcaatgggcagccggagaacaactacaagac cacgcctcccgtgctggactccgacggctccttc ttcctctacagcaagctcaccgtggacaagagca ggtggcagcaggggaacgtcttctcatgctccgt gatgcatgaggctctgcacaaccactacacgcag aagtccctctccctgtctccgggtaaatga LC cagtctgtgctgacgcagccgccctcagtgtcag 902 gggccccagggcagagggtcaccatctcctgcac tgggagcagttccaacatcgggacacattatgat gtacactggtaccaacaacttccaggaacagccc ccaaactcctcatctatggtaacagcaatcggcc ctcaggggtccctgaccgattctctggctccaag tctggcacctcagcctccctggccatcactgggc tccaggctgaggatgaggctgattattactgcca gtcctttgacaacagcctgactgccccttatgtc ttcggaactgggaccaaggtcaccgtcctaggcc agcccaaggccgccccctccgtgaccctgttccc cccctcctccgaggagctgcaggccaacaaggcc accctggtgtgcctgatctccgacttctaccccg gcgccgtgaccgtggcctggaaggccgactcctc ccccgtgaaggccggcgtggagaccaccaccccc tccaagcagtccaacaacaagtacgccgcctcct cctacctgtccctgacccccgagcagtggaagtc ccaccggtcctactcctgccaggtgacccacgag ggctccaccgtggagaagaccgtggcccccaccg agtgctcctga mAb14315 AminoAcids HCVR QVQLVQSGAEVKKPGSSVKVSCKASGDTFSTYAI 212 NWVRQAPGQGLEWMGRFIHIFGTANYAQKFQGRV TITADESTSTAYMELRSLRSEDTAVYYCARDGVD YGDYRPDYWGQGTLVTVSS HCDR1 GDTFSTYA 214 HCDR2 FIHIFGTA 216 HCDR3 ARDGVDYGDYRPDY 218 LCVR EIVLTQSPGTLSLSPGERATLSCRASQSVSSNYL 220 AWYQQKPGQAPRLLIYGASSRATGIPERFSGSGS GTDFTLTISRLEPEDFAVYYCQQYGSSLYTFGQG TKLEIK LCDR1 QSVSSNY 222 LCDR2 GAS 126 LCDR3 QQYGSSLYT 224 HC QVQLVQSGAEVKKPGSSVKVSCKASGDTFSTYAI 226 NWVRQAPGQGLEWMGRFIHIFGTANYAQKFQGRV TITADESTSTAYMELRSLRSEDTAVYYCARDGVD YGDYRPDYWGQGTLVTVSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSG VHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYIC NVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEL LGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRV VSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSF FLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGK LC EIVLTQSPGTLSLSPGERATLSCRASQSVSSNYL 228 AWYQQKPGQAPRLLIYGASSRATGIPERFSGSGS GTDFTLTISRLEPEDFAVYYCQQYGSSLYTFGQG TKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCL LNNFYPREAKVQWKVDNALQSGNSQESVTEQDSK DSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSS PVTKSFNRGEC NucleicAcids HCVR caggtgcagctggtgcagtctggggctgaggtga 211 agaagcctgggtcctcggtgaaggtctcctgcaa ggcttctggagacaccttcagcacctatgctatc aactgggtgcgacaggcccctggacaagggcttg agtggatgggaagattcatccatatctttggtac agcaaactacgcacagaagttccagggcagagtc accattaccgcggacgaatccacgagcacagcct acatggagctgcgcagcctgagatctgaggacac ggccgtttattactgcgcgagagacggagtagac tacggtgactaccgacctgactactggggccagg gaaccctggtcaccgtctcctca HCDR1 ggagacaccttcagcacctatgct 213 HCDR2 ttcatccatatctttggtacagca 215 HCDR3 gcgagagacggagtagactacggtgactaccgac 217 ctgactac LCVR gaaattgtgttgacgcagtctccaggcaccctgt 219 ctttgtctccaggggaaagagccaccctctcctg cagggccagtcagagtgttagtagcaactactta gcctggtaccagcagaaacctggccaggctccca gactcctcatctatggtgcatccagcagggccac tggcatcccagagaggttcagtggcagtgggtct gggacagacttcactctcaccatcagcagactgg agcctgaagattttgcagtatattactgtcagca gtatggtagctcgctgtacacttttggccagggg accaagctggagatcaaa LCDR1 cagagtgttagtagcaactac 221 LCDR2 ggtgcatcc 125 LCDR3 cagcagtatggtagctcgctgtacact 223 HC caggtgcagctggtgcagtctggggctgaggtga 225 agaagcctgggtcctcggtgaaggtctcctgcaa ggcttctggagacaccttcagcacctatgctatc aactgggtgcgacaggcccctggacaagggcttg agtggatgggaagattcatccatatctttggtac agcaaactacgcacagaagttccagggcagagtc accattaccgcggacgaatccacgagcacagcct acatggagctgcgcagcctgagatctgaggacac ggccgtttattactgcgcgagagacggagtagac tacggtgactaccgacctgactactggggccagg gaaccctggtcaccgtctcctcagcctccaccaa gggcccatcggtcttccccctggcaccctcctcc aagagcacctctgggggcacagcggccctgggct gcctggtcaaggactacttccccgaaccggtgac ggtgtcgtggaactcaggcgccctgaccagcggc gtgcacaccttcccggctgtcctacagtcctcag gactctactccctcagcagcgtggtgaccgtgcc ctccagcagcttgggcacccagacctacatctgc aacgtgaatcacaagcccagcaacaccaaggtgg acaagaaagttgagcccaaatcttgtgacaaaac tcacacatgcccaccgtgcccagcacctgaactc ctggggggaccgtcagtcttcctcttccccccaa aacccaaggacaccctcatgatctcccggacccc tgaggtcacatgcgtggtggtggacgtgagccac gaagaccctgaggtcaagttcaactggtacgtgg acggcgtggaggtgcataatgccaagacaaagcc gcgggaggagcagtacaacagcacgtaccgtgtg gtcagcgtcctcaccgtcctgcaccaggactggc tgaatggcaaggagtacaagtgcaaggtctccaa caaagccctcccagcccccatcgagaaaaccatc tccaaagccaaagggcagccccgagaaccacagg tgtacaccctgcccccatcccgggatgagctgac caagaaccaggtcagcctgacctgcctggtcaaa ggcttctatcccagcgacatcgccgtggagtggg agagcaatgggcagccggagaacaactacaagac cacgcctcccgtgctggactccgacggctccttc ttcctctacagcaagctcaccgtggacaagagca ggtggcagcaggggaacgtcttctcatgctccgt gatgcatgaggctctgcacaaccactacacgcag aagtccctctccctgtctccgggtaaatga LC gaaattgtgttgacgcagtctccaggcaccctgt 227 ctttgtctccaggggaaagagccaccctctcctg cagggccagtcagagtgttagtagcaactactta gcctggtaccagcagaaacctggccaggctccca gactcctcatctatggtgcatccagcagggccac tggcatcccagagaggttcagtggcagtgggtct gggacagacttcactctcaccatcagcagactgg agcctgaagattttgcagtatattactgtcagca gtatggtagctcgctgtacacttttggccagggg accaagctggagatcaaacgaactgtggctgcac catctgtcttcatcttcccgccatctgatgagca gttgaaatctggaactgcctctgttgtgtgcctg ctgaataacttctatcccagagaggccaaagtac agtggaaggtggataacgccctccaatcgggtaa ctcccaggagagtgtcacagagcaggacagcaag gacagcacctacagcctcagcagcaccctgacgc tgagcaaagcagactacgagaaacacaaagtcta cgcctgcgaagtcacccatcagggcctgagctcg cccgtcacaaagagcttcaacaggggagagtgtt ag mAb15151 AminoAcids HCVR QVQLVESGGGVVQPGRSLRLSCAASGFVFNSYGM 270 HWVRQAPGKGLEWVAVLWYEGSKNYYADSVKGRF TISRDNSKNTLYLQMNSLRAEDTAVYYCARHGSG SFFGYYLDYWGQGTLVTVSS HCDR1 GFVFNSYG 272 HCDR2 LWYEGSKN 274 HCDR3 ARHGSGSFFGYYLDY 276 LCVR DIQMTQSPSSLSASVGDRVTITCRASHNINDFLN 278 WYQQKPGKAPRLLIYAASSLQSGVPSRFSGSGSG TDFTLTISSLQPEDFATYYCQESYTTPPTFGQGT KLEIK LCDR1 HNINDF 280 LCDR2 AAS 106 LCDR3 QESYTTPPT 282 HC QVQLVESGGGVVQPGRSLRLSCAASGFVFNSYGM 284 HWVRQAPGKGLEWVAVLWYEGSKNYYADSVKGRF TISRDNSKNTLYLQMNSLRAEDTAVYYCARHGSG SFFGYYLDYWGQGTLVTVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTS GVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYI CNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPE LLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYR VVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYT QKSLSLSPGK LC DIQMTQSPSSLSASVGDRVTITCRASHNINDFLN 286 WYQQKPGKAPRLLIYAASSLQSGVPSRFSGSGSG TDFTLTISSLQPEDFATYYCQESYTTPPTFGQGT KLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC NucleicAcids HCVR caggtgcagctggtggagtctgggggaggcgtgg 269 tccagcctgggaggtccctgagactctcctgtgc agcgtctggtttcgtcttcaatagctatggcatg cactgggtccgccaggctccaggcaaggggctgg agtgggtggcagttctatggtatgaaggaagtaa aaattactatgcagactccgtgaagggccgattc accatctccagagacaattccaagaacacactgt atctgcaaatgaacagcctgagagccgaggacac ggctgtgtattactgtgcgaggcatggttcaggg agtttttttgggtactacttggactactggggcc agggaaccctggtcaccgtctcctca HCDR1 ggtttcgtcttcaatagctatggc 271 HCDR2 ctatggtatgaaggaagtaaaaat 273 HCDR3 gcgaggcatggttcagggagtttttttgggtact 275 acttggactac LCVR gacatccagatgacccagtctccatcctccctgt 277 ctgcatctgtaggagacagagtcaccatcacttg ccgggcaagtcacaacattaatgactttttaaat tggtatcagcagaaaccagggaaagcccctaggc tcctgatctatgctgcatccagtttgcaaagtgg ggtcccatcaaggttcagtggcagtggatctggg acagatttcactctcaccatcagcagtctacaac ctgaagattttgcaacttactactgtcaagagag ttacactacccctccgacttttggccaggggacc aagctggagatcaaa LCDR1 cacaacattaatgacttt 279 LCDR2 gctgcatcc 105 LCDR3 caagagagttacactacccctccgact 281 HC caggtgcagctggtggagtctgggggaggcgtgg 283 tccagcctgggaggtccctgagactctcctgtgc agcgtctggtttcgtcttcaatagctatggcatg cactgggtccgccaggctccaggcaaggggctgg agtgggtggcagttctatggtatgaaggaagtaa aaattactatgcagactccgtgaagggccgattc accatctccagagacaattccaagaacacactgt atctgcaaatgaacagcctgagagccgaggacac ggctgtgtattactgtgcgaggcatggttcaggg agtttttttgggtactacttggactactggggcc agggaaccctggtcaccgtctcctcagcctccac caagggcccatcggtcttccccctggcaccctcc tccaagagcacctctgggggcacagcggccctgg gctgcctggtcaaggactacttccccgaaccggt gacggtgtcgtggaactcaggcgccctgaccagc ggcgtgcacaccttcccggctgtcctacagtcct caggactctactccctcagcagcgtggtgaccgt gccctccagcagcttgggcacccagacctacatc tgcaacgtgaatcacaagcccagcaacaccaagg tggacaagaaagttgagcccaaatcttgtgacaa aactcacacatgcccaccgtgcccagcacctgaa ctcctggggggaccgtcagtcttcctcttccccc caaaacccaaggacaccctcatgatctcccggac ccctgaggtcacatgcgtggtggtggacgtgagc cacgaagaccctgaggtcaagttcaactggtacg tggacggcgtggaggtgcataatgccaagacaaa gccgcgggaggagcagtacaacagcacgtaccgt gtggtcagcgtcctcaccgtcctgcaccaggact ggctgaatggcaaggagtacaagtgcaaggtctc caacaaagccctcccagcccccatcgagaaaacc atctccaaagccaaagggcagccccgagaaccac aggtgtacaccctgcccccatcccgggatgagct gaccaagaaccaggtcagcctgacctgcctggtc aaaggcttctatcccagcgacatcgccgtggagt gggagagcaatgggcagccggagaacaactacaa gaccacgcctcccgtgctggactccgacggctcc ttcttcctctacagcaagctcaccgtggacaaga gcaggtggcagcaggggaacgtcttctcatgctc cgtgatgcatgaggctctgcacaaccactacacg cagaagtccctctccctgtctccgggtaaatga LC gacatccagatgacccagtctccatcctccctgt 285 ctgcatctgtaggagacagagtcaccatcacttg ccgggcaagtcacaacattaatgactttttaaat tggtatcagcagaaaccagggaaagcccctaggc tcctgatctatgctgcatccagtttgcaaagtgg ggtcccatcaaggttcagtggcagtggatctggg acagatttcactctcaccatcagcagtctacaac ctgaagattttgcaacttactactgtcaagagag ttacactacccctccgacttttggccaggggacc aagctggagatcaaacgaactgtggctgcaccat ctgtcttcatcttcccgccatctgatgagcagtt gaaatctggaactgcctctgttgtgtgcctgctg aataacttctatcccagagaggccaaagtacagt ggaaggtggataacgccctccaatcgggtaactc ccaggagagtgtcacagagcaggacagcaaggac agcacctacagcctcagcagcaccctgacgctga gcaaagcagactacgagaaacacaaagtctacgc ctgcgaagtcacccatcagggcctgagctcgccc gtcacaaagagcttcaacaggggagagtgttag mAb14284 AminoAcids HCVR QITLKESGPTLVKPTQTLTLTCTFSGFSFSTSGV 493 GVGWIRQPPGKTLEWLALIYWDDDKRYSPSLKSR LTITKDTSKNQVVLTMTNMDPVDTATYFCAHHGI PT1FGYWGQGALVTVSS HCDR1 GFSFSTSGVG 495 HCDR2 IYWDDDK 497 HCDR3 AHHGIPTIFGY 499 LCVR QSALTQPASVSGSPGQSITISCTGTSSDLGVFNY 501 VSWYQQHPGKAPKLMIYEVTNRPSGVSNRFSGSK SGNTASLTISGLQAEDEADYYCSSYTTSSTVFGG GTKLTVL LCDR1 SSDLGVFNY 503 LCDR2 EVT 505 LCDR3 SSYTTSSTV 507 HC QITLKESGPTLVKPTQTLTLTCTFSGFSFSTSGV 509 GVGWIRQPPGKTLEWLALIYWDDDKRYSPSLKSR LTITKDTSKNQVVLTMTNMDPVDTATYFCAHHGI PTIFGYWGQGALVTVSSASTKGPSVFPLAPSSKS TSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVH TFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLG GPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHED PEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVS VLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISK AKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGF YPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFL YSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKS LSLSPGK LC QSALTQPASVSGSPGQSITISCTGTSSDLGVFNY 511 VSWYQQHPGKAPKLMIYEVTNRPSGVSNRFSGSK SGNTASLTISGLQAEDEADYYCSSYTTSSTVFGG GTKLTVLGQPKAAPSVTLFPPSSEELQANKATLV CLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQ SNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGST VEKTVAPTECS NucleicAcids HCVR cagatcaccttgaaggagtctggtcctacgctgg 492 tgaaacccacacagaccctcacgctgacctgcac cttctctgggttctcatttagcacttctggagtg ggtgtgggctggatccgtcagcccccaggaaaga ccctggagtggcttgcactcatttattgggatga tgataagcgctacagcccatctctgaagagcagg ctcaccattaccaaggacacctccaaaaaccagg tggtccttacaatgaccaacatggaccctgtgga cactgccacatatttctgtgcacaccatggaata cctacgatctttggctactggggccagggagccc tggtcaccgtctcctca HCDR1 gggttctcatttagcacttctggagtgggt 494 HCDR2 atttattgggatgatgataag 496 HCDR3 gcacaccatggaatacctacgatctttggctac 498 LCVR cagtctgccctgactcagcctgcctccgtgtctg 500 ggtctcctggacagtcgatcaccatctcctgcac tggaaccagcagtgaccttggtgtttttaactat gtctcctggtaccaacagcacccaggcaaagccc ccaaactcatgatttatgaggtcactaatcggcc ctcaggggtttctaatcgcttctctggctccaag tctggcaacacggcctccctgaccatctctgggc tccaggctgaggacgaggctgattattattgcag ctcatatacaaccagcagcactgttttcggcgga gggaccaagctgaccgtccta LCDR1 agcagtgaccttggtgtttttaactat 502 LCDR2 gaggtcact 504 LCDR3 agctcatatacaaccagcagcactgtt 506 HC cagatcaccttgaaggagtctggtcctacgctgg 508 tgaaacccacacagaccctcacgctgacctgcac cttctctgggttctcatttagcacttctggagtg ggtgtgggctggatccgtcagcccccaggaaaga ccctggagtggcttgcactcatttattgggatga tgataagcgctacagcccatctctgaagagcagg ctcaccattaccaaggacacctccaaaaaccagg tggtccttacaatgaccaacatggaccctgtgga cactgccacatatttctgtgcacaccatggaata cctacgatctttggctactggggccagggagccc tggtcaccgtctcctcagcctccaccaagggccc atcggtcttccccctggcaccctcctccaagagc acctctgggggcacagcggccctgggctgcctgg tcaaggactacttccccgaaccggtgacggtgtc gtggaactcaggcgccctgaccagcggcgtgcac accttcccggctgtcctacagtcctcaggactct actccctcagcagcgtggtgaccgtgccctccag cagcttgggcacccagacctacatctgcaacgtg aatcacaagcccagcaacaccaaggtggacaaga aagttgagcccaaatcttgtgacaaaactcacac atgcccaccgtgcccagcacctgaactcctgggg ggaccgtcagtcttcctcttccccccaaaaccca aggacaccctcatgatctcccggacccctgaggt cacatgcgtggtggtggacgtgagccacgaagac cctgaggtcaagttcaactggtacgtggacggcg tggaggtgcataatgccaagacaaagccgcggga ggagcagtacaacagcacgtaccgtgtggtcagc gtcctcaccgtcctgcaccaggactggctgaatg gcaaggagtacaagtgcaaggtctccaacaaagc cctcccagcccccatcgagaaaaccatctccaaa gccaaagggcagccccgagaaccacaggtgtaca ccctgcccccatcccgggatgagctgaccaagaa ccaggtcagcctgacctgcctggtcaaaggcttc tatcccagcgacatcgccgtggagtgggagagca atgggcagccggagaacaactacaagaccacgcc tcccgtgctggactccgacggctccttcttcctc tacagcaagctcaccgtggacaagagcaggtggc agcaggggaacgtcttctcatgctccgtgatgca tgaggctctgcacaaccactacacgcagaagtcc ctctccctgtctccgggtaaatga LC cagtctgccctgactcagcctgcctccgtgtctg 510 ggtctcctggacagtcgatcaccatctcctgcac tggaaccagcagtgaccttggtgtttttaactat gtctcctggtaccaacagcacccaggcaaagccc ccaaactcatgatttatgaggtcactaatcggcc ctcaggggtttctaatcgcttctctggctccaag tctggcaacacggcctccctgaccatctctgggc tccaggctgaggacgaggctgattattattgcag ctcatatacaaccagcagcactgttttcggcgga gggaccaagctgaccgtcctaggccagcccaagg ccgccccctccgtgaccctgttccccccctcctc cgaggagctgcaggccaacaaggccaccctggtg tgcctgatctccgacttctaccccggcgccgtga ccgtggcctggaaggccgactcctcccccgtgaa ggccggcgtggagaccaccaccccctccaagcag tccaacaacaagtacgccgcctcctcctacctgt ccctgacccccgagcagtggaagtcccaccggtc ctactcctgccaggtgacccacgagggctccacc gtggagaagaccgtggcccccaccgagtgctcct ga mAb15160 AminoAcids HCVR EVQLVESGGGLVQPGGSLRLSCSASGFTFSRYAM 362 YWVRQAPGKGLEYVSAISSDGGSTYDADSVKGRF TISRANSKNTLYLQMSSLRAEDTAVYYCVKGLRE LLYYYYGMDVWGQGTTVTVSS HCDR1 GFTFSRYA 364 HCDR2 ISSDGGST 366 HCDR3 VKGLRELLYYYYGMDV 368 LCVR DIQMTQSPSSLSASVGDRVTITCRAGQSISSFLN 370 WYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSG TDFTLTISSLQPEDFATYYCQQSYITPFTFGPGT KVDIK LCDR1 QSISSF 372 LCDR2 AAS 106 LCDR3 QQSYITPFT 374 HC EVQLVESGGGLVQPGGSLRLSCSASGFTFSRYAM 376 YWVRQAPGKGLEYVSAISSDGGSTYDADSVKGRF TISRANSKNTLYLQMSSLRAEDTAVYYCVKGLRE LLYYYYGMDVWGQGTTVTVSSASTKGPSVFPLAP SSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALT SGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTY ICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCL VKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDG SFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK LC DIQMTQSPSSLSASVGDRVTITCRAGQSISSFLN 378 WYQQKPGKAPKLLIYAASSLQSGVPSRFSGSGSG TDFTLTISSLQPEDFATYYCQQSYITPFTFGPGT KVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC NucleicAcids HCVR gaggtgcagctggtggagtctgggggaggcttgg 361 tccagcctggggggtccctgagactctcctgttc agcctctggattcaccttcagtaggtacgctatg tactgggtccgccaggctccagggaagggactgg aatatgtttcagctattagtagtgatgggggtag cacatacgacgcagactccgtgaagggcagattc accatctccagagccaattccaagaacacgctgt accttcaaatgagcagtctgagagctgaggacac ggctgtgtattattgtgtgaaaggtctgcgggag ttactctactactattacggaatggacgtctggg gccaagggactacggtcaccgtctcctca HCDR1 ggattcaccttcagtaggtacgct 363 HCDR2 attagtagtgatgggggtagcaca 365 HCDR3 gtgaaaggtctgcgggagttactctactactatt 367 acggaatggacgtc LCVR gacatccagatgacccagtctccatcctccctgt 369 ctgcatctgtaggagacagagtcaccatcacttg ccgggcaggtcagagcattagcagctttttaaat tggtatcagcagaagccagggaaagcccctaagc tcctgatctatgctgcatccagtttgcaaagtgg ggtcccatcaaggttcagtggcagtggatctggg acagatttcactctcaccatcagcagtctccaac ctgaagattttgcaacttactactgtcaacagag ttacattacccccttcactttcggccctgggacc aaggtggatatcaaa LCDR1 cagagcattagcagcttt 371 LCDR2 gctgcatcc 105 LCDR3 caacagagttacattacccccttcact 373 HC gaggtgcagctggtggagtctgggggaggcttgg 375 tccagcctggggggtccctgagactctcctgttc agcctctggattcaccttcagtaggtacgctatg tactgggtccgccaggctccagggaagggactgg aatatgtttcagctattagtagtgatgggggtag cacatacgacgcagactccgtgaagggcagattc accatctccagagccaattccaagaacacgctgt accttcaaatgagcagtctgagagctgaggacac ggctgtgtattattgtgtgaaaggtctgcgggag ttactctactactattacggaatggacgtctggg gccaagggactacggtcaccgtctcctcagcctc caccaagggcccatcggtcttccccctggcaccc tcctccaagagcacctctgggggcacagcggccc tgggctgcctggtcaaggactacttccccgaacc ggtgacggtgtcgtggaactcaggcgccctgacc agcggcgtgcacaccttcccggctgtcctacagt cctcaggactctactccctcagcagcgtggtgac cgtgccctccagcagcttgggcacccagacctac atctgcaacgtgaatcacaagcccagcaacacca aggtggacaagaaagttgagcccaaatcttgtga caaaactcacacatgcccaccgtgcccagcacct gaactcctggggggaccgtcagtcttcctcttcc ccccaaaacccaaggacaccctcatgatctcccg gacccctgaggtcacatgcgtggtggtggacgtg agccacgaagaccctgaggtcaagttcaactggt acgtggacggcgtggaggtgcataatgccaagac aaagccgcgggaggagcagtacaacagcacgtac cgtgtggtcagcgtcctcaccgtcctgcaccagg actggctgaatggcaaggagtacaagtgcaaggt ctccaacaaagccctcccagcccccatcgagaaa accatctccaaagccaaagggcagccccgagaac cacaggtgtacaccctgcccccatcccgggatga gctgaccaagaaccaggtcagcctgacctgcctg gtcaaaggcttctatcccagcgacatcgccgtgg agtgggagagcaatgggcagccggagaacaacta caagaccacgcctcccgtgctggactccgacggc tccttcttcctctacagcaagctcaccgtggaca agagcaggtggcagcaggggaacgtcttctcatg ctccgtgatgcatgaggctctgcacaaccactac acgcagaagtccctctccctgtctccgggtaaat ga LC gacatccagatgacccagtctccatcctccctgt 377 ctgcatctgtaggagacagagtcaccatcacttg ccgggcaggtcagagcattagcagctttttaaat tggtatcagcagaagccagggaaagcccctaagc tcctgatctatgctgcatccagtttgcaaagtgg ggtcccatcaaggttcagtggcagtggatctggg acagatttcactctcaccatcagcagtctccaac ctgaagattttgcaacttactactgtcaacagag ttacattacccccttcactttcggccctgggacc aaggtggatatcaaacgaactgtggctgcaccat ctgtcttcatcttcccgccatctgatgagcagtt gaaatctggaactgcctctgttgtgtgcctgctg aataacttctatcccagagaggccaaagtacagt ggaaggtggataacgccctccaatcgggtaactc ccaggagagtgtcacagagcaggacagcaaggac agcacctacagcctcagcagcaccctgacgctga gcaaagcagactacgagaaacacaaagtctacgc ctgcgaagtcacccatcagggcctgagctcgccc gtcacaaagagcttcaacaggggagagtgttag

Administration of Antibodies

[0150] The present invention provides methods for administering an anti-CoV-S antigen-binding protein of the present invention, e.g., those of Table 4, comprising introducing the antigen-binding protein into the body of a subject (e.g., a human). For example, the method comprises piercing the body of the subject with a needle of a syringe and injecting the antigen-binding protein into the body of the subject, e.g., into the vein, artery, tumor, muscular tissue or subcutis of the subject.

[0151] The present invention provides a vessel (e.g., a plastic or glass vial, e.g., with a cap or a chromatography column, hollow bore needle or a syringe cylinder) comprising an anti-CoV-S antigen-binding protein of the present invention, e.g., those of Table 4.

[0152] The present invention also provides an injection device comprising one or more antigen-binding proteins (e.g., antibody or antigen-binding fragment) that bind specifically to CoV-S, e.g., those of Table 4, or a pharmaceutical composition thereof. The injection device may be packaged into a kit. An injection device is a device that introduces a substance into the body of a subject via a parenteral route, e.g., intramuscular, subcutaneous or intravenous. For example, an injection device may be a syringe (e.g., pre-filled with the pharmaceutical composition, such as an auto-injector) which, for example, includes a cylinder or barrel for holding fluid to be injected (e.g., comprising the antibody or fragment or a pharmaceutical composition thereof), a needle for piecing skin and/or blood vessels for injection of the fluid; and a plunger for pushing the fluid out of the cylinder and through the needle bore. In an embodiment of the invention, an injection device that comprises an antigen-binding protein, e.g., an antibody or antigen-binding fragment thereof, from a combination of the present invention, or a pharmaceutical composition thereof is an intravenous (IV) injection device. Such a device can include the antigen-binding protein or a pharmaceutical composition thereof in a cannula or trocar/needle which may be attached to a tube which may be attached to a bag or reservoir for holding fluid (e.g., saline) introduced into the body of the subject through the cannula or trocar/needle. The antibody or fragment or a pharmaceutical composition thereof may, in an embodiment of the invention, be introduced into the device once the trocar and cannula are inserted into the vein of a subject and the trocar is removed from the inserted cannula. The IV device may, for example, be inserted into a peripheral vein (e.g., in the hand or arm); the superior vena cava or inferior vena cava, or within the right atrium of the heart (e.g., a central IV); or into a subclavian, internal jugular, or a femoral vein and, for example, advanced toward the heart until it reaches the superior vena cava or right atrium (e.g., a central venous line). In an embodiment of the invention, an injection device is an autoinjector, a jet injector or an external infusion pump. A jet injector uses a high-pressure narrow jet of liquid which penetrate the epidermis to introduce the antibody or fragment or a pharmaceutical composition thereof to a subject's body. External infusion pumps are medical devices that deliver the antibody or fragment or a pharmaceutical composition thereof into a subject's body in controlled amounts. External infusion pumps may be powered electrically or mechanically. Different pumps operate in different ways, for example, a syringe pump holds fluid in the reservoir of a syringe, and a moveable piston controls fluid delivery, an elastomeric pump holds fluid in a stretchable balloon reservoir, and pressure from the elastic walls of the balloon drives fluid delivery. In a peristaltic pump, a set of rollers pinches down on a length of flexible tubing, pushing fluid forward. In a multi-channel pump, fluids can be delivered from multiple reservoirs at multiple rates.

Preparation of Human Antibodies

[0153] Methods for generating human antibodies in transgenic mice are known in the art. Any such known methods can be used in the context of the present invention to make human antibodies that specifically bind to CoV-S. An immunogen comprising any one of the following can be used to generate antibodies to CoV-S. In certain embodiments of the invention, the antibodies of the invention are obtained from mice immunized with a full length, native CoV-S, or with a live attenuated or inactivated virus, or with DNA encoding the protein or fragment thereof. Alternatively, the CoV-S protein or a fragment thereof may be produced using standard biochemical techniques and modified and used as immunogen. In one embodiment of the invention, the immunogen is a recombinantly produced CoV-S protein or fragment thereof. In certain embodiments of the invention, the immunogen may be a CoV-S polypeptide vaccine. In certain embodiments, one or more booster injections may be administered. In certain embodiments, the immunogen may be a recombinant CoV-S polypeptide expressed in E. coli or in any other eukaryotic or mammalian cells such as Chinese hamster ovary (CHO) cells.

[0154] Using VELOCIMMUNE technology (see, for example, U.S. Pat. No. 6,596,541, Regeneron Pharmaceuticals, VELOCIMMUNE) or any other known method for generating monoclonal antibodies, high affinity chimeric antibodies to CoV-S can be initially isolated having a human variable region and a mouse constant region. The VELOCIMMUNE technology involves generation of a transgenic mouse having a genome comprising human heavy and light chain variable regions operably linked to endogenous mouse constant region loci such that the mouse produces an antibody comprising a human variable region and a mouse constant region in response to antigenic stimulation. The DNA encoding the variable regions of the heavy and light chains of the antibody are isolated and operably linked to DNA encoding the human heavy and light chain constant regions. The DNA is then expressed in a cell capable of expressing the fully human antibody.

[0155] Generally, a VELOCIMMUNE mouse is challenged with the antigen of interest, and lymphatic cells (such as B-cells) are recovered from the mice that express antibodies. The lymphatic cells may be fused with a myeloma cell line to prepare immortal hybridoma cell lines, and such hybridoma cell lines are screened and selected to identify hybridoma cell lines that produce antibodies specific to the antigen of interest. DNA encoding the variable regions of the heavy chain and light chain may be isolated and linked to desirable isotypic constant regions of the heavy chain and light chain. Such an antibody protein may be produced in a cell, such as a CHO cell. Alternatively, DNA encoding the antigen-specific chimeric antibodies or the variable domains of the light and heavy chains may be isolated directly from antigen-specific lymphocytes.

[0156] Initially, high affinity chimeric antibodies are isolated having a human variable region and a mouse constant region. As in the experimental section below, the antibodies are characterized and selected for desirable characteristics, including affinity, selectivity, epitope, etc. The mouse constant regions are replaced with a desired human constant region to generate the fully human antibody of the invention, for example wild-type or modified IgG1 or IgG4. While the constant region selected may vary according to specific use, high affinity antigen-binding and target specificity characteristics reside in the variable region.

Anti-Coronavirus Spike Protein Antibodies Comprising Fc Variants

[0157] According to certain embodiments of the present invention, anti-CoV-S antigen-binding proteins, e.g., antibodies or antigen-binding fragments, are provided comprising an Fc domain comprising one or more mutations, which, for example, enhance or diminish antibody binding to the FcRn receptor, e.g., at acidic pH as compared to neutral pH. For example, the present invention includes anti-CoV-S antibodies comprising a mutation in the C.sub.H2 or a C.sub.H3 region of the Fc domain, wherein the mutation(s) increases the affinity of the Fc domain to FcRn in an acidic environment (e.g., in an endosome where pH ranges from about 5.5 to about 6.0). Such mutations may result in an increase in serum half-life of the antibody when administered to an animal. Non-limiting examples of such Fc modifications include, e.g., a modification at position 250 (e.g., E or Q); 250 and 428 (e.g., L or F); 252 (e.g., UY/F/W or T), 254 (e.g., S or T), and 256 (e.g., S/R/Q/E/D or T); or a modification at position 428 and/or 433 (e.g., H/L/R/S/P/Q or K) and/or 434 (e.g., A, W, H, F or Y [N434A, N434W, N434H, N434F or N434Y]); or a modification at position 250 and/or 428; or a modification at position 307 or 308 (e.g., 308F, V308F), and 434. In one embodiment, the modification comprises a 428L (e.g., M428L) and 434S (e.g., N434S) modification; a 428L, 259I (e.g., V259I), and 308F (e.g., V308F) modification; a 433K (e.g., H433K) and a 434 (e.g., 434Y) modification; a 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modification; a 250Q and 428L modification (e.g., T250Q and M428L); and a 307 and/or 308 modification (e.g., 308F or 308P). In yet another embodiment, the modification comprises a 265A (e.g., D265A) and/or a 297A (e.g., N297A) modification.

[0158] For example, the present invention includes anti-CoV-S antigen-binding proteins, e.g., antibodies or antigen-binding fragments, comprising an Fc domain comprising one or more pairs or groups of mutations selected from the group consisting of: 250Q and 248L (e.g., T250Q and M248L); 252Y, 254T and 256E (e.g., M252Y, S254T and T256E); 428L and 434S (e.g., M428L and N434S); 257I and 311I (e.g., P257I and Q311I); 257I and 434H (e.g., P257I and N434H); 376V and 434H (e.g., D376V and N434H); 307A, 380A and 434A (e.g., T307A, E380A and N434A); and 433K and 434F (e.g., H433K and N434F). In particular, antibodies designated mAb17090 and mAb15160_2 each contain M252Y, S254T and T256E modifications in the heavy chain constant region as compared to mAb14286 and mAb15160, respectively.

[0159] Anti-CoV-S antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof, that comprise a V.sub.H and/or V.sub.L as set forth herein comprising any possible combinations of the foregoing Fc domain mutations, are contemplated within the scope of the present invention.

[0160] The present invention also includes anti-CoV-S antigen-binding proteins, antibodies or antigen-binding fragments, comprising a V.sub.H set forth herein and a chimeric heavy chain constant (C.sub.H) region, wherein the chimeric C.sub.H region comprises segments derived from the C.sub.H regions of more than one immunoglobulin isotype. For example, the antibodies of the invention may comprise a chimeric C.sub.H region comprising part or all of a C.sub.H.sup.2 domain derived from a human IgG1, human IgG2 or human IgG4 molecule, combined with part or all of a C.sub.H3 domain derived from a human IgG1, human IgG2 or human IgG4 molecule. According to certain embodiments, the antibodies of the invention comprise a chimeric C.sub.H region having a chimeric hinge region. For example, a chimeric hinge may comprise an upper hinge amino acid sequence (amino acid residues from positions 216 to 227 according to EU numbering) derived from a human IgG1, a human IgG2 or a human IgG4 hinge region, combined with a lower hinge sequence (amino acid residues from positions 228 to 236 according to EU numbering) derived from a human IgG1, a human IgG2 or a human IgG4 hinge region. According to certain embodiments, the chimeric hinge region comprises amino acid residues derived from a human IgG1 or a human IgG4 upper hinge and amino acid residues derived from a human IgG2 lower hinge. An antibody comprising a chimeric C.sub.H region as described herein may, in certain embodiments, exhibit modified Fc effector functions without adversely affecting the therapeutic or pharmacokinetic properties of the antibody. (See, e.g., WO2014/022540).

Immunoconjugates

[0161] The invention encompasses an anti-CoV-S antigen-binding proteins, e.g., antibodies or antigen-binding fragments, conjugated to another moiety, e.g., a therapeutic moiety (an immunoconjugate), such as a toxoid or an anti-viral drug to treat influenza virus infection. In an embodiment of the invention, an anti-CoV-S antibody or fragment is conjugated to any of the further therapeutic agents set forth herein. As used herein, the term immunoconjugate refers to an antigen-binding protein, e.g., an antibody or antigen-binding fragment, which is chemically or biologically linked to a radioactive agent, a cytokine, an interferon, a target or reporter moiety, an enzyme, a peptide or protein or a therapeutic agent. The antigen-binding protein may be linked to the radioactive agent, cytokine, interferon, target or reporter moiety, enzyme, peptide or therapeutic agent at any location along the molecule so long as it is able to bind its target (CoV-S). Examples of immunoconjugates include antibody-drug conjugates and antibody-toxin fusion proteins. In one embodiment of the invention, the agent may be a second, different antibody that binds specifically to CoV-S. The type of therapeutic moiety that may be conjugated to the anti-CoV-S antigen-binding protein (e.g., antibody or fragment) will take into account the condition to be treated and the desired therapeutic effect to be achieved. See, e.g., Arnon et al., Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy, Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., Antibodies For Drug Delivery, Controlled Drug Delivery (2.sup.nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review, Monoclonal Antibodies 1984: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy, Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates, Immunol. Rev., 62: 119-58 (1982).

Multi-Specific Antibodies

[0162] The present invention includes anti-CoV-S antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof, as well as methods of use thereof and methods of making such antigen-binding proteins. The term anti-CoV-S antigen-binding proteins, e.g., antibodies or antigen-binding fragments, includes multispecific (e.g., bispecific or biparatopic) molecules that include at least one first antigen-binding domain that specifically binds to CoV-S (e.g., an antigen-binding domain from an antibody of Table 4) and at least one second antigen-binding domain that binds to a different antigen or to an epitope in CoV-S which is different from that of the first antigen-binding domain. In some embodiments, the first antigen-binding domain and the second antigen-binding domain are both selected from the antigen-binding domains of Table 4. In an embodiment of the invention, the first and second epitopes overlap. In another embodiment of the invention, the first and second epitopes do not overlap. For example, in an embodiment of the invention, a multispecific antibody is a bispecific IgG antibody (e.g., IgG1 or IgG4) that includes a first antigen-binding domain that binds specifically to CoV-S including the heavy and light immunoglobulin chain of an antibody of Table 4, and a second antigen-binding domain that binds specifically to a different epitope of CoV-S. In some embodiments, a bispecific IgG antibody (e.g., IgG1 or IgG4) includes a first antigen-binding domain that binds specifically to CoV-S and a second binding domain that binds to a host cell protein, e.g., ACE2 or TMPRSS2.

[0163] The antibodies of Table 4 include multispecific molecules, e.g., antibodies or antigen-binding fragments, that include the CDR-Hs and CDR-Ls, V.sub.H and V.sub.L, or HC and LC of those antibodies, respectively (including variants thereof as set forth herein).

[0164] In an embodiment of the invention, an antigen-binding domain that binds specifically to CoV-S, which may be included in a multispecific molecule, comprises: [0165] (1) [0166] (i) a heavy chain variable domain sequence that comprises HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in Table 4, and [0167] (ii) a light chain variable domain sequence that comprises LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in Table 4; [0168] or, [0169] (2) [0170] (i) a heavy chain variable domain sequence comprising an amino acid sequence set forth in Table 4, and [0171] (ii) a light chain variable domain sequence comprising an amino acid sequence set forth in Table 4; [0172] or, [0173] (3) [0174] (i) a heavy chain immunoglobulin sequence comprising an amino acid sequence set forth in Table 4, and [0175] (ii) a light chain immunoglobulin sequence comprising an amino acid sequence set forth in Table 4.

[0176] In an embodiment of the invention, the multispecific antibody or fragment includes more than two different binding specificities (e.g., a trispecific molecule), for example, one or more additional antigen-binding domains which are the same or different from the first and/or second antigen-binding domain.

[0177] In one embodiment of the invention, a bispecific antigen-binding fragment comprises a first scFv (e.g., comprising V.sub.H and V.sub.L sequences of Table 4) having binding specificity for a first epitope (e.g., CoV-S) and a second scFv having binding specificity for a second, different epitope. For example, in an embodiment of the invention, the first and second scFv are tethered with a linker, e.g., a peptide linker (e.g., a GS linker such as (GGGGS).sub.n (SEQ ID NO: 834) wherein n is, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10). Other bispecific antigen-binding fragments include an F(ab).sub.2 of a bispecific IgG antibody which comprises the heavy and light chain CDRs of Table 4 and of another antibody that binds to a different epitope.

Therapeutic Methods

[0178] The present invention provides methods for treating or preventing viral infection (e.g., coronavirus infection) by administering a therapeutically effective amount of anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment, (e.g., of Table 4) to a subject (e.g., a human) in need of such treatment or prevention.

[0179] Coronavirus infection may be treated or prevented, in a subject, by administering an anti-CoV-S antigen-binding protein of the present invention to a subject. Exemplary coronaviruses include SARS-CoV-2, which may further include variants such as alpha, beta, gamma, delta, and omicron.

[0180] An effective or therapeutically effective dose of anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment (e.g., of Table 4), for treating or preventing a viral infection refers to the amount of the antibody or fragment sufficient to alleviate one or more signs and/or symptoms of the infection in the treated subject, whether by inducing the regression or elimination of such signs and/or symptoms or by inhibiting the progression of such signs and/or symptoms. The dose amount may vary depending upon the age and the size of a subject to be administered, target disease, conditions, route of administration, and the like. In an embodiment of the invention, an effective or therapeutically effective dose of antibody or antigen-binding fragment thereof of the present invention, for treating or preventing viral infection, e.g., in an adult human subject, is about 0.01 to about 200 mg/kg, e.g., up to about 150 mg/kg. In an embodiment of the invention, the dosage is up to about 10.8 or 11 grams (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11 grams). Depending on the severity of the infection, the frequency and the duration of the treatment can be adjusted. In certain embodiments, the antigen-binding protein of the present invention can be administered at an initial dose, followed by one or more secondary doses. In certain embodiments, the initial dose may be followed by administration of a second or a plurality of subsequent doses of antibody or antigen-binding fragment thereof in an amount that can be approximately the same or less than that of the initial dose, wherein the subsequent doses are separated by at least 1 day to 3 days; at least one week, at least 2 weeks; at least 3 weeks; at least 4 weeks; at least 5 weeks; at least 6 weeks; at least 7 weeks; at least 8 weeks; at least 9 weeks; at least 10 weeks; at least 12 weeks; or at least 14 weeks.

[0181] In some embodiments, mAb10933 and mAb10987 (casirivimab and imdevimab, respectively) can be administered to a subject in a method of prophylaxis, e.g., pre-exposure prophylaxis, against symptomatic COVID-19. In some embodiments, the subject is immunocompromised, e.g., has an immunodeficiency such as a primary or secondary immunodeficiency. In some embodiments, the subject has not mounted an effective response to COVID-19 vaccination. Exemplary criteria associated with immune compromise include: active or recent treatment for solid tumor and hematologic malignancies; receipt of solid-organ or recent hematopoietic stem cell transplants; severe primary immunodeficiency; advanced or untreated HIV infection; active treatment with high-dose corticosteroids, alkylating agents, antimetabolites, tumor-necrosis (TNF) blockers, and other biologic agents that are immunosuppressive or immunomodulatory; and chronic medical conditions such as asplenia and chronic renal disease, in which some patients with these conditions may be associated with varying degrees of immune deficit. In some embodiments, a subject meets 1 of the following criteria: [0182] a. Solid organ transplant (SOT) or hematopoietic blood cell transplant (HSCT) recipients receiving any immunosuppressive medication [0183] b. Active hematologic malignancies or that have completed therapy within 3 months [0184] c. Solid organ malignancies receiving active treatment with T cell or B cell immunosuppressive therapy [0185] d. Moderate or severe primary immunodeficiency (such as hypogammaglobulinemia, Common variable immune deficiency, severe combined immunodeficiency) [0186] e. HIV and CD4 <200 cells/microliter [0187] f. Patients with rheumatologic disease, autoimmune disease, or multiple sclerosis receiving immunosuppressive therapy that modulates Th1, Th17 or B cell responses [0188] g. Receiving any of the following immunosuppressant drugs for more than 3 weeks: [0189] T cell suppressing agents (eg, 5 mg prednisone equivalent/day during >3 weeks), proteasome inhibitors (such as bortezomib, lenalidomide), alemtuzumab, anti-thymocyte globulin, CAR-T therapy, calcineurin inhibitors) [0190] Alkylating agents [0191] Purine analogs, such as fludarabine or cladribine [0192] B cell depleting agents, such as rituximab and ocrelizumab [0193] mTOR inhibitors [0194] Antimetabolites, such as mycophenolate [0195] JAK inhibitors.

[0196] Exemplary dosing regimens include: [0197] Co-administration of mAb10933 and mAb10987 combination therapy, 1200 mg (600 mg per mAb) on day 1, then 600 mg (300 mg per mAb) subcutaneous (SC) every four weeks (Q4W); [0198] Co-administration of mAb10933 and mAb10987 combination therapy, 300 mg (150 mg per mAb) SC Q4W; and [0199] Co-administration of mAb10933 and mAb10987 combination therapy, 300 mg (150 mg per mAb) SC Q12W.

[0200] As used herein, the term subject refers to a mammal (e.g., rat, mouse, cat, dog, cow, pig, sheep, horse, goat, rabbit), preferably a human, for example, in need of prevention and/or treatment of a disease or disorder such as viral infection or cancer. The subject may have a viral infection, e.g., an influenza infection, or be predisposed to developing an infection. Subjects predisposed to developing an infection, or subjects who may be at elevated risk for contracting an infection (e.g., of coronavirus or influenza virus), include subjects with compromised immune systems because of autoimmune disease, subjects receiving immunosuppressive therapy (for example, following organ transplant), subjects afflicted with human immunodeficiency syndrome (HIV) or acquired immune deficiency syndrome (AIDS), subjects with forms of anemia that deplete or destroy white blood cells, subjects receiving radiation or chemotherapy, or subjects afflicted with an inflammatory disorder. Additionally, subjects of very young (e.g., 5 years of age or younger) or old age (e.g., 65 years of age or older) are at increased risk. Moreover, a subject may be at risk of contracting a viral infection due to proximity to an outbreak of the disease, e.g. subject resides in a densely-populated city or in close proximity to subjects having confirmed or suspected infections of a virus, or choice of employment, e.g. hospital worker, pharmaceutical researcher, traveler to infected area, or frequent flier.

[0201] Treat or treating means to administer an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment of the present invention (e.g., of Table 4), to a subject having one or more signs or symptoms of a disease or infection, e.g., viral infection, for which the antigen-binding protein is effective when administered to the subject at an effective or therapeutically effective amount or dose (as discussed herein).

[0202] The present invention also encompasses prophylactically administering an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment thereof of the present invention (e.g., of Table 4), to a subject who is at risk of viral infection so as to prevent such infection. Passive antibody-based immunoprophylaxis has proven an effective strategy for preventing subject from viral infection. See e.g., Berry et al., Passive broad-spectrum influenza immunoprophylaxis. Influenza Res Treat. 2014; 2014:267594. Epub 2014 Sep. 22; and Jianqiang et al., Passive immune neutralization strategies for prevention and control of influenza A infections, Immunotherapy. 2012 February; 4(2): 175-186; Prabhu et al., Antivir Ther. 2009; 14(7):911-21, Prophylactic and therapeutic efficacy of a chimeric monoclonal antibody specific for H5 hemagglutinin against lethal H5N1 influenza. Prevent or preventing means to administer an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment of the present invention (e.g., of Table 4), to a subject to inhibit the manifestation of a disease or infection (e.g., viral infection) in the body of a subject, for which the antigen-binding protein is effective when administered to the subject at an effective or therapeutically effective amount or dose (as discussed herein). As used herein, prophylaxis can be pre-exposure prophylaxis (e.g., administration of an antibody or antigen-binding fragment described herein to an individual prior to exposure to the SARS-CoV-2 virus), or post-exposure prophylaxis (e.g., administration of an antibody or antigen-binding fragment described herein to an individual prior to exposure to the SARS-CoV-2 virus). In some embodiments, post-exposure prophylaxis can prevent one or more symptoms of COVID-19 despite infection with SARS-CoV-2.

[0203] In an embodiment of the invention, a sign or symptom of a viral infection in a subject is survival or proliferation of virus in the body of the subject, e.g., as determined by viral titer assay (e.g., coronavirus propagation in embryonated chicken eggs or coronavirus spike protein assay). Other signs and symptoms of viral infection are discussed herein.

[0204] As noted above, in some embodiments the subject may be a non-human animal, and the antigen-binding proteins (e.g., antibodies and antigen-binding fragments) discussed herein may be used in a veterinary context to treat and/or prevent disease in the non-human animals (e.g., cats, dogs, pigs, cows, horses, goats, rabbits, sheep, and the like).

[0205] The present invention provides a method for treating or preventing viral infection (e.g., coronavirus infection) or for inducing the regression or elimination or inhibiting the progression of at least one sign or symptom of viral infection such as: [0206] fever or feeling feverish/chills; [0207] cough; [0208] sore throat; [0209] runny or stuffy nose; [0210] sneezing; [0211] muscle or body aches; [0212] headaches; [0213] fatigue (tiredness); [0214] vomiting; [0215] diarrhea; [0216] respiratory tract infection; [0217] chest discomfort; [0218] shortness of breath; [0219] bronchitis; and/or [0220] pneumonia,
which sign or symptom is secondary to viral infection, in a subject in need thereof (e.g., a human), by administering a therapeutically effective amount of anti-CoV-S antigen-binding protein (e.g., of Table 4) to the subject, for example, by injection of the protein into the body of the subject.

Combinations and Pharmaceutical Compositions

[0221] To prepare pharmaceutical compositions of the anti-CoV-S antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof (e.g., of Table 4), antigen-binding protein is admixed with a pharmaceutically acceptable carrier or excipient. See, e.g., Remington's Pharmaceutical Sciences and U.S. Pharmacopeia: National Formulary, Mack Publishing Company, Easton, Pa. (1984); Hardman, et al. (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, N.Y.; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, N.Y.; Avis, et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems, Marcel Dekker, NY; Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, N.Y. In an embodiment of the invention, the pharmaceutical composition is sterile. Such compositions are part of the present invention.

[0222] The scope of the present invention includes desiccated, e.g., freeze-dried, compositions comprising an anti-CoV-S antigen-binding proteins, e.g., antibody or antigen-binding fragment thereof (e.g., of Table 4), or a pharmaceutical composition thereof that includes a pharmaceutically acceptable carrier but substantially lacks water.

[0223] In a further embodiment of the invention, a further therapeutic agent that is administered to a subject in association with an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment thereof (e.g., of Table 4), disclosed herein is administered to the subject in accordance with the Physicians' Desk Reference 2003 (Thomson Healthcare; 57.sup.th edition (Nov. 1, 2002)).

[0224] The mode of administration can vary. Routes of administration include oral, rectal, transmucosal, intestinal, parenteral; intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, insufflation, topical, cutaneous, transdermal or intra-arterial.

[0225] The present invention provides methods for administering an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment thereof (e.g., of Table 4), comprising introducing the protein into the body of a subject. For example, the method comprises piercing the body of the subject with a needle of a syringe and injecting the antigen-binding protein into the body of the subject, e.g., into the vein, artery, tumor, muscular tissue or subcutis of the subject.

[0226] The present invention provides a vessel (e.g., a plastic or glass vial, e.g., with a cap or a chromatography column, hollow bore needle or a syringe cylinder) comprising any of the anti-CoV-S antigen-binding proteins, e.g., antibodies or antigen-binding fragments thereof (e.g., of Table 4), polypeptides (e.g., an HC, LC, V.sub.H or V.sub.L of Table 4) or polynucleotides (e.g., of Table 5) or vectors set forth herein or a pharmaceutical composition thereof comprising a pharmaceutically acceptable carrier.

[0227] In an embodiment of the present disclosure, an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment thereof of the present invention (e.g., of Table 4), is administered in association with one or more further therapeutic agents. A further therapeutic agent includes, but is not limited to: an anti-inflammatory agent, an antimalarial agent, a second antibody or antigen-binding fragment thereof that specifically binds TMPRSS2, and a second antibody or antigen-binding fragment thereof that specifically binds to CoV-S (e.g., an antibody described herein, or described in U.S. Pat. No. 10,787,501 which is hereby specifically incorporated by reference in its entirety). In some embodiments, an antimalarial agent is chloroquine or hydroxychloroquine. In some embodiments, an anti-inflammatory agent is an antibody such as sarilumab, tocilizumab, or gimsilumab. In some embodiments, the further therapeutic agent is a second antibody or antigen-binding fragment disclosed herein, e.g., of Table 4. In some cases, the antibody that binds to CoV-S is casirivimab or imdevimab, In certain embodiments, one, two, three, four, or more antibodies, or antigen-binding fragments thereof, of Table 4 can be administered in combination (e.g., concurrently or sequentially). In particular, combinations of antibodies can be selected such that the antibodies do not cross-compete, as described in Example 7. In some embodiments, mAb14256 is administered in combination with mAb14315. In some embodiments, mAb15151 is administered in combination with mAb14315. In some embodiments, an antibody (e.g., one antibody or two antibodies) of the present disclosure can be combined with an antibody (e.g., one antibody or two antibodies) described in U.S. Pat. No. 10,787,501 (the '501 patent). In certain embodiments, mAb10987 of the '501 patent is administered in combination with mAb14256 and/or mAb15151 of the present disclosure. In certain embodiments, mAb10933 and mAb10987 of the '501 patent are administered in combination with mAb14256 of the present disclosure. In certain embodiments. mAb10985 and mAb10987 of the '501 patent are administered in combination with mAb15151 of the present disclosure. In certain embodiments, mAb10985 of the '501 patent is administered in combination with mAb14315 and/or mAb15151 of the present disclosure. In certain embodiments, mAb15160 of the present disclosure is administered in combination with mAb10987 of the '501 patent. In certain embodiments, mAb15160 of the present disclosure is administered in combination with mAb10985 of the '501 patent. In certain embodiments, mAb15160 of the present disclosure is administered in combination with mAb10985 and mAb10987 of the '501 patent. In certain embodiments, mAb15160 of the present disclosure is administered in combination with any one, two, or three of mAb14256, mAb14315, and mAb15151, optionally further administered with mAb10987 and/or mAb10985. In certain embodiments, any one of, two of, three of, of four of mAb14256, mAb14315, mAb15151, and mAb15160 is administered in combination with i) mAb10933, ii) mAb10987, or iii) mAb10933 and mAb10987. In certain embodiments, any combination of one, two, three, four, five, six, or seven of mAb10933, mAb10987, mAb10985, mAb14256, mAb1435, mAb15151, and mAb15160 is administered in combination. In certain embodiments, an antibody or antigen-binding fragment thereof selected from mAb10987. mAb14284, mAb14315 and mAb17090 is combined with an antibody or antigen-binding fragment thereof selected from mAb10933, mAb14256, mAb15160 and mAb15160_2. In certain embodiments, the combination comprises mAb10987 and mAb10933. In certain embodiments, the combination comprises mAb10987 and mAb14256. In certain embodiments, the combination comprises mAb10987 and mAb15160. In certain embodiments, the combination comprises mAb10987 and mAb15160_2. In certain embodiments, the combination comprises mAb14284 and mAb10933. In certain embodiments, the combination comprises mAb14284 and mAb14256. In certain embodiments, the combination comprises mAb14284 and mAb15160. In certain embodiments, the combination comprises mAb14284 and mAb15160_2. In certain embodiments, the combination comprises mAb14315 and mAb10933. In certain embodiments, the combination comprises mAb14315 and mAb14256. In certain embodiments, the combination comprises mAb14315 and mAb15160. In certain embodiments the combination comprises mAb14315 and mAbA1560_2. In certain embodiments, the combination comprises mAb17090 and mAb10933. In certain embodiments the combination comprises mAb17090 and mAb14256. In certain embodiments, the combination comprises mAb17090 and mAb160. In certain embodiments, the combination comprises mAb17090 and mAb15160_2. In any of the combinations discussed above or herein, the antibody or antigen-binding fragment may be defined by the CDRs contained within the HCVR and LCVR sequences identified in Table 4. by the heavy and light chain CDR sequences identified in Table 4, by the HCVR and LCVR sequences identified in Table 4, or by the full-length heavy and light chain sequences identified in Table 4, and each of these specific combinations is encompassed within the present disclosure. Certain exemplary combinations of two antibodies are shown below. In some embodiments, an antibody that specifically binds TMPRSS2 is H1H7017N, as described in International Patent Pub. No. WO/2019/147831.

TABLE-US-00002 TABLE 2 Exemplary Combinations of Two Antibodies mAb/ Combination mAb mAb mAb mAb mAb mAb mAb mAb mAb mAb No. 10933 10985 10987 14256 14315 15151 14284 17090 15160 15160_2 mAb10933 X 1 2 3 4 5 6 7 8 9 mAb10985 10 X 11 12 13 14 15 16 17 18 mAb10987 19 20 X 21 22 23 24 25 26 27 mAb14256 28 29 30 X 31 32 33 34 35 36 mAb14315 37 38 39 40 X 41 42 43 44 45 mAb15151 46 47 48 49 50 X 51 52 53 54 mAb14284 55 56 57 58 59 60 X 61 62 63 mAb17090 64 65 66 67 68 69 70 X 71 72 mAb15160 73 74 75 76 77 78 79 80 X 81 mAb15160_2 82 83 84 85 86 87 88 89 90 X

TABLE-US-00003 TABLE3 ExemplarySequencesfromU.S.Pat.No.10,787,501 Antibody Component Designation Part Sequence SEQIDNO AminoAcids mAb10933 HCVR QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYM 1010 SWIRQAPGKGLEWVSYITYSGSTIYYADSVKGRF TISRDNAKSSLYLQMNSLRAEDTAVYYCARDRGT TMVPFDYWGQGTLVTVSS HCDR1 GFTFSDYY 1012 HCDR2 ITYSGSTI 1014 HCDR3 ARDRGTTMVPFDY 1016 LCVR DIQMTQSPSSLSASVGDRVTITCQASQDITNYLN 1018 WYQQKPGKAPKLLIYAASNLETGVPSRFSGSGSG TDFTFTISGLQPEDIATYYCQQYDNLPLTFGGGT KVEIK LCDR1 QDITNY 1020 LCDR2 AAS 1022 LCDR3 QQYDNLPLT 1024 HC QVQLVESGGGLVKPGGSLRLSCAASGFTFSDYYM 1026 SWIRQAPGKGLEWVSYITYSGSTIYYADSVKGRF TISRDNAKSSLYLQMNSLRAEDTAVYYCARDRGT TMVPFDYWGQGTLVTVSSASTKGPSVFPLAPSSK STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELL GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK SLSLSPGK LC DIQMTQSPSSLSASVGDRVTITCQASQDITNYLN 1028 WYQQKPGKAPKLLIYAASNLETGVPSRFSGSGSG TDFTFTISGLQPEDIATYYCQQYDNLPLTFGGGT KVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLL NNFYPREAKVQWKVDNALQSGNSQESVTEQDSKD STYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGEC NucleicAcids HCVR CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGG 1009 TCAAGCCTGGAGGGTCCCTGAGACTCTCCTGTGC AGCCTCTGGATTCACCTTCAGTGACTACTACATG AGCTGGATCCGCCAGGCTCCAGGGAAGGGGCTGG AGTGGGTTTCATACATTACTTATAGTGGTAGTAC CATATACTACGCAGACTCTGTGAAGGGCCGATTC ACCATCTCCAGGGACAACGCCAAGAGCTCACTGT ATCTGCAAATGAACAGCCTGAGAGCCGAGGACAC GGCCGTGTATTACTGTGCGAGAGATCGCGGTACA ACTATGGTCCCCTTTGACTACTGGGGCCAGGGAA CCCTGGTCACCGTCTCCTCA HCDR1 GGATTCACCTTCAGTGACTACTAC 1011 HCDR2 ATTACTTATAGTGGTAGTACCATA 1013 HCDR3 GCGAGAGATCGCGGTACAACTATGGTCCCCTTTG 1015 ACTAC LCVR GACATCCAGATGACCCAGTCTCCATCCTCCCTGT 1017 CTGCATCTGTAGGAGACAGAGTCACCATCACTTG CCAGGCGAGTCAGGACATTACCAACTATTTAAAT TGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGC TCCTGATCTACGCTGCATCCAATTTGGAAACAGG GGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGG ACAGATTTTACTTTCACCATCAGCGGCCTGCAGC CTGAAGATATTGCAACATATTACTGTCAACAGTA TGATAATCTCCCTCTCACTTTCGGCGGAGGGACC AAGGTGGAGATCAAA LCDR1 CAGGACATTACCAACTAT 1019 LCDR2 GCTGCATCC 1021 LCDR3 CAACAGTATGATAATCTCCCTCTCACT 1023 HC CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGG 1025 TCAAGCCTGGAGGGTCCCTGAGACTCTCCTGTGC AGCCTCTGGATTCACCTTCAGTGACTACTACATG AGCTGGATCCGCCAGGCTCCAGGGAAGGGGCTGG AGTGGGTTTCATACATTACTTATAGTGGTAGTAC CATATACTACGCAGACTCTGTGAAGGGCCGATTC ACCATCTCCAGGGACAACGCCAAGAGCTCACTGT ATCTGCAAATGAACAGCCTGAGAGCCGAGGACAC GGCCGTGTATTACTGTGCGAGAGATCGCGGTACA ACTATGGTCCCCTTTGACTACTGGGGCCAGGGAA CCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGG CCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAG AGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCC TGGTCAAGGACTACTTCCCCGAACCGGTGACGGT GTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTG CACACCTTCCCGGCTGTCCTACAGTCCTCAGGAC TCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTC CAGCAGCTTGGGCACCCAGACCTACATCTGCAAC GTGAATCACAAGCCCAGCAACACCAAGGTGGACA AGAAAGTTGAGCCCAAATCTTGTGACAAAACTCA CACATGCCCACCGTGCCCAGCACCTGAACTCCTG GGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAAC CCAAGGACACCCTCATGATCTCCCGGACCCCTGA GGTCACATGCGTGGTGGTGGACGTGAGCCACGAA GACCCTGAGGTCAAGTTCAACTGGTACGTGGACG GCGTGGAGGTGCATAATGCCAAGACAAAGCCGCG GGAGGAGCAGTACAACAGCACGTACCGTGTGGTC AGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAA AGCCCTCCCAGCCCCCATCGAGAAAACCATCTCC AAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGT ACACCCTGCCCCCATCCCGGGATGAGCTGACCAA GAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGC TTCTATCCCAGCGACATCGCCGTGGAGTGGGAGA GCAATGGGCAGCCGGAGAACAACTACAAGACCAC GCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTC CTCTACAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCGTGAT GCATGAGGCTCTGCACAACCACTACACGCAGAAG TCCCTCTCCCTGTCTCCGGGTAAATGA LC GACATCCAGATGACCCAGTCTCCATCCTCCCTGT 1027 CTGCATCTGTAGGAGACAGAGTCACCATCACTTG CCAGGCGAGTCAGGACATTACCAACTATTTAAAT TGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGC TCCTGATCTACGCTGCATCCAATTTGGAAACAGG GGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGG ACAGATTTTACTTTCACCATCAGCGGCCTGCAGC CTGAAGATATTGCAACATATTACTGTCAACAGTA TGATAATCTCCCTCTCACTTTCGGCGGAGGGACC AAGGTGGAGATCAAACGAACTGTGGCTGCACCAT CTGTCTTCATCTTCCCGCCATCTGATGAGCAGTT GAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTG AATAACTTCTATCCCAGAGAGGCCAAAGTACAGT GGAAGGTGGATAACGCCCTCCAATCGGGTAACTC CCAGGAGAGTGTCACAGAGCAGGACAGCAAGGAC AGCACCTACAGCCTCAGCAGCACCCTGACGCTGA GCAAAGCAGACTACGAGAAACACAAAGTCTACGC CTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCC GTCACAAAGAGCTTCAACAGGGGAGAGTGTTAG AminoAcids mAb10987 HCVR QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYAM 1030 YWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRF TISRDNSKNTLYLQMNSLRTEDTAVYYCASGSDY GDYLLVYWGQGTLVTVSS HCDR1 GFTFSNYA 1032 HCDR2 ISYDGSNK 1034 HCDR3 ASGSDYGDYLLVY 1036 LCVR QSALTQPASVSGSPGQSITISCTGTSSDVGGYNY 1038 VSWYQQHPGKAPKLMIYDVSKRPSGVSNRFSGSK SGNTASLTISGLQSEDEADYYCNSLTSISTWVFG GGTKLTVL LCDR1 SSDVGGYNY 1040 LCDR2 DVS 1042 LCDR3 NSLTSISTWV 1044 HC QVQLVESGGGVVQPGRSLRLSCAASGFTFSNYAM 1046 YWVRQAPGKGLEWVAVISYDGSNKYYADSVKGRF TISRDNSKNTLYLQMNSLRTEDTAVYYCASGSDY GDYLLVYWGQGTLVTVSSASTKGPSVFPLAPSSK STSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICN VNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELL GGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVV SVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTIS KAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFF LYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQK SLSLSPGK LC QSALTQPASVSGSPGQSITISCTGTSSDVGGYNY 1048 VSWYQQHPGKAPKLMIYDVSKRPSGVSNRFSGSK SGNTASLTISGLQSEDEADYYCNSLTSISTWVFG GGTKLTVLGQPKAAPSVTLFPPSSEELQANKATL VCLISDFYPGAVTVAWKADSSPVKAGVETTTPSK QSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGS TVEKTVAPTECS NucleicAcids HCVR CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGG 1029 TCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGC AGCCTCTGGATTCACCTTCAGTAACTATGCTATG TACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATCATATGATGGAAGTAA TAAATACTATGCAGACTCCGTGAAGGGCCGATTC ACCATCTCCAGAGACAATTCCAAGAACACGCTGT ATCTGCAAATGAACAGCCTGAGAACTGAGGACAC GGCTGTGTATTACTGTGCGAGTGGCTCCGACTAC GGTGACTACTTATTGGTTTACTGGGGCCAGGGAA CCCTGGTCACCGTCTCCTCA HCDR1 GGATTCACCTTCAGTAACTATGCT 1031 HCDR2 ATATCATATGATGGAAGTAATAAA 1033 HCDR3 GCGAGTGGCTCCGACTACGGTGACTACTTATTGG 1035 TTTAC LCVR CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTG 1037 GGTCTCCTGGACAGTCGATCACCATCTCCTGCAC TGGAACCAGCAGTGACGTTGGTGGTTAT7XACTAT GTCTCCTGGTACCAACAACACCCAGGCAAAGCCC CCAAACTCATGATTTATGATGTCAGTAAGCGGCC CTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAG TCTGGCAACACGGCCTCCCTGACCATCTCTGGGC TCCAGTCTGAGGACGAGGCTGATTATTACTGCAA CTCTTTGACAAGCATCAGCACTTGGGTGTTCGGC GGAGGGACCAAGCTGACCGTCCTA LCDR1 AGCAGTGACGTTGGTGGTTATAACTAT 1039 LCDR2 GATGTCAGT 1041 LCDR3 AACTCTTTGACAAGCATCAGCACTTGGGTG 1043 HC CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGG 1045 TCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGC AGCCTCTGGATTCACCTTCAGTAACTATGCTATG TACTGGGTCCGCCAGGCTCCAGGCAAGGGGCTGG AGTGGGTGGCAGTTATATCATATGATGGAAGTAA TAAATACTATGCAGACTCCGTGAAGGGCCGATTC ACCATCTCCAGAGACAATTCCAAGAACACGCTGT ATCTGCAAATGAACAGCCTGAGAACTGAGGACAC GGCTGTGTATTACTGTGCGAGTGGCTCCGACTAC GGTGACTACTTATTGGTTTACTGGGGCCAGGGAA CCCTGGTCACCGTCTCCTCAGCCTCCACCAAGGG CCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAG AGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCC TGGTCAAGGACTACTTCCCCGAACCGGTGACGGT GTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTG CACACCTTCCCGGCTGTCCTACAGTCCTCAGGAC TCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTC CAGCAGCTTGGGCACCCAGACCTACATCTGCAAC GTGAATCACAAGCCCAGCAACACCAAGGTGGACA AGAAAGTTGAGCCCAAATCTTGTGACAAAACTCA CACATGCCCACCGTGCCCAGCACCTGAACTCCTG GGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAAC CCAAGGACACCCTCATGATCTCCCGGACCCCTGA GGTCACATGCGTGGTGGTGGACGTGAGCCACGAA GACCCTGAGGTCAAGTTCAACTGGTACGTGGACG GCGTGGAGGTGCATAATGCCAAGACAAAGCCGCG GGAGGAGCAGTACAACAGCACGTACCGTGTGGTC AGCGTCCTCACCGTCCTGCACCAGGACTGGCTGA ATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAA AGCCCTCCCAGCCCCCATCGAGAAAACCATCTCC AAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGT ACACCCTGCCCCCATCCCGGGATGAGCTGACCAA GAACCAGGTCAGCCTGACCTGCCTGGTCTAAGGC TTCTATCCCAGCGACATCGCCGTGGAGTGGGAGA GCAATGGGCAGCCGGAGAACAACTACAAGACCAC GCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTC CTCTACAGCAAGCTCACCGTGGACAAGAGCAGGT GGCAGCAGGGGAACGTCTTCTCATGCTCCGTGAT GCATGAGGCTCTGCACAACCACTACACGCAGAAG TCCCTCTCCCTGTCTCCGGGTAAATGA LC CAGTCTGCCCTGACTCAGCCTGCCTCCGTGTCTG 1047 GGTCTCCTGGACAGTCGATCACCATCTCCTGCAC TGGAACCAGCAGTGACGTTGGTGGTTATAACTAT GTCTCCTGGTACCAACAACACCCAGGCAAAGCCC CCAAACTCATGATTTATGATGTCAGTAAGCGGCC CTCAGGGGTTTCTAATCGCTTCTCTGGCTCCAAG TCTGGCAACACGGCCTCCCTGACCATCTCTGGGC TCCAGTCTGAGGACGAGGCTGATTATTACTGCAA CTCTTTGACAAGCATCAGCACTTGGGTGTTCGGC GGAGGGACCAAGCTGACCGTCCTAGGCCAGCCCA AGGCCGCCCCCTCCGTGACCCTGTTCCCCCCCTC CTCCGAGGAGCTGCAGGCCAACAAGGCCACCCTG GTGTGCCTGATCTCCGACTTCTACCCCGGCGCCG TGACCGTGGCCTGGAAGGCCGACTCCTCCCCCGT GAAGGCCGGCGTGGAGACCACCACCCCCTCCAAG CAGTCCAACAACAAGTACGCCGCCTCCTCCTACC TGTCCCTGACCCCCGAGCAGTGGAAGTCCCACCG GTCCTACTCCTGCCAGGTGACCCACGAGGGCTCC ACCGTGGAGAAGACCGTGGCCCCCACCGAGTGCT CCTGA AminoAcids mAb10985 HCVR EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAM 1050 HWVRQAPGKGLEWVSGISWNRGSIGYADSVKGRF TISRDNAKNSLYLQMSSLRAEDTALYYCAKDGER WDSWVPSARNGMDVWGQGTTVTVSS HCDR1 GFTFDDYA 1052 HCDR2 ISWNRGSI 1054 HCDR3 AKDGERWDSVVVPSARNGMDV 1056 LCVR QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYD 1058 VHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSK SGTSASLAITGLQAEDEADYYCQSYDSSLSGSYV FGTGTKVTVL LCDR1 SSNIGAGYD 1060 LCDR2 GNS 1062 LCDR3 QSYDSSLSGSYV 1064 HC EVQLVESGGGLVQPGRSLRLSCAASGFTFDDYAM 1066 HWVRQAPGKGLEWVSGISWNRGSIGYADSVKGRF TISRDNAKNSLYLQMSSLRAEDTALYYCAKDGER WDSVVVPSARNGMDVWGQGTTVTVSSASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCP PCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTC VVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQ YNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALP APIEKTISKAKGQPREPQVYTLPPSRDELTKNQV SLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPV LDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEA LHNHYTQKSLSLSPGK LC QSVLTQPPSVSGAPGQRVTISCTGSSSNIGAGYD 1068 VHWYQQLPGTAPKLLIYGNSNRPSGVPDRFSGSK SGTSASLAITGLQAEDEADYYCQSYDSSLSGSYV FGTGTKVTVLGQPKAAPSVTLFPPSSEELQANKA TLVCLISDFYPGAVTVAWKADSSPVKAGVETTTP SKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECS NucleicAcids HCVR GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGG 1049 TACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGC AGCCTCTGGATTCACCTTTGATGATTATGCCATG CACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGG AGTGGGTCTCAGGTATTAGTTGGAATAGGGGTAG CATAGGCTATGCGGACTCTGTGAAGGGCCGATTC ACCATCTCCAGAGACAACGCCAAGAACTCCCTGT ATCTGCAAATGAGCAGTCTGAGAGCTGAGGACAC GGCCTTGTATTACTGCGCAAAAGATGGCGAGAGA TGGGATAGTGTAGTAGTACCATCTGCTAGGAACG GTATGGACGTCTGGGGCCAAGGGACCACGGTCAC CGTCTCCTCA HCDR1 GGATTCACCTTTGATGATTATGCC 1051 HCDR2 ATTAGTTGGAATAGGGGTAGCATA 1053 HCDR3 GCAAAAGATGGCGAGAGATGGGATAGTGTAGTAG 1055 TACCATCTGCTAGGAACGGTATGGACGTC LCVR CAGTCTGTGCTGACGCAGCCGCCCTCAGTGTCTG 1057 GGGCCCCAGGGCAGAGGGTCACCATCTCCTGCAC TGGGAGCAGCTCCAACATCGGGGCAGGTTATGAT GTACATTGGTACCAGCAGCTTCCAGGAACAGCCC CCAAACTCCTCATCTATGGTAACAGCAATCGGCC CTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAG TCTGGCACCTCAGCCTCCCTGGCCATCACTGGGC TCCAGGCTGAGGATGAGGCTGATTATTACTGCCA GTCCTATGACAGCAGCCTGAGTGGCTCTTATGTC TTCGGAACTGGGACCAAGGTCACCGTCCTA LCDR1 AGCTCCAACATCGGGGCAGGTTATGAT 1059 LCDR2 GGTAACAGC 1061 LCDR3 CAGTCCTATGACAGCAGCCTGAGTGGCTCTTATG 1063 TC HC GAAGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGG 1065 TACAGCCTGGCAGGTCCCTGAGACTCTCCTGTGC AGCCTCTGGATTCACCTTTGATGATTATGCCATG CACTGGGTCCGGCAAGCTCCAGGGAAGGGCCTGG AGTGGGTCTCAGGTATTAGTTGGAATAGGGGTAG CATAGGCTATGCGGACTCTGTGAAGGGCCGATTC ACCATCTCCAGAGACAACGCCAAGAACTCCCTGT ATCTGCAAATGAGCAGTCTGAGAGCTGAGGACAC GGCCTTGTATTACTGCGCAAAAGATGGCGAGAGA TGGGATAGTGTAGTAGTACCATCTGCTAGGAACG GTATGGACGTCTGGGGCCAAGGGACCACGGTCAC CGTCTCCTCAGCCTCCACCAAGGGCCCATCGGTC TTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTG GGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGA CTACTTCCCCGAACCGGTGACGGTGTCGTGGAAC TCAGGCGCCCTGACCAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCAGGACTCTACTCCCT CAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTG GGCACCCAGACCTACATCTGCAACGTGAATCACA AGCCCAGCAACACCAAGGTGGACAAGAAAGTTGA GCCCAAATCTTGTGACAAAACTCACACATGCCCA CCGTGCCCAGCACCTGAACTCCTGGGGGGACCGT CAGTCTTCCTCTTCCCCCCAAAACCCAAGGACAC CCTCATGATCTCCCGGACCCCTGAGGTCACATGC GTGGTGGTGGACGTGAGCCACGAAGACCCTGAGG TCAAGTTCAACTGGTACGTGGACGGCGTGGAGGT GCATAATGCCAAGACAAAGCCGCGGGAGGAGCAG TACAACAGCACGTACCGTGTGGTCAGCGTCCTCA CCGTCCTGCACCAGGACTGGCTGAATGGCAAGGA GTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCA GCCCCCATCGAGAAAACCATCTCCAAAGCCAAAG GGCAGCCCCGAGAACCACAGGTGTACACCCTGCC CCCATCCCGGGATGAGCTGACCAAGAACCAGGTC AGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCA GCGACATCGCCGTGGAGTGGGAGAGCAATGGGCA GCCGGAGAACAACTACAAGACCACGCCTCCCGTG CTGGACTCCGACGGCTCCTTCTTCCTCTACAGCA AGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGG GAACGTCTTCTCATGCTCCGTGATGCATGAGGCT CTGCACAACCACTACACGCAGAAGTCCCTCTCCC TGTCTCCGGGTAAATGA LC CAGTCTGTGCTGACGCAGCCGCCCTCAGTGTCTG 1067 GGGCCCCAGGGCAGAGGGTCACCATCTCCTGCAC TGGGAGCAGCTCCAACATCGGGGCAGGTTATGAT GTACATTGGTACCAGCAGCTTCCAGGAACAGCCC CCAAACTCCTCATCTATGGTAACAGCAATCGGCC CTCAGGGGTCCCTGACCGATTCTCTGGCTCCAAG TCTGGCACCTCAGCCTCCCTGGCCATCACTGGGC TCCAGGCTGAGGATGAGGCTGATTATTACTGCCA GTCCTATGACAGCAGCCTGAGTGGCTCTTATGTC TTCGGAACTGGGACCAAGGTCACCGTCCTAGGCC AGCCCAAGGCCGCCCCCTCCGTGACCCTGTTCCC CCCCTCCTCCGAGGAGCTGCAGGCCAACAAGGCC ACCCTGGTGTGCCTGATCTCCGACTTCTACCCCG GCGCCGTGACCGTGGCCTGGAAGGCCGACTCCTC CCCCGTGAAGGCCGGCGTGGAGACCACCACCCCC TCCAAGCAGTCCAACAACAAGTACGCCGCCTCCT CCTACCTGTCCCTGACCCCCGAGCAGTGGAAGTC CCACCGGTCCTACTCCTGCCAGGTGACCCACGAG GGCTCCACCGTGGAGAAGACCGTGGCCCCCACCG AGTGCTCCTGA

[0228] In some embodiments, anti-CoV-S antigen-binding proteins (e.g., anti-SARS-CoV-2-S antibodies or antigen-binding fragments thereof) from different human donors may be combined. The present invention includes a composition comprising two (or more) anti-SARS-CoV-2-S antibodies or antigen-binding fragments comprising variable domains from human subjects, wherein the two (or more) antibodies or antigen-binding fragments are derived from different subjects (e.g., two different human subjects). Antibody variable regions derived from human B cells are discussed, e.g., in Examples 1 and 2 (Table 6), which describes that variable domains cloned from such B cells are combined with a constant region not from those B cells to produce hybrid antibodies.

[0229] In some embodiments, the further therapeutic agent is an anti-viral drug and/or a vaccine. As used herein, the term anti-viral drug refers to any anti-infective drug or therapy used to treat, prevent, or ameliorate a viral infection in a subject. The term anti-viral drug includes, but is not limited to a cationic steroid antimicrobial, leupeptin, aprotinin, ribavirin, or interferon-alpha2b. Methods for treating or preventing virus (e.g., coronavirus) infection in a subject in need of said treatment or prevention by administering an antibody or antigen-binding fragment of Table 4 in association with a further therapeutic agent are part of the present invention.

[0230] For example, in an embodiment of the invention, the further therapeutic agent is a vaccine, e.g., a coronavirus vaccine. In an embodiment of the invention, a vaccine is an inactivated/killed virus vaccine, a live attenuated virus vaccine or a virus subunit vaccine.

[0231] For example, in an embodiment of the invention, the further therapeutic agent is:

##STR00001##

See Shen et al. Biochimie 142: 1-10 (2017).

[0232] In an embodiment of the invention, the anti-viral drug is an antibody or antigen-binding fragment that binds specifically to coronavirus, e.g., SARS-CoV-2, SARS-CoV, or MERS-CoV. Exemplary anti-CoV-S antibodies include, but are not limited to: H4sH15188P; H1H15188P; H1H15211P; H1H15177P; H4sH15211P; H1H15260P2; H1H15259P2; H1H15203P; H4sH15260P2; H4sH15231P2; H1H15237P2; H1H15208P; H1H15228P2; H1H15233P2; H1H15264P2; H1H15231P2; H1H15253P2; H1H15215P; and H1H15249P2, as set forth in International patent application publication no. WO/2015/179535, or an antigen-binding fragment thereof, e.g., wherein the antibody or fragment comprises a light chain immunoglobulin that includes LCDR1, LCDR2 and LCDR3 (e.g., the V.sub.L or light chain thereof); and a heavy chain that includes HCDR1, HCDR2 and HCDR3 (e.g., the V.sub.H or heavy chain thereof) of any of the foregoing anti-CoV-S antibodies.

[0233] In a certain embodiment of the invention, the further therapeutic agent is not aprotinin, leupeptin, a cationic steroid antimicrobial, an influenza vaccine (e.g., killed, live, attenuated whole virus or subunit vaccine), or an antibody against influenza virus (e.g., an anti-hemagglutinin antibody).

[0234] The term in association with indicates that the components, an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment thereof of the present invention, along with another agent, can be formulated into a single composition, e.g., for simultaneous delivery, or formulated separately into two or more compositions (e.g., a kit). Each component can be administered to a subject at a different time than when the other component is administered; for example, each administration may be given non-simultaneously (e.g., separately or sequentially) at intervals over a given period of time. Moreover, the separate components may be administered to a subject by the same or by a different route (e.g., wherein an anti-CoV-S antibody or antigen-binding fragment thereof.

Kits

[0235] Further provided are kits comprising one or more components that include, but are not limited to, an anti-CoV-S antigen-binding protein, e.g., an antibody or antigen-binding fragment as discussed herein (e.g., of Table 4), in association with one or more additional components including, but not limited to, a further therapeutic agent, as discussed herein. The antigen-binding protein and/or the further therapeutic agent can be formulated as a single composition or separately in two or more compositions, e.g., with a pharmaceutically acceptable carrier, in a pharmaceutical composition.

[0236] In one embodiment of the invention, the kit includes an anti-CoV-S antigen-binding protein, e.g., an antibody or antigen-binding fragment thereof of the invention (e.g., of Table 4), or a pharmaceutical composition thereof in one container (e.g., in a sterile glass or plastic vial) and a further therapeutic agent in another container (e.g., in a sterile glass or plastic vial).

[0237] In another embodiment, the kit comprises a combination of the invention, including an anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment thereof of the invention (e.g., of Table 4), or pharmaceutical composition thereof in combination with one or more further therapeutic agents formulated together, optionally, in a pharmaceutical composition, in a single, common container.

[0238] If the kit includes a pharmaceutical composition for parenteral administration to a subject, the kit can include a device (e.g., an injection device) for performing such administration. For example, the kit can include one or more hypodermic needles or other injection devices as discussed above containing the anti-CoV-S antigen-binding protein, e.g., antibody or antigen-binding fragment thereof of the present invention (e.g., of Table 4).

[0239] The kit can include a package insert including information concerning the pharmaceutical compositions and dosage forms in the kit. Generally, such information aids patients and physicians in using the enclosed pharmaceutical compositions and dosage forms effectively and safely. For example, the following information regarding a combination of the invention may be supplied in the insert: pharmacokinetics, pharmacodynamics, clinical studies, efficacy parameters, indications and usage, contraindications, warnings, precautions, adverse reactions, overdosage, proper dosage and administration, how supplied, proper storage conditions, references, manufacturer/distributor information and patent information.

Diagnostic Uses of the Antibodies

[0240] The anti-CoV-S antigen-binding proteins, e.g., antibodies or antigen-binding fragments thereof of the present invention (e.g., of Table 4), may be used to detect and/or measure CoV-S in a sample. Exemplary assays for CoV-S may include, e.g., contacting a sample with an anti-CoV-S antigen-binding protein of the invention, wherein the anti-CoV-S antigen-binding protein is labeled with a detectable label or reporter molecule or used as a capture ligand to selectively isolate CoV-S from samples. The presence of an anti-CoV-S antigen-binding protein complexed with CoV-S indicates the presence of CoV-S in the sample. Alternatively, an unlabeled anti-CoV-S antibody can be used in combination with a secondary antibody which is itself detectably labeled. The detectable label or reporter molecule can be a radioisotope, such as .sup.3H, .sup.14C, .sup.32P, .sup.35S, or .sup.125I; a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate, or rhodamine; or an enzyme such as alkaline phosphatase, -galactosidase, horseradish peroxidase, or luciferase. Specific exemplary assays that can be used to detect or measure CoV-S in a sample include neutralization assays, enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and fluorescence-activated cell sorting (FACS). Thus, the present invention includes a method for detecting the presence of spike protein polypeptide in a sample comprising contacting the sample with an anti-CoV-S antigen-binding protein and detecting the presence of a CoV-S/anti-CoV-S antigen-binding protein wherein the presence of the complex indicates the presence of CoV-S.

[0241] An anti-CoV-S antigen-binding protein of the invention (e.g., of Table 4) may be used in a Western blot or immune-protein blot procedure for detecting the presence of CoV-S or a fragment thereof in a sample. Such a procedure forms part of the present invention and includes the steps of e.g.: [0242] providing a membrane or other solid substrate comprising a sample to be tested for the presence of CoV-S, e.g., optionally including the step of transferring proteins from a sample to be tested for the presence of CoV-S (e.g., from a PAGE or SDS-PAGE electrophoretic separation of the proteins in the sample) onto a membrane or other solid substrate using a method known in the art (e.g., semi-dry blotting or tank blotting); and contacting the membrane or other solid substrate to be tested for the presence of CoV-S or a fragment thereof with an anti-CoV-S antigen-binding protein of the invention.

[0243] Such a membrane may take the form, for example, of a nitrocellulose or vinyl-based (e.g., polyvinylidene fluoride (PVDF)) membrane to which the proteins to be tested for the presence of CoV-S in a non-denaturing PAGE (polyacrylamide gel electrophoresis) gel or SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) gel have been transferred (e.g., following electrophoretic separation in the gel). Before contacting the membrane with the anti-CoV-S antigen-binding protein, the membrane is optionally blocked, e.g., with non-fat dry milk or the like so as to bind non-specific protein binding sites on the membrane.

[0244] (2) washing the membrane one or more times to remove unbound anti-CoV-S antigen-binding protein and other unbound substances; and [0245] (3) detecting the bound anti-CoV-S antigen-binding protein.

[0246] Detection of the bound antigen-binding protein indicates that the CoV-S protein is present on the membrane or substrate and in the sample. Detection of the bound antigen-binding protein may be by binding the antigen-binding protein with a secondary antibody (an anti-immunoglobulin antibody) which is detectably labeled and, then, detecting the presence of the secondary antibody label.

[0247] The anti-CoV-S antigen-binding proteins (e.g., antibodies and antigen-binding fragments (e.g., of Table 4)) disclosed herein may also be used for immunohistochemistry. Such a method forms part of the present invention and comprises, e.g., [0248] contacting tissue to be tested for the presence of CoV-S protein with an anti-CoV-S antigen-binding protein of the invention; and [0249] detecting the antigen-binding protein on or in the tissue.

[0250] If the antigen-binding protein itself is detectably labeled, it can be detected directly. Alternatively, the antigen-binding protein may be bound by a detectably labeled secondary antibody wherein the label is then detected.

EXAMPLES

[0251] The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the methods and compositions of the invention and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Centigrade, room temperature is about 25 C., and pressure is at or near atmospheric.

Example 1: Generation of Human Antibodies to SARS-CoV-2 Spike Protein (SARS-CoV-2-S)

[0252] Human antibodies to SARS-CoV-2-Spike protein (SARS-CoV-2-S) were generated in a VELOCIMMUNE mouse comprising DNA encoding human immunoglobulin heavy and kappa light chain variable regions or human immunoglobulin heavy and lambda light chain variable regions. Each mouse was immunized with a vector expressing the SARS-CoV-2-S receptor binding domain (RBD) (amino acids 1-1273 of NCBI accession number (MN908947.3), SEQ ID NO: 1008), followed by a booster with a SARS-CoV-2-S vector or a SARS-CoV-2-S protein. The antibody immune response was monitored by a SARS-CoV-2-S-specific immunoassay. Anti-SARS-CoV-2-S antibodies were isolated directly from antigen-positive mouse B cells without fusion to myeloma cells, as described in U.S. Pat. No. 7,582,298, herein specifically incorporated by reference in its entirety. Using this method, fully human anti-SARS-CoV-2-S antibodies (i.e., antibodies possessing human variable domains and human constant domains) were obtained.

[0253] Antibody variable regions were also isolated from human blood samples. Whole blood was received from patients 6-8 weeks after a laboratory-confirmed PCR positive test for SARS-CoV-2 and symptomatic COVID-19 disease. Red blood cells were lysed using an ammonium chloride based lysis buffer (Life Technologies) and B cells were enriched by negative selection. Single B cells that bound the SARS-CoV-2 spike protein were isolated by fluorescent-activated cell sorting (FACS). Isolated B cells were single-well plated and mixed with antibody light and heavy variable region-specific PCR primers. cDNAs for each single B cell were synthesized via a reverse transcriptase (RT) reaction. Each resulting RT product was then split and transferred into two corresponding wells for subsequent antibody heavy and light chain PCRs. One set of the resulting RT products was first amplified by PCR using a 5 degenerate primer specific for antibody heavy variable region leader sequence or a 5 degenerate primer specific for antibody light chain variable region leader sequence and a 3 primer specific for antibody constant region, to form an amplicon. The amplicons were then amplified again by PCR using a 5 degenerate primer specific for antibody heavy variable region framework 1 or a 5 degenerate primer specific for antibody light chain variable region framework 1 and a 3 primer specific for antibody constant region, to generate amplicons for cloning. The antibody heavy chain and light chain derived PCR products were cloned into expression vectors containing heavy constant region and light constant region, respectively, thereby producing expression vectors for hybrid antibodies. The expression vectors expressing full-length heavy and light chain pairs were transfected into CHO cells to produce antibody proteins for testing.

[0254] The biological properties of exemplary antibodies generated in accordance with the methods of this Example are described in detail in the Examples set forth below.

Example 2: Heavy and Light Chain Variable Region Amino Acid and Nucleotide Sequences

[0255] Table 4 sets forth the amino acid sequence identifiers of the heavy and light chain variable regions and CDRs, as well as the heavy chain and light chain sequences, of exemplary anti-SARS-CoV-2-S antibodies. The corresponding nucleic acid sequence identifiers are set forth in Table 5.

TABLE-US-00004 TABLE 4 Amino Acid Sequence Identifiers SEQ ID NOs mAb HCVR HCDR1 HCDR2 HCDR3 LCVR LCDR1 LCDR2 LCDR3 HC LC mAb15163 2 4 6 8 10 12 14 16 18 20 mAb15164 22 24 26 28 30 32 34 36 38 40 mAb15165 42 44 26 47 49 51 34 36 54 56 mAb15166 58 60 62 64 66 51 68 36 70 72 mAb15167 74 76 78 80 82 84 86 88 90 92 mAb15170 94 96 98 100 102 104 106 108 110 112 mAb14296 114 116 118 120 122 124 126 128 130 132 mAb14297 134 136 138 140 142 144 126 146 148 150 mAb14312 152 154 156 158 160 162 164 166 168 170 mAb14313 172 174 176 178 180 182 184 186 188 190 mAb14314 192 194 196 198 200 202 204 206 208 210 mAb14315 212 214 216 218 220 222 126 224 226 228 mAb14316 230 232 234 236 238 240 242 244 246 248 mAb15150 250 252 254 256 258 260 262 264 266 268 mAb15151 270 272 274 276 278 280 106 282 284 286 mAb15156 288 290 292 294 296 298 300 302 304 306 mAb15157 308 310 312 314 316 318 320 322 324 326 mAb15158 328 330 332 334 336 338 106 340 342 344 mAb15159 346 96 98 350 352 354 106 356 358 360 mAb15160 362 364 366 368 370 372 106 374 376 378 mAb15161 380 382 384 386 388 390 392 394 396 398 mAb15162 400 402 98 405 407 409 242 411 413 415 mAb14280 417 419 421 423 425 427 14 429 431 433 mAb14281 435 437 138 440 442 444 446 448 450 452 mAb14282 454 456 458 460 462 84 465 467 469 471 mAb14283 473 475 477 479 481 483 485 487 489 491 mAb14284 493 495 497 499 501 503 505 507 509 511 mAb14285 513 515 517 519 521 523 525 36 527 529 mAb14286 531 533 535 537 539 483 542 544 546 548 mAb14287 550 552 554 556 558 84 14 560 562 564 mAb14288 566 568 570 572 574 576 578 580 582 584 mAb14289 586 588 590 592 594 596 126 598 600 602 mAb14290 604 606 608 610 612 614 126 616 618 620 mAb14291 622 624 626 628 630 632 634 636 638 640 mAb14292 642 644 646 648 650 652 634 655 657 659 mAb14293 661 663 665 667 669 124 126 671 673 675 mAb14295 677 679 78 682 684 686 126 688 690 692 mAb13459 694 696 698 700 702 704 706 708 710 712 mAb14230 714 696 716 718 720 722 126 724 726 728 mAb14231 730 732 734 736 738 740 106 742 744 746 mAb14232 748 750 497 752 754 756 505 758 760 762 mAb14233 764 766 768 770 772 774 776 778 780 782 mAb14234 784 786 788 790 792 794 796 798 800 802 mAb14235 804 806 497 808 810 812 505 814 816 818 mAb14247 820 822 497 825 827 756 14 829 831 833 mAb14248 835 837 839 841 843 845 847 849 851 853 mAb14249 855 857 859 861 863 865 106 867 869 871 mAb14255 873 76 876 878 880 84 86 36 883 885 mAb14256 887 889 891 893 895 897 164 899 901 903 mAb14257 905 154 908 910 912 914 916 166 918 920 mAb14258 922 924 926 928 930 576 933 935 937 939 mAb14259 941 943 945 947 949 951 634 655 953 955 mAb14260 957 959 961 963 965 967 969 971 973 975 mAb13457 694 696 698 700 977 704 706 979 710 981 mAb13458 694 696 698 700 983 704 706 985 710 987 mAb14294 989 991 993 995 997 999 1001 1003 1005 1007 mAb17090 493 495 497 499 501 503 505 507 1075 511 mAb15160_2 362 364 366 368 370 372 106 374 1077 378

TABLE-US-00005 TABLE 5 Nucleic Acid Sequence Identifiers SEQ ID NOs mAb HCVR HCDR1 HCDR2 HCDR3 LCVR LCDR1 LCDR2 LCDR3 HC LC mAb15163 1 3 5 7 9 11 13 15 17 19 mAb15164 21 23 25 27 29 31 33 35 37 39 mAb15165 41 43 45 46 48 50 33 52 53 55 mAb15166 57 59 61 63 65 50 67 52 69 71 mAb15167 73 75 77 79 81 83 85 87 89 91 mAb15170 93 95 97 99 101 103 105 107 109 111 mAb14296 113 115 117 119 121 123 125 127 129 131 mAb14297 133 135 137 139 141 143 125 145 147 149 mAb14312 151 153 155 157 159 161 163 165 167 169 mAb14313 171 173 175 177 179 181 183 185 187 189 mAb14314 191 193 195 197 199 201 203 205 207 209 mAb14315 211 213 215 217 219 221 125 223 225 227 mAb14316 229 231 233 235 237 239 241 243 245 247 mAb15150 249 251 253 255 257 259 261 263 265 267 mAb15151 269 271 273 275 277 279 105 281 283 285 mAb15156 287 289 291 293 295 297 299 301 303 305 mAb15157 307 309 311 313 315 317 319 321 323 325 mAb15158 327 329 331 333 335 337 105 339 341 343 mAb15159 345 347 348 349 351 353 105 355 357 359 mAb15160 361 363 365 367 369 371 105 373 375 377 mAb15161 379 381 383 385 387 389 391 393 395 397 mAb15162 399 401 403 404 406 408 241 410 412 414 mAb14280 416 418 420 422 424 426 13 428 430 432 mAb14281 434 436 438 439 441 443 445 447 449 451 mAb14282 453 455 457 459 461 463 464 466 468 470 mAb14283 472 474 476 478 480 482 484 486 488 490 mAb14284 492 494 496 498 500 502 504 506 508 510 mAb14285 512 514 516 518 520 522 524 52 526 528 mAb14286 530 532 534 536 538 540 541 543 545 547 mAb14287 549 551 553 555 557 83 13 559 561 563 mAb14288 565 567 569 571 573 575 577 579 581 583 mAb14289 585 587 589 591 593 595 125 597 599 601 mAb14290 603 605 607 609 611 613 125 615 617 619 mAb14291 621 623 625 627 629 631 633 635 637 639 mAb14292 641 643 645 647 649 651 653 654 656 658 mAb14293 660 662 664 666 668 123 125 670 672 674 mAb14295 676 678 680 681 683 685 125 687 689 691 mAb13459 693 695 697 699 701 703 705 707 709 711 mAb14230 713 695 715 717 719 721 125 723 725 727 mAb14231 729 731 733 735 737 739 105 741 743 745 mAb14232 747 749 496 751 753 755 504 757 759 761 mAb14233 763 765 767 769 771 773 775 777 779 781 mAb14234 783 785 787 789 791 793 795 797 799 801 mAb14235 803 805 496 807 809 811 504 813 815 817 mAb14247 819 821 823 824 826 755 13 828 830 832 mAb14248 834 836 838 840 842 844 846 848 850 852 mAb14249 854 856 858 860 862 864 105 866 868 870 mAb14255 872 874 875 877 879 83 85 881 882 884 mAb14256 886 888 890 892 894 896 163 898 900 902 mAb14257 904 906 907 909 911 913 915 165 917 919 mAb14258 921 923 925 927 929 931 932 934 936 938 mAb14259 940 942 944 946 948 950 633 654 952 954 mAb14260 956 958 960 962 964 966 968 970 972 974 mAb13457 693 695 697 699 976 703 705 978 709 980 mAb13458 693 695 697 699 982 703 705 984 709 986 mAb14294 988 990 992 994 996 998 1000 1002 1004 1006 mAb17090 492 494 496 498 500 502 504 506 1074 510 mAb15160_2 361 363 365 367 369 371 105 373 1076 377

[0256] Antibodies disclosed herein have fully human variable regions but can have mouse constant regions (e.g., a mouse IgG1 Fc or a mouse IgG2 Fc (a or b isotype)) or human constant regions (e.g., a human IgG1 Fc or a human IgG4 Fc). As will be appreciated by a person of ordinary skill in the art, an antibody having a particular Fc isotype can be converted to an antibody with a different Fc isotype (e.g., an antibody with a mouse IgG1 Fc can be converted to an antibody with a human IgG4, etc.), but in any event, the variable domains (including the CDRs)which are indicated by the numerical identifiers shown in Tables 4 and 5 will remain the same, and the binding properties to antigen are expected to be identical or substantially similar regardless of the nature of the constant domain.

[0257] As described above, the antibodies were obtained by direct isolation from antigen-positive VELOCIMMUNE mouse B cells or derived from variable regions cloned from antigen-positive human B cells. A summary of these sources is shown in Table 6.

TABLE-US-00006 TABLE 6 Antibody/Variable Region sources mAb Source mAb13457 mouse B cells mAb13458 mouse B cells mAb13459 mouse B cells mAb14230 human B cells mAb14231 human B cells mAb14232 human B cells mAb14233 human B cells mAb14234 human B cells mAb14235 human B cells mAb14247 human B cells mAb14248 human B cells mAb14249 human B cells mAb14255 mouse B cells mAb14256 mouse B cells mAb14257 mouse B cells mAb14258 mouse B cells mAb14259 human B cells mAb14260 mouse B cells mAb14280 human B cells mAb14281 human B cells mAb14282 human B cells mAb14283 human B cells mAb14284 human B cells mAb14285 human B cells mAb14286 human B cells mAb14287 human B cells mAb14288 human B cells mAb14290 human B cells mAb14289 human B cells mAb14291 human B cells mAb14292 human B cells mAb14293 human B cells mAb14294 human B cells mAb14295 human B cells mAb14296 human B cells mAb14297 human B cells mAb14312 mouse B cells mAb14313 mouse B cells mAb14314 mouse B cells mAb14315 human B cells mAb14316 human B cells mAb15156 mouse B cells mAb15157 mouse B cells mAb15158 human B cells mAb15159 human B cells mAb15160 human B cells mAb15161 human B cells mAb15162 human B cells mAb15163 human B cells mAb15164 human B cells mAb15165 human B cells mAb15166 human B cells mAb15167 human B cells mAb15170 human B cells mAb15150 human B cells mAb15151 human B cells

Example 3: Biacore Binding Kinetics of Purified Anti-SARS-CoV-2-S Monoclonal Antibodies

[0258] Equilibrium dissociation constant (K.sub.D) for different SARS-COV-2 RBD reagents binding to purified CHOt anti-SARS-COV-2 monoclonal antibodies (mAbs) were determined using a real-time surface plasmon resonance based Biacore T200/Biacore 8K biosensor. All binding studies were performed in 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, and 0.05% v/v Surfactant Tween-20, pH 7.4 (HBS-ET) running buffer at 25 C. and 37 C. The Biacore CM5 sensor chip surface was first derivatized by amine coupling with either mouse anti-human Fc specific mAb (Regeneron, mAb2567) to capture anti-SARS-COV-2 mAbs. Binding studies were performed on human SARS-COV-2 RBD extracellular domain expressed with a C-terminal myc-myc-hexahistidine tag (SARS-COV-2 RBD-mmH) (monomeric RBD) (SEQ ID NO: 1069), SARS-COV-2 RBD extracellular domain expressed with a C-terminal mouse IgG2a Fc (SARS-COV-2 RBD mFc) (dimeric RBD) (SEQ ID NO: 1070), and SARS-CoV2 Spike ecto foldon Trimer expressed with a C-terminal myc-myc-hexahistidine (SARS-CoV2 Spike ECD foldon) (trimeric RBD). Use of these reagents allowed for the testing of the antibodies' ability to bind monomeric, dimeric, and trimeric RBD peptides, respectively.

[0259] Three concentrations, 50 nM, 12.5 nM and 3.12 nM, of hSARS-COV-2 RBD-mmH, SARS-COV-2 RBD mFc and SARS-CoV2 Spike ECD foldon prepared in HBS-ET running buffer, were injected for 1.5-3 minutes at a flow rate of 50 L/min while the dissociation of mAb bound different SARS-COV-2 RBD reagents was monitored for 5-8 minutes in HBS-ET running buffer. At the end of each cycle, the SARS-COV-2 RBD mAb capture surface was regenerated using either 12 sec injection of 20 mM phosphoric acid for mouse anti-human Fc specific mAb surface. The association rate (k.sub.a) and dissociation rate (k.sub.d) were determined by fitting the real-time binding sensorgrams to a 1:1 binding model with mass transport limitation using BiaEvaluation software v3.1 or Biacore Insight Evaluation software v2.0. or curve-fitting software. Binding dissociation equilibrium constant (K.sub.D) and dissociative half-life (t) were calculated from the kinetic rates as:

[00001]$K_D\left(M\right)=\frac{\mathrm{kd}}{\mathrm{ka}},\mathrm{and}{t}^{1/2}\left(\min \right)=\frac{\ln \left(2\right)}{60*\mathrm{kd}}$

[0260] Binding kinetics parameters for different anti-SARS-COV-2 mAbs binding to monomeric, dimeric, and trimeric SARS-COV-2 RBD reagents of the invention at 25 C. and 37 C. are shown in Tables 7 through 12, respectively. An isotype control, mAb1932, was also used as a control.

TABLE-US-00007 TABLE 7 Kinetic Binding Parameters of the Interaction of Anti-SARS-COV-2 Monoclonal Antibodies to Monomeric SARS-COV-2 RBD-mmH (SEQ ID NO: 1069) at 25 C. RBD mAb monomer Capture bound at Level 50 nM k.sub.a k.sub.d KD t mAb Captured (RU) (RU) (1/Ms) (1/s) (M) (min) mAb14312 468.6 4.6 190.8 1.53E+06 9.29E04 6.06E10 12.4 mAb14260 415.3 1.6 173.5 1.87E+06 1.38E03 7.38E10 8.4 mAb14284 731.1 1.2 236.5 6.66E+05 5.49E04 8.24E10 21.0 mAb14258 423.9 0.4 180.2 1.89E+06 1.61E03 8.54E10 7.2 mAb14294 408.7 2.0 138.3 6.00E+05 5.48E04 9.14E10 21.1 mAb14283 410.1 8.9 171.9 1.46E+06 1.58E03 1.08E09 7.3 mAb14235 340.0 4.0 117.1 7.89E+05 1.05E03 1.33E09 11.0 mAb14289 395.1 1.1 151 1.16E+06 1.58E03 1.36E09 7.3 mAb13458 435.7 4.4 158.4 9.09E+05 1.39E03 1.53E09 8.3 mAb14313 532.2 1.9 218.2 1.42E+06 2.17E03 1.53E09 5.3 mAb14247 179.0 0.5 63.3 8.38E+05 1.63E03 1.94E09 7.1 mAb14315 465.8 0.9 127.1 4.07E+05 1.02E03 2.49E09 11.4 mAb13459 665.7 4.5 219.3 6.93E+05 1.74E03 2.51E09 6.6 mAb13457 412.5 7.4 148.4 7.74E+05 2.05E03 2.65E09 5.6 mAb14257 518.6 1.0 215.1 3.40E+06 9.52E03 2.80E09 1.2 mAb14255 432.4 4.8 180.4 1.75E+06 4.99E03 2.84E09 2.3 mAb14286 394.6 5.2 158.5 3.23E+06 1.04E02 3.22E09 1.1 mAb14282 414.4 4.7 170.1 1.93E+06 9.57E03 4.97E09 1.2 mAb14281 423.6 0.4 157.8 1.10E+06 5.54E03 5.02E09 2.1 mAb14230 315.0 0.3 113.6 1.13E+06 8.52E03 7.51E09 1.4 mAb14256 440.5 3.6 175.4 1.11E+06 9.27E03 8.32E09 1.2 mAb14285 443.1 0.6 157.9 1.80E+06 2.11E02 1.17E08 0.5 mAb14280 474.3 0.5 177.3 1.66E+06 2.55E02 1.54E08 0.5 mAb14234 458.8 0.9 152.8 5.50E+05 1.13E02 2.05E08 1.0 mAb14232 71.6 1.4 18.8 1.12E+06 3.50E02 3.12E08 0.3 mAb14292 431.3 0.7 11.1 4.17E+05 1.71E02 4.09E08 0.7 mAb14314 597.8 0.8 11.6 IC IC IC IC mAb14248 400.8 1.6 269.5 7.06E+05 2.05E05 2.90E11 564.0 mAb14295 441.5 3.7 59.8 3.01E+05 <1.00E05 3.32E11 21155.0 mAb14233 492.8 3.4 85.7 4.94E+05 3.6E05 7.41E11 315.5 mAb14287 419.8 0.3 91.9 3.10E+05 4.59E05 1.48E10 251.4 mAb14293 432.5 0.6 119.8 2.43E+05 8.10E05 3.33E10 142.6 mAb14291 391.3 3.3 47.8 IC IC IC IC mAb14249 407.5 1.0 42.3 IC IC IC IC mAb14316 264.7 1.6 40 IC IC IC IC mAb14296 529.6 0.7 32.3 IC IC IC IC mAb14231 407.0 2.2 27.3 IC IC IC IC mAb14288 458.5 2.3 24.7 IC IC IC IC mAb14297 444.0 1.4 13.7 IC IC IC IC mAb14290 378.1 1.2 12 IC IC IC IC mAb14259 335.8 2.8 7.8 IC IC IC IC mAb15156 427.3 17.1 91 2.89E+05 1.21E03 4.18E09 9.6 mAb15157 291.6 11.5 20.3 2.78E+05 1.25E03 4.49E09 9.3 mAb15158 317.3 6.6 26.3 1.04E+05 8.12E05 7.79E10 142.2 mAb15159 286.3 16.0 62 4.89E+04 1.30E04 2.66E09 88.8 mAb15160 304.2 8.3 9.3 IC IC IC IC mAb15161 250.6 15.3 40.3 1.73E+05 2.34E04 1.35E09 49.3 mAb15162 287.1 12.2 49.6 1.16E+05 8.50E04 7.30E09 13.6 mAb15163 358.2 12.8 51 2.18E+05 1.67E05 7.79E11 693.3 mAb15164 315.8 11.1 41.2 2.28E+05 3.49E04 1.53E09 33.1 mAb15165 271.1 13.8 33.9 5.39E+05 2.12E04 3.93E10 54.6 mAb15166 249.2 11.0 13.3 2.46E+05 5.35E03 2.17E08 2.2 mAb15167 329.5 10.9 21.8 2.65E+05 4.14E03 1.56E08 2.8 mAb15170 278.8 9.9 21.8 2.00E+05 1.53E05 1.00E10 756.4 mAb15150 276.5 9.6 33.3 7.74E+05 4.61E05 5.94E11 250.8 mAb15151 354.8 13.2 63.4 2.00E+05 7.78E04 3.89E09 14.8 mAb1932 790.6 9.6 0.6 NB NB NB NB NB: No binding (NB) was observed under the current experimental conditions. IC: Observed binding did not fit to the binding simulation model and no binding kinetic parameters were determined under the current experimental conditions. 1.00E05 indicates that no dissociation was observed under the current experimental conditions and the k.sub.d value was manually fixed at 1.00E05 s.sup.1 while fitting the real time binding sensorgrams.

TABLE-US-00008 TABLE 8 Kinetic Binding Parameters of the Interaction of Anti-SARS-COV-2 Monoclonal Antibodies to Monomeric SARS-COV-2 RBD-mmH (SEQ ID NO: 1069) at 37 C. RBD mAb monomer Capture bound at mAb Level 50 nM k.sub.a k.sub.d KD t Captured (RU) (RU) (1/Ms) (1/s) (M) (min) mAb14312 543 221.2 1.57E+06 9.88E04 6.31E10 11.7 mAb14260 525.5 219.6 1.95E+06 1.44E03 7.41E10 8.0 mAb14284 867.3 345.3 6.44E+05 6.05E04 9.40E10 19.1 mAb14258 521 215.5 2.14E+06 1.70E03 7.96E10 6.8 mAb14294 465.5 185.9 6.00E+05 5.90E04 9.84E10 19.6 mAb14283 531.3 217.1 1.54E+06 1.79E03 1.17E09 6.4 mAb14235 388.4 153.8 7.93E+05 1.09E03 1.37E09 10.6 mAb14289 461.9 185.8 1.20E+06 1.76E03 1.46E09 6.5 mAb13458 550.6 212.8 9.07E+05 1.39E03 1.54E09 8.3 mAb14313 676 272.8 2.81E+06 2.59E03 9.21E10 4.5 mAb14247 222.1 91.2 8.32E+05 1.63E03 1.95E09 7.1 mAb14315 581.1 193 4.35E+05 9.85E04 2.26E09 11.7 mAb13459 822.4 299 7.86E+05 2.04E03 2.60E09 5.7 mAb13457 472.1 180.3 8.01E+05 2.34E03 2.92E09 4.9 mAb14257 600.6 242.3 3.26E+06 7.62E03 2.34E09 1.5 mAb14255 518.4 206 1.94E+06 5.35E03 2.76E09 2.2 mAb14286 436.6 169.1 3.38E+06 1.04E02 3.07E09 1.1 mAb14282 492.7 186.2 1.98E+06 1.06E02 5.37E09 1.1 mAb14281 486.7 182 1.12E+06 5.82E03 5.20E09 2.0 mAb14230 369.8 133.2 1.15E+06 8.76E03 7.61E09 1.3 mAb14256 555.8 193.4 1.40E+06 1.15E02 8.24E09 1.0 mAb14285 543.5 165.6 2.09E+06 2.74E02 1.31E08 0.4 mAb14280 586 176.2 1.53E+06 2.48E02 1.62E08 0.5 mAb14234 565.9 162 5.34E+05 1.19E02 2.23E08 1.0 mAb14232 98.1 21.1 4.69E+06 3.72E02 7.92E09 0.3 mAb14292 507.6 10.4 3.59E+05 2.18E02 6.07E08 0.5 mAb14314 693.8 12.5 4.79E+05 1.15E02 2.39E08 1.0 mAb14248 509.5 491.7 7.02E+05 1.90E05 2.71E11 607.3 mAb14295 558.8 196.2 5.78E+05 <1.00E05 1.73E11 21155.0 mAb14233 561.8 276.8 3.20E+05 3.38E05 1.06E10 341.7 mAb14287 528.8 154.9 7.04E+05 4.63E05 6.58E11 249.4 mAb14293 531 166.7 9.62E+05 8.69E05 9.03E11 132.9 mAb14291 516.4 224.5 3.08E+05 4.46E05 1.45E10 258.9 mAb14249 482.5 143 2.38E+05 5.34E05 2.25E10 216.5 mAb14316 304.6 132.1 8.05E+05 <1.00E05 1.24E11 21155.0 mAb14296 672.2 91.3 1.33E+05 <1.00E05 7.55E11 21155.0 mAb14231 490.1 54.7 3.18E+05 1.57E04 4.94E10 73.5 mAb14288 576.6 72.9 1.06E+05 5.85E05 5.54E10 197.5 mAb14297 529.3 32.1 3.01E+05 2.09E04 6.94E10 55.3 mAb14290 457.9 32.9 2.34E+05 1.09E05 4.66E11 1057.7 mAb14259 395.2 9 NB NB NB NB mAb15156 463.2 0.7 106.9 3.73E+05 5.35E03 1.44E08 2.2 mAb15157 309.1 1 65.7 3.44E+05 3.99E03 1.16E08 2.9 mAb15158 298 0.5 44.3 1.26E+05 2.07E04 1.65E09 55.7 mAb15159 293.3 1.9 42 1.46E+05 1.76E04 1.21E09 65.6 mAb15160 400.6 1.2 79.3 2.50E+05 1.94E03 7.79E09 5.9 mAb15161 283.9 0.9 61.1 2.63E+05 8.66E04 3.29E09 13.3 mAb15162 322.4 2.9 38.9 1.03E+05 2.99E03 2.92E08 3.9 mAb15163 343.6 1.7 72.7 2.18E+05 3.87E04 1.78E09 29.9 mAb15164 385 1.1 93.4 2.83E+05 1.79E03 6.33E09 6.5 mAb15165 276.1 2.3 82.5 4.75E+05 1.47E03 3.10E09 7.9 mAb15166 266.8 0.3 19.7 3.50E+05 3.34E02 9.53E08 0.3 mAb15167 405.3 0.6 46.8 3.52E+05 1.95E02 5.55E08 0.6 mAb15170 299.4 1.4 56.3 2.83E+05 8.36E05 2.97E10 138.1 mAb15150 248.5 1.2 64.9 4.93E+05 7.84E05 1.58E10 147.4 mAb15151 595.8 1.6 141.4 1.80E+05 4.42E03 2.46E08 2.6 mAb1932 921.8 2 1.3 NB NB NB NB NB: No binding (NB) was observed under the current experimental conditions. IC: Observed binding did not fit to the binding simulation model and no binding kinetic parameters were determined under the current experimental conditions. 1.00E05 indicates that no dissociation was observed under the current experimental conditions and the k.sub.d value was manually fixed at 1.00E05 s.sup.1 while fitting the real time binding sensorgrams.

TABLE-US-00009 TABLE 9 Kinetic Binding Parameters of the Interaction of Anti-SARS-COV-2 Monoclonal Antibodies to Dimeric SARS-COV-2 RBD mFc (SEQ ID NO: 1070) at 25 C. RBD mFc mAb dimer Capture bound Level at k.sub.a k.sub.d KD t mAb Captured (RU) 50 nM (1/Ms) (1/s) (M) (min) mAb14312 465.2 3.8 349.2 1.68E+06 5.60E05 3.34E11 206.4 mAb14260 416.5 2.2 327.4 1.96E+06 8.70E05 4.43E11 132.8 mAb14284 732.9 0.5 482.2 8.31E+05 2.12E05 2.55E11 544.6 mAb14258 420.7 1 332.8 3.56E+04 1.09E04 3.04E09 106.5 mAb14294 404.3 5.6 291.4 9.11E+05 2.68E05 2.94E11 430.3 mAb14283 417.5 0.6 319.6 1.55E+06 8.14E05 5.24E11 141.9 mAb14235 338.6 2.2 242.7 1.10E+06 6.89E05 6.27E11 167.6 mAb14289 393.3 2.8 295.3 1.43E+06 8.95E05 6.28E11 129.1 mAb13458 439.8 2.5 326.7 1.18E+06 7.55E05 6.40E11 153.1 mAb14313 528.5 10.9 407.4 2.36E+06 9.97E05 4.22E11 115.9 mAb14247 177.9 0.7 135.5 1.19E+06 1.56E04 1.31E10 73.9 mAb14315 455.9 5.0 303.7 7.99E+05 6.26E05 7.84E11 184.4 mAb13457 410.1 7 308.2 1.33E+06 8.59E05 6.45E11 134.5 mAb14257 519.6 2.1 403.1 2.81E+06 2.54E04 9.07E11 45.4 mAb14255 437.3 0.4 340.7 2.19E+06 1.59E04 7.26E11 72.6 mAb14286 393.5 1.5 296.8 2.95E+06 6.55E04 2.22E10 17.6 mAb14282 418.3 0.9 326.3 2.17E+06 4.03E04 1.86E10 28.7 mAb14281 424.2 0.6 305.1 1.46E+06 2.13E04 1.46E10 54.3 mAb14230 315.5 0.6 240.6 2.00E+06 2.86E04 1.42E10 40.5 mAb14256 440.9 5.5 331.9 2.84E+06 1.81E04 6.38E11 63.7 mAb14285 435.9 1 318.7 5.78E+06 5.02E04 8.68E11 23 mAb14280 476.4 0.9 358.2 3.02E+06 3.70E04 1.23E10 31.2 mAb14234 458.2 5 328.7 1.02E+06 1.21E04 1.19E10 95.2 mAb14232 71.7 1.4 52.4 1.97E+06 1.73E03 8.78E10 6.7 mAb14292 431.8 1.4 76.8 1.04E+06 3.51E02 3.36E08 0.3 mAb14314 601.8 1.1 173.6 2.22E+06 5.60E02 2.53E08 0.2 mAb14248 404.4 1.8 4.9 NB NB NB NB mAb14295 441.3 4.7 8.4 NB NB NB NB mAb14233 491.0 2.3 2.3 NB NB NB NB mAb14287 424.9 3.8 9.4 NB NB NB NB mAb14293 440.5 0.3 5.8 NB NB NB NB mAb14291 391.0 4.3 3.3 NB NB NB NB mAb14249 412.7 1.1 4.6 NB NB NB NB mAb14316 266.4 1.7 11.4 IC IC IC IC mAb14296 529.8 1.7 4.6 NB NB NB NB mAb14231 407.8 1 4.8 NB NB NB NB mAb14288 462.8 1.2 4 NB NB NB NB mAb14297 443.5 1.2 6.8 NB NB NB NB mAb14290 382.1 3.6 7.7 NB NB NB NB mAb14259 339.7 0.5 8.3 NB NB NB NB mAb15156 914.2 2.1 493.4 1.41E+06 5.22E05 3.69E11 221.2 mAb15157 543.7 0.5 294.3 3.57E+05 6.67E05 1.87E10 173.2 mAb15158 573.9 1.8 175.4 4.53E+05 1.00E05 2.21E11 1,155.0 mAb15159 519.6 0.8 224.5 6.91E+05 1.19E05 1.72E11 971.4 mAb15160 610.1 1.6 268.3 2.58E+05 1.05E05 4.06E11 1,101.0 mAb15161 547.4 0.5 328.4 1.06E+06 1.26E05 1.19E11 913.8 mAb15162 771.1 0.3 354.1 1.29E+06 2.30E04 1.79E10 50.2 mAb15163 529.5 1.1 164.2 9.30E+05 1.00E05 1.08E11 1,155.0 mAb15164 719.6 0.5 308.8 3.72E+05 2.12E05 5.69E11 545.6 mAb15165 644.5 6.2 222.6 3.01E+05 3.88E+05 1.29E10 297.5 mAb15166 715.6 4.0 334.4 1.02E+06 1.62E05 1.60E11 713.0 mAb15167 501.7 2.5 241.5 4.81E+05 1.05E04 2.17E10 110.3 mAb15170 608.7 2.3 237.5 3.39E+05 1.00E05 2.95E11 1,155.0 mAb15151 1143 2.6 491.7 9.24E+06 3.12E05 3.38E12 369.7 mAb1932 1175.9 2.3 6.8 NB NB NB NB NB: No binding (NB) was observed under the current experimental conditions. IC: Observed binding did not fit to the binding simulation model and no binding kinetic parameters were determined under the current experimental conditions.

TABLE-US-00010 TABLE 10 Kinetic Binding Parameters of the Interaction of Anti-SARS-COV-2 Monoclonal Antibodies to dimeric SARS-COV-2 RBD mFc (SEQ ID NO: 1070) at 37 C.. RBD mFc mAb dimer Capture bound mAb Level at k.sub.a k.sub.d KD t Captured (RU) 50 nM (1/Ms) (1/s) (M) (min) mAb14312 550.3 422.3 1.67E+06 5.80E05 3.47E11 199.1 mAb14260 521.7 408.3 1.93E+06 1.04E04 5.40E11 110.6 mAb14284 872 659.3 8.30E+05 2.24E05 2.70E11 516.1 mAb14258 271.5 392.5 IC IC IC IC mAb14294 467.7 370.4 9.06E+05 3.50E05 3.86E11 330.1 mAb14283 534.6 414.9 1.52E+06 9.59E05 6.31E11 120.5 mAb14235 393.8 308.2 1.09E+06 6.68E05 6.12E11 173 mAb14289 467.8 372.3 1.41E+06 1.28E04 9.07E11 90.1 mAb13458 548.4 425.4 1.16E+06 9.12E05 7.84E11 126.7 mAb14313 703.3 538 2.52E+06 1.46E04 5.82E11 78.9 mAb14247 223 191.8 1.18E+06 1.50E04 1.27E10 77 mAb14315 577.7 416.2 8.77E+05 7.91E05 9.03E11 145.9 mAb13459 830 613.1 9.85E+05 9.24E05 9.39E11 125 mAb13457 467.7 357.9 1.26E+06 1.17E04 9.35E11 98.5 mAb14257 597.6 482 2.81E+06 3.39E04 1.21E10 34.1 mAb14255 523 407.4 2.12E+06 2.50E04 1.18E10 46.2 mAb14286 433.2 336.7 2.94E+06 5.28E04 1.80E10 21.9 mAb14282 493.7 393.8 2.21E+06 5.46E04 2.47E10 21.1 mAb14281 481.8 370.1 1.34E+06 3.65E04 2.71E10 31.7 mAb14230 374 294.6 1.82E+06 4.90E04 2.68E10 23.6 mAb14256 573.9 430.4 1.78E+06 4.73E04 2.67E10 24.4 mAb14285 537 386.8 4.45E+06 1.03E03 2.31E10 11.2 mAb14280 589.2 444.9 2.92E+06 7.72E04 2.64E10 15 mAb14234 571.3 414.8 9.80E+05 4.02E04 4.10E10 28.8 mAb14232 103.7 69 2.10E+06 1.88E03 8.93E10 6.1 mAb14292 503.4 97.9 1.86E+06 6.44E02 3.46E08 0.2 mAb14314 700.2 167.3 1.48E+06 3.57E02 2.41E08 0.3 mAb14248 506.9 8.1 NB NB NB NB mAb14295 555.3 13.8 NB NB NB NB mAb14233 559.6 3.9 NB NB NB NB mAb14287 528.2 19.8 NB NB NB NB mAb14293 532.7 9.1 NB NB NB NB mAb14291 529.1 6.1 NB NB NB NB mAb14249 481 6.6 NB NB NB NB mAb14316 303.8 14.5 NB NB NB NB mAb14296 667.7 7.6 NB NB NB NB mAb14231 494.9 8 NB NB NB NB mAb14288 572.7 8.4 NB NB NB NB mAb14297 525.8 12.4 NB NB NB NB mAb14290 450.6 10.3 NB NB NB NB mAb14259 395.4 9.4 NB NB NB NB mAb15156 1206.3 2.9 672.7 1.78E+05 1.12E04 6.30E10 103.2 mAb15157 689.3 1.8 411.9 9.22E+05 1.07E04 1.16E10 107.9 mAb15158 699.6 1.7 279.4 1.71E+05 2.90E05 1.70E10 398.3 mAb15159 756.1 1.1 314.8 2.20E+05 2.99E05 1.35E10 386.3 mAb15160 913.9 1.3 440.3 4.36E+05 7.90E05 1.81E10 146.2 mAb15161 674.7 0.6 352.5 1.05E+06 4.90E05 4.69E11 235.7 mAb15162 835.4 2.9 322.7 2.00E+05 1.01E04 5.00E10 114.8 mAb15163 778.3 3.3 407 4.16E+05 2.50E05 6.01E11 462.0 mAb15164 937.9 2.2 466.3 1.82E+06 9.40E05 5.18E11 122.9 mAb15165 716.1 0 191.1 1.00E+05 4.55E05 4.50E10 254.0 mAb15166 616.1 2.1 350.6 7.81E+05 9.80E05 1.26E10 117.9 mAb15167 993.3 3.2 499.8 1.40E+06 1.24E04 8.90E11 93.1 mAb15170 779.1 2.9 374.2 2.60E+05 2.41E05 9.10E11 480.0 mAb15150 554 1.8 338.7 9.46E+05 2.80E05 2.96E11 412.5 mAb15151 1452 4.1 653.6 3.70E+06 3.10E03 8.50E10 3.7 mAb1932 2129.7 4 20.6 NB NB NB NB NB: No binding (NB) was observed under the current experimental conditions. IC: Observed binding did not fit to the binding simulation model and no binding kinetic parameters were determined under the current experimental conditions.

TABLE-US-00011 TABLE 11 Kinetic Binding Parameters of the Interaction of Anti-SARS-COV-2 Monoclonal Antibodies to Trimeric SARS-CoV2 Spike ECD foldon Fusion Protein at 25 C.. mAb Spike Capture trimer.his Level bound at k.sub.a k.sub.d KD t mAb Captured (RU) 50 nM (1/Ms) (1/s) (M) (min) mAb14312 463.3 4.0 393.4 1.93E+06 5.24E05 2.72E11 220.5 mAb14260 411.6 6.6 378.1 1.52E+06 5.87E05 3.86E11 196.6 mAb14284 729.9 1.8 469.4 8.12E+05 1.31E05 1.61E11 883 mAb14258 420 1.7 406.3 2.12E+06 4.91E05 2.32E11 235.3 mAb14294 400.7 3.8 341.7 6.97E+05 2.29E05 3.29E11 503.7 mAb14283 417.4 5.5 352.5 1.61E+06 8.66E05 5.39E11 133.3 mAb14235 336.9 1.2 297.5 7.79E+05 3.69E05 4.74E11 312.9 mAb14289 387.0 3.9 345.3 9.12E+05 6.35E05 6.96E11 182 mAb13458 436.2 3.7 340.8 1.21E+06 4.26E05 3.51E11 271.3 mAb14313 528.7 2.2 462 2.00E+06 1.13E04 5.65E11 102.1 mAb14247 172 4.9 143.3 9.44E+05 4.91E05 5.21E11 235 mAb14315 458.4 1.1 335.4 6.52E+05 3.15E05 4.83E11 366.2 mAb13459 668.9 8.1 483.2 1.41E+06 5.46E05 3.89E11 211.5 mAb13457 405.4 4.6 358.2 7.68E+05 5.52E05 7.18E11 209.2 mAb14257 518.4 1.3 535.3 2.45E+06 1.07E04 4.34E11 108.5 mAb14255 431.9 3.2 454.1 1.96E+06 1.38E04 7.06E11 83.5 mAb14286 389.5 2.0 425 2.63E+06 8.71E05 3.31E11 132.6 mAb14282 413.2 3.4 421.7 2.14E+06 1.33E04 6.24E11 86.6 mAb14281 415 5.6 310.7 8.34E+05 8.09E05 9.70E11 142.8 mAb14230 313.1 2.6 340 1.51E+06 6.09E05 4.04E11 189.8 mAb14256 441.9 6.3 356.7 1.47E+06 1.89E04 1.29E10 61 mAb14285 435.7 1.6 486.2 2.34E+06 3.98E05 1.70E11 290.4 mAb14280 472.8 1.3 458.7 1.99E+06 2.69E05 1.36E11 428.7 mAb14234 455.7 5.8 365.5 7.07E+05 5.48E05 7.75E11 210.7 mAb14232 69.7 1 72.8 7.63E+05 1.49E04 1.95E10 77.7 mAb14292 429.2 0.5 138.4 5.84E+05 4.21E03 7.21E09 2.7 mAb14314 600.4 1.7 184 4.92E+05 4.21E03 8.56E09 2.7 mAb14248 403.3 0.4 1.4 NB NB NB NB mAb14295 446.7 2.3 2.4 NB NB NB NB mAb14233 488.5 2.0 0.3 NB NB NB NB mAb14287 421.9 3.0 0.9 NB NB NB NB mAb14293 448.8 0.4 2.4 NB NB NB NB mAb14291 396.1 4.2 0.3 NB NB NB NB mAb14249 413.5 1.6 0.1 NB NB NB NB mAb14316 269.1 1.4 3.9 NB NB NB NB mAb14296 528.3 1.5 1.1 NB NB NB NB mAb14231 410.9 1.7 0.2 NB NB NB NB mAb14288 459.0 0.8 1.9 NB NB NB NB mAb14297 451.0 0.8 1.4 NB NB NB NB mAb14290 379.4 6.1 3.8 NB NB NB NB mAb14259 340.6 0.2 1.9 NB NB NB NB mAb15156 447.5 29.5 204.7 3.42E+05 6.61E05 1.93E10 174.6 mAb15157 303.7 13.3 42.5 3.02E+05 9.28E05 3.08E10 124.5 mAb15158 328.2 11.9 55.1 9.15E+04 1.40E05 1.53E10 825.0 mAb15159 307.7 25.7 147.1 8.75E+04 2.02E05 2.31E10 571.5 mAb15160 318.2 10.2 55.8 3.04E+05 3.10E05 1.02E10 372.6 mAb15161 272 25.5 127 2.80E+05 1.38E05 4.90E11 840.0 mAb15162 303.6 18 115.1 1.05E+05 6.79E05 6.48E10 170.0 mAb15163 373.8 15.3 97.6 2.14E+05 1.15E05 5.62E11 1,002.6 mAb15164 331.2 15.7 107.4 2.98E+05 2.58E05 8.65E11 448.0 mAb15165 286.2 19.5 103.9 3.91E+05 1.09E05 2.79E11 1,058.7 mAb15166 264.4 15.1 87 2.76E+05 1.74E04 6.30E10 66.3 mAb15167 348.2 20.2 114.1 3.36E+05 1.59E04 4.72E10 72.8 mAb15170 290 10.6 48.4 1.20E+05 1.29E05 1.10E10 893.3 mAb15150 290.2 16 87.6 3.17E+05 1.00E05 3.15E11 1,155.0 mAb15151 375.4 21.7 144.9 3.35E+05 6.26E05 1.87E10 184.5 mAb1932 806.9 10.9 2.6 NB NB NB NB NB: No binding (NB) was observed under the current experimental conditions. IC: Observed binding did not fit to the binding simulation model and no binding kinetic parameters were determined under the current experimental conditions.

TABLE-US-00012 TABLE 12 Kinetic Binding Parameters of the Interaction of Anti-SARS-COV-2 Monoclonal Antibodies to Trimeric SARS-CoV2 Spike ECD foldon Fusion Protein at 37 C. mAb Spike Capture trimer.his Mab Level bound at k.sub.a k.sub.d KD t Captured (RU) 50 nM (1/Ms) (1/s) (M) (min) mAb14312 541.7 500 9.66E+05 5.44E05 5.63E11 212.4 mAb14260 519.4 495.3 9.49E+05 5.73E05 6.04E11 201.5 mAb14284 865.5 631.1 8.14E+05 2.05E05 2.51E11 564.8 mAb14258 518 523.9 1.02E+06 7.93E05 7.76E11 145.7 mAb14294 463.8 440.6 6.94E+05 2.29E05 3.30E11 503.9 mAb14283 535.5 470 8.65E+05 7.32E05 8.47E11 157.8 mAb14235 386.4 396.9 7.35E+05 3.80E05 5.17E11 304 mAb14289 463.7 455.7 9.11E+05 6.63E05 7.28E11 174.3 mAb13458 548.9 466 8.15E+05 4.08E05 5.00E11 283.2 mAb14313 687.8 610.7 1.01E+06 9.94E05 9.84E11 116.2 mAb14247 221.2 247 6.74E+05 4.72E05 7.01E11 244.6 mAb14315 576.2 457.2 7.43E+05 3.25E05 4.37E11 355.7 mAb13459 826.8 632.1 7.94E+05 4.76E05 5.99E11 242.9 mAb13457 468.9 452.4 7.67E+05 5.98E05 7.79E11 193.3 mAb14257 602.9 684.6 1.17E+06 1.65E04 1.41E10 70.1 mAb14255 516.8 575.4 1.03E+06 1.61E04 1.55E10 71.9 mAb14286 437.5 539.2 2.59E+06 1.41E04 5.44E11 82 mAb14282 497 566.5 1.05E+06 1.71E04 1.62E10 67.7 mAb14281 488.8 429.8 8.34E+05 1.16E04 1.39E10 99.7 mAb14230 372.3 435.6 9.06E+05 8.97E05 9.91E11 128.7 mAb14256 570.7 509.1 7.96E+05 2.42E04 3.04E10 47.7 mAb14285 533.8 647.3 1.16E+06 4.18E05 3.59E11 276.3 mAb14280 594.3 650.1 9.85E+05 2.86E05 2.90E11 403.7 mAb14234 564.5 473.8 6.99E+05 7.16E05 1.02E10 161.4 mAb14232 104.1 119.6 7.51E+05 1.33E04 1.77E10 87.1 mAb14292 504.7 236.7 5.92E+05 6.82E03 1.15E08 1.7 mAb14314 697.7 259.4 4.59E+05 6.48E03 1.41E08 1.8 mAb14248 505.9 2 NB NB NB NB mAb14295 558.8 6.1 NB NB NB NB mAb14233 558.6 1.5 NB NB NB NB mAb14287 523.6 5.2 NB NB NB NB mAb14293 537.9 3.5 NB NB NB NB mAb14291 511.8 1.1 NB NB NB NB mAb14249 478.6 2.7 NB NB NB NB mAb14316 306.5 4.8 NB NB NB NB mAb14296 660.5 2.5 NB NB NB NB mAb14231 489.4 2 NB NB NB NB mAb14288 571.6 2.9 NB NB NB NB mAb14297 530.7 2.5 NB NB NB NB mAb14290 457.4 4.3 NB NB NB NB mAb14259 395.1 3 NB NB NB NB mAb15156 463.7 4.1 474.1 1.89E+06 2.12E04 1.12E10 54.5 mAb15157 307.2 1.0 298.2 1.70E+06 2.40E04 1.41E10 48.2 mAb15158 295.7 1.3 224.5 7.58E+05 4.07E05 5.41E11 283.6 mAb15159 293.3 2.0 196.2 4.67E+05 3.22E05 6.86E11 359.1 mAb15160 400.3 1.4 301.9 1.26E+06 1.67E04 1.32E10 69.2 mAb15161 283.7 0.9 281.6 1.48E+06 9.14E05 6.16E11 126.4 mAb15162 323.0 1.5 205.5 4.40E+05 1.87E04 4.25E10 61.7 mAb15163 343.6 1.3 323.9 1.35E+06 4.85E05 3.64E11 238.1 mAb15164 387.4 1.5 379.2 1.78E+06 1.38E04 7.77E11 84.0 mAb15165 279.2 1.5 318.7 2.12E+06 1.15E04 5.42E11 100.2 mAb15166 265.2 0.5 282.8 1.54E+06 8.32E05 5.40E11 138.8 mAb15167 406.6 1.0 446.1 1.76E+06 9.78E05 5.57E11 118.1 mAb15170 303.3 1.4 221.3 8.46E+05 1.00E05 1.18E11 1,155.0 mAb15150 249.8 1.8 246.8 1.68E+06 3.42E05 2.02E11 337.6 mAb15151 595.2 0.8 372.7 1.92E+06 2.48E04 1.29E10 46.6 mAb1932 926.9 3.8 11.4 NB NB NB NB hBST: human B-cell Sorting Technology NB: No binding (NB) was observed under the current experimental conditions. IC: Observed binding did not fit to the binding simulation model and no binding kinetic parameters were determined under the current experimental conditions.

Example 4: Neutralization of SARS-CoV-2 Wild-Type and Variant Spike Proteins

[0261] To test whether anti-SARS-CoV-2 spike protein antibodies can neutralize SARS-CoV-2 variants, these antibodies were screened against a panel of VSV pseudotype viruses expressing wild-type and variant spike proteins.

Generation of Recombinant VSV

[0262] Non-replicative pseudoparticles were generated using a VSV genome encoding the firefly luciferase and GFP genes instead of the native viral glycoprotein (VSV-G). Infectious particles complemented with VSV-G (VSV-AG-Fluc-2A-GFP/VSV-G) were recovered and produced using standard techniques with minor modifications. HEK293T cells (ATCC CRL-3216) were plated on poly-lysine treated plates and incubated overnight in DMEM without glutamine (Life Technologies), 10% fetal bovine serum (Life Technologies) and 1% Penicillin/Streptomycin/L-glutamine (Life Technologies). The following day, the cells were transfected with the VSV genomic clone driven by a T7 promoter and helper plasmids expressing the VSV-N, VSV-P, VSV-G, VSV-L, and T7 RNA polymerase with Lipofectamine LTX reagent (Life Technologies). After 48 hours, the transfected cells were co-cultured with BHK-21 cells (ATCC CCL-10) transfected with VSV-G using the SE cell Line 4D-Nucleofector X Kit L (Lonza) in DMEM without glutamine (Life Technologies), 3% fetal bovine serum (Life Technologies) and 1% Penicillin/Streptomycin/L-glutamine (Life Technologies). Cells were monitored for GFP expression or cytopathic effect (CPE) indicative of virus replication. Virus was then plaque purified, expanded, and titered in BHK-21 cells transiently expressing VSV-G. Fully replicative VSV-SARS-CoV-2-S virus was generated by replacing the VSV glycoprotein with the native SARS-CoV-2 sequences encoding residues 1-1255 of the spike protein (NCBI Accession No. MN908947.3). VSV-SARS-CoV-2-Spike virus was recovered as described above but the HEK293T cells were instead cocultured with BHK-21 cells transfected with both VSV-G and hACE2. VSV-SARS-CoV-2-S virus was plaque-purified and titered in Vero cells (ATCC CCL-81) and expanded in Vero E6 cells (ATCC CRL-1586). After collection, stocks of both viruses were centrifuged at 3000g for 5 minutes to clarify, sucrose cushioned to concentrate 10-fold, aliquoted, and frozen at 80 C.

Pseudotyping of VSV

[0263] Non-replicative pseudoparticles were generated as previously described (Baum et al., Science 2020). Human codon-optimized SARS-CoV-2 spike (NCBI Accession No. MN908947.3) was cloned into an expression plasmid. A total of 1.2107 HEK293T cells (ATCC CRL-3216) were seeded overnight in 15-cm dishes in DMEM without glutamine (Life Technologies) containing 10% heat-inactivated fetal bovine serum (Life Technologies), and Penicillin-Streptomycin-L-Glutamine (Life Technologies). The following day, the cells were transfected with 15 g spike expression plasmid with Lipofectamine LTX (Life Technologies) following the manufacturer's protocol. At 24 hours post transfection, the cells were washed with phosphate buffered saline (PBS) and infected at a MOI of 1 with the VSV-G-Fluc-2A-GFP/VSV-G virus diluted in 10 mL Opti-MEM (Life Technologies). The cells were incubated 1 hour at 37C with 5% CO2. Cells were washed three times with PBS to remove residual input virus and overlaid with DMEM with glutamine (Life Technologies) with 0.7% IgG-free BSA (Sigma), sodium pyruvate (Life Technologies), and Gentamicin (Life Technologies). After 24 hours at 37 C. with 5% CO2, the supernatant containing pseudoparticles was collected, centrifuged at 3000g for 5 minutes to clarify, aliquoted, and frozen at 80 C. Variants were cloned into the spike expression plasmid using site-directed mutagenesis and pseudoparticles were produced as described above.

Neutralization Assays with VSV Based Pseudoparticles.

[0264] Vero cells (ATCC: CCL-81) were seeded in 96-well black, clear bottom tissue culture treated plated (Corning: 3904) at 20,000 cells/well in DMEM media without glutamine (Life Technologies) containing 10% heat-inactivated fetal bovine serum (Life Technologies), and 1 Penicillin/Streptomycin/L-Glutamine (Life Technologies) 24 hours prior to assay. Cells were allowed to reach approximately 85% confluence before use in assay. Antibodies were diluted in Infection Media containing DMEM with glutamine (Life Technologies), 0.7% Low IgG BSA (Sigma), 1 Sodium Pyruvate (Life Technologies), and 0.5% Gentamicin (Life Technologies) to 2 assay concentration and diluted 3-fold down in Infection media, for an 11-point dilution curve in the assay beginning at 3 g/mL (20 nM). Antibody dilutions were mixed 1:1 with pseudoparticles for 30 minutes at room temperature prior to addition onto Vero cells. Cells were incubated at 37 C., 5% CO2 for 24 hours. Supernatant was removed from cells prior to lysis with 100 L Glo Lysis Buffer (Promega). 100 L resuspended Bright Glo substrate (Promega) was then added and luminescence was read on a Spectramax i3x (Molecular Devices). Exported values were analyzed using GraphPad Prism (v8.4.1).

[0265] Individual monoclonal antibody half maximal inhibitory concentration (IC50) against VSV-SARS-CoV-2 spike protein (S)-expressing pseudovirus encoding the Wuhan-Hu-1 (NCBI Accession Number MN908947.3) sequence of spike protein (S-wt) or the D614G spike protein variant (SEQ ID NO: 1071) were determined in Vero cells (Table 13). The majority of antibodies displayed neutralization potency in the picomolar range (pM), with some exhibiting neutralization potency in nanomolar (nM) range, and some non-neutralizing. In addition, IC50s were measured for individual monolonal antibodies tested against certain variant of concern/interest VSV-SARS-CoV-2 spike protein (S)-expressing pseudoviruses (Table 14), including the Omicron variant (individual mutations contained in Omicron in Table 15; complete set of Omicron mutations in Table 16, and comparison to and combination with mAb10933 and mAb10987 in Table 17). In the Omicron neutralization assay, mAb15160 was unaffected, while mAb14315 had a marginally decreased neutralization. Based on these results, a mAb14315/mAb15160 combination and a mAb15160/mAb14256 combination maintain effective neutralization of the Omicron variant. In addition, mAb14284, mAb14235, and mAb14287 were potent neutralizers and were unaffected by the Omicron variants, while mAb15151 had a half-log decrease in neutralization of the Omicron variant but maintained potency nonetheless based on its strong neutralization properties. As shown in the tables, the lower the fold change from wt or D614G, the less impact the Omicron mutations have on neutralization. Additional mAb15160 and mAb14284 neutralization data is presented in Tables 18 and 19, showing that these antibodies are potent neutralizers of different variants, including Omicron lineages.

TABLE-US-00013 TABLE 13 mAb neutralization potency (IC50 (M)) against the wild-type or D614G strain of VSV-SARS-CoV-2-S pseudoparticles in Vero cells mAb IC50 (M) mAb13457 2.48E11 mAb13458 2.61E11 mAb13459 3.66E11 mAb14230 2.41E11 mAb14231 1.02E09 mAb14232 6.35E11 mAb14233 5.42E11 mAb14234 3.98E11 mAb14235 3.38E11 mAb14247 5.80E11 mAb14248 6.20E11 mAb14249 1.62E10 mAb14255 3.43E11 mAb14256 5.47E11 mAb14257 2.91E11 mAb14258 5.57E11 mAb14259 3.72E11 mAb14260 9.28E11 mAb14280 2.86E11 mAb14281 3.35E11 mAb14282 3.27E11 mAb14283 5.49E11 mAb14284 3.14E11 mAb14285 3.25E11 mAb14286 ~2.616E11.sup. mAb14287 1.99E10 mAb14288 1.38E09 mAb14289 4.02E11 mAb14290 1.20E09 mAb14291 1.23E10 mAb14292 5.77E11 mAb14293 1.10E09 mAb14294 5.67E11 mAb14295 1.95E10 mAb14296 5.26E10 mAb14297 3.85E10 mAb14312 4.66E11 mAb14313 2.72E10 mAb14314 no activity mAb14315 4.76E11 mAb14316 1.29E10 mAb15156 2.09E11 mAb15157 3.72E11 mAb15158 1.24E10 mAb15159 5.78E11 mAb15160 4.58E11 mAb15161 4.91E11 mAb15162 1.11E10 mAb15163 6.28E11 mAb15164 2.48E11 mAb15165 2.96E11 mAb15166 1.61E11 mAb15167 2.95E11 mAb15170 1.22E10 mAb15150 6.21E11 mAb15151 7.09E12 Underlined IC50 values determined using D614G spike protein.

TABLE-US-00014 TABLE 14 Pseudoparticle neutralization IC50 (M) of variants of concern/interest (VOCs/VOIs) variants in Vero cells B.1.1.7 B.1.351 P.1 E484K L452R mAb IC50 FC IC50 FC IC50 FC IC50 FC IC50 FC mAb14255 1.8E11 1.2 3.9E11 1.9 4.1E11 1.7 4.5E11 2.8 2.9E11 1.4 mAb14256 4.3E11 1.0 1.5E11 0.4 1.3E11 0.3 1.6E11 0.8 2.8E11 0.7 mAb14257 1.1E11 1.6 1.7E08 >48988 ND ND NA >2640 ND ND mAb14258 1.0E10 1.5 NA >1754 ND ND NA >794 ND ND mAb14312 ND ND 2.9E10 9.3 ND ND ND ND 7E11 2.2 mAb14315 8.0E11 2.7 9.8E12 0.3 4.1E11 0.6 1.5E11 0.5 4.2E11 1.0 mAb15156 3.1E11 1.5 4.8E12 0.2 ND ND ND ND 3.7E11 1.4 mAb15157 5.7E11 1.5 1.4E11 0.4 ND ND ND ND ND ND mAb15158 1.1E10 0.9 2.1E08 172 ND ND ND ND ND ND mAb15159 7.7E11 1.3 2.6E11 0.5 ND ND ND ND ND ND mAb15160 5.3E11 1.2 1.1E11 0.2 ND ND ND ND 5.8E11 1.6 mAb15161 7.2E11 1.5 1.5E11 0.3 ND ND ND ND 5.9E11 1.1 mAb15162 7.5E11 0.7 4.2E11 0.4 ND ND ND ND ND ND mAb15163 6.5E11 1.0 2.5E10 4.0 ND ND ND ND ND ND mAb15164 2.7E11 1.1 2.6E11 1.0 ND ND ND ND 3.0E11 2.0 mAb15165 2.9E11 1.0 1.6E11 0.6 ND ND ND ND 2.9E11 2.2 mAb15166 2.4E11 1.5 1.9E10 12.0 ND ND ND ND ND ND mAb15167 2.9E11 1.0 3.1E10 10.4 ND ND ND ND ND ND mAb15170 9.5E11 0.8 8.9E11 0.7 ND ND ND ND ND ND mAb15150 8.3E11 1.3 5.7E12 0.1 ND ND ND ND ND ND mAb15151 1.1E11 1.0 5.4E12 0.8 ND ND 2.3E11 2.1 2E11 0.9 Full B.1.1.7 (H69del, V70del, Y145del, N501Y, A570D, D614G, P681H, T716I, S982A, D1118H), B.1.351 (D80Y, D215Y, L241del, L242del, A243del, L242del, K417N, E484K, N501Y, D614G, A701V), and P.1 (L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I, V1176F) variants were assessed. Key RBD residues from B.1.429 (L452R) and B.1.526 (E484K) lineages were assessed. Fold-decrease in potency (FC) was calculated relative to wt or D614G control from same assay NA: No Activity ND: Not determined

TABLE-US-00015 TABLE 15 Pseudoparticle neutralization IC50 (M) and fold-change of variants in Vero cells Spike protein mAb14256 mAb14315 mAb15160 mAb10985 WT/D614G 2.606E11 2.95E11 4.58E11 2.17E10 (IC50) K417N 1.46 0.66 0.86 1.38 (FC) N440K 2.51 0.94 ND 1.41 (FC) S477N 1.01 0.60 1.01 1.32 (FC) T478K 0.81 0.96 1.98 0.98 (FC) Q493R 0.86 0.45 ND 0.38 (FC) N501Y 1.06 ND 1.81 1.28 (FC) Italics: IC50 values for each antibody against wt or D614G control spike protein Fold-decrease in potency (FC) was calculated relative to wt or D614G control from the same assay ND: Not determined

TABLE-US-00016 TABLE 16 Pseudoparticle neutralization IC50 (M), IC90 (M), and IC50 fold-change of Omicron variant SARS-CoV-2 in Vero cells SARS-CoV-2 spike mAb15160 + mAb14256 + glycoprotein mAb14256 mAb14315 mAb15160 mAb14315 mAb15160 D614G (control) IC50 5.33E11 4.97E11 2.95E11 4.76E11 4.84E11 (IC90, where provided) (4.08E10) Omicron IC50 NC (NC) 1.45E10 1.71E11 6.47E11 1.02E10 (IC90, where provided) D614G (control) IC50 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) FC (log10 FC) Omicron IC50 FC 375.38 (2.57) 2.91 (0.46) 0.58 (0.24) 1.36 (0.13) 2.10 (0.32) (log10 FC) SARS-CoV-2 spike glycoprotein mAb14284 mAb14235 mAb14294 mAb14234 mAb14312 D614G (control) 8.25E12 3.79E12 5.00E12 2.38E12 1.00E12 Omicron 1.03E11 6.84E12 6.67E12 2.62E12 7.94E13 D614G (control) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) IC50 FC (log10 FC) Omicron IC50 FC 1.25 (0.00) 1.81 (0.26) 1418.80 (3.15) 21.82 (1.34) 5287.59 (3.72) (log10 FC) SARS-CoV-2 spike glycoprotein mAb15170 mAb15162 mAb15150 mAb15151 mAb15159 mAb15161 D614G (control) 6.41E12 4.31E12 8.52E13 7.78E13 4.34E12 1.92E12 Omicron 6.11E12 7.25E12 1.23E12 2.44E12 5.05E12 1.08E12 D614G (control) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) IC50 FC (log10 FC) Omicron IC50 FC 37.20 (1.57) 27.88 (1.45) 392.47 (2.59) 3.14 (0.50) 214.77 (2.33) 160.43 (2.21) (log10 FC) SARS-CoV-2 spike glycoprotein mAb15157 mAb15156 mAb14287 mAb14297 D614G (control) 1.01E12 4.62E13 2.18E12 9.72E12 Omicron 1.15E12 4.96E13 4.42E12 NC D614G (control) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) 1.00 (0.00) IC50 FC (log10 FC) Omicron IC50 FC 19841.27 (4.30) 74.72 (1.87) 0.20 (0.69) 2058.67 (3.31) (log10 FC) Omicron spike protein used comprises SEQ ID NO: 1073 the following mutations: A67V, 69-70, T95I, G142D/143-145, 211/L212I, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, L981F NC: No IC50 calculated due to no activity FC: Fold change calculated relative to D614G control Values in bold and underlined: Fold change is at least this value, calculated as highest assay concentration/D614G IC50; highest concentration in assay was 2.00E08

TABLE-US-00017 TABLE 17 Pseudoparticle neutralization IC50 (M) and IC50 fold-change of Omicron variant (B.1.1.529/BA.1) SARS-CoV-2 in Vero cells mAb15160 + mAb10933 + mAb10933 + mAb10987 2:1:1 mAb10987 mAb15160 (inferred values) Fold Decrease Fold Decrease Fold Decrease in IC.sub.50 over in IC.sub.50 over in IC.sub.50 over Variant.sup.a IC.sub.50(M) Ref Virus.sup.b IC.sub.50(M) Ref Virus.sup.b IC.sub.50(M) Ref Virus.sup.b D614G control 9.91E12 1.0 1.92E11 1.0 3.84E11 1.0 B.1.1.529/BA.1 NC >2018.98.sup.c 3.18E11 1.65 6.36E11 3.3 (Omicron) .sup.aAmino acid substitutions versus Wuhan lineage S protein: B.1.1.529/BA.1 (A67V, del69-70, T95I, G142D/del143-145, del211/L212I, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, L981F) bFold decrease relative to reference (ref) D614G virus was calculated by dividing the IC.sub.50 value generated by the antibody(ies) in the presence of a particular variant by the IC.sub.50 value generated by the antibody(ies) in the presence of reference virus from the same assay. .sup.cWhere an IC.sub.50 value could not be accurately calculated, the change in mAb potency in the presence of each mutant was calculated by dividing the maximum antibody concentration tested (20 nM) by the IC.sub.50 value calculated for the reference D614G VLP. The actual fold change is at least that of the indicated value. Abbreviations: NC, not calculated due to poor or lack of neutralization

TABLE-US-00018 TABLE 18 IC.sub.50 and IC.sub.90 Values for mAb15160-Mediated Neutralization of Entry of pVSV-Luc- SARS-CoV-2-S Pseudotyped with S Protein Variants into Vero Cells mAb15160 mAb10933 (Casirivimab) mAb10987 (Imdevimab) Casirivimab + Imdevimab Fold Fold Fold Fold Change Change Change Change in IC.sub.50 in IC.sub.50 in IC.sub.50 in IC.sub.50 SARS-CoV-2 over over over over S Protein IC.sub.50 Ref IC.sub.50 IC.sub.90 Ref IC.sub.50 IC.sub.90 Ref IC.sub.50 IC.sub.90 Ref Variant [M] IC.sub.90 [M] Virus [M] [M] Virus [M] [M] Virus [M] [M] Virus Full Sequences or Key Residues of Variants under Surveillance.sup.a and Other Multi-Variant Lineages B.1.1.7 5.28E 2.21E10 1.15 2.26E 2.23E 0.69 1.71E11 9.31E 0.41 1.11E 9.09E 0.46 (Alpha) 11 11 10 11 11 11 B.1.351 (Beta) 1.07E 1.70E10 0.23 2.87E 2.68E 87.97 2.12E11 4.43E 0.51 2.67E 2.54E 1.11 11 09 08 11 11 10 P.1 (Gamma) 1.02E 6.55E11 0.48 7.52E 3.16E 167.54 4.58E12 6.46E 0.18 1.01E 7.63E 1.16 11 09 07 11 11 11 B.1.617.2 2.80E 1.55E10 3.07 4.32E 4.23E 0.55 1.04E11 2.11E 1.26 7.45E 5.02E 1.06 (Delta) 11 12 11 10 12 11 K417N + 5.61E 1.04E10 0.16 1.81E 5.01E 1.23 8.08E12 1.46E 0.58 7.50E 5.53E 0.55 L452R + 12 11 10 10 12 11 T478K (AY.1 [Delta]) L145H + 2.32E 1.82E10 1.73 1.15E 8.27E 0.70 3.60E11 6.57E 1.94 1.55E 7.38E 1.51 A222V + 11 11 11 10 11 11 L452R + T478K (AY4.2 [Delta]) L452R 5.83E 2.24E10 1.55 5.60E 3.17E 0.76 7.83E11 7.26E 1.03 4.96E 1.64E 1.15 (B.1.427/B.1. 11 11 10 10 11 10 429 [Epsilon]) E484K .sup.(B.1.526 [Iota]) 5.97E 4.83E10 2.84 2.49E 3.53E 5.55 2.62E11 4.30E 1.01 3.20E 1.60E 3.67 11 10 09 10 11 10 B.1.617.1 2.98E 1.76E10 2.98 1.14E 1.06E 22.12 1.51E11 1.63E 1.68 2.45E 9.08E 3.07 (Kappa) 11 10 09 10 11 11 F490S + L452Q 2.64E 2.35E10 0.73 1.37E 1.73E 0.93 4.23E11 5.72E 3.05 1.53E 1.23E 1.12 (C.37 11 11 10 10 11 10 [Lambda]) R346K + 2.52E 2.95E10 1.71 2.10E 1.74E 19.22 9.03E12 1.00E 0.60 1.44E 1.54E 1.29 E484K + 11 10 09 10 11 10 N501Y (B.1.621 [Mu]) BA.1 1.84E 1.80E10 1.05 NC NC >1731.60.sup.b NC NC >754.43.sup.b NC NC >1012.66.sup.b (Omicron) 11 BA.1.1 2.26E 1.57E10 0.96 NC NC >1336.01.sup.b NC NC >1108.65.sup.b NC NC >1460.92.sup.b (Omicron) 11 BA.2 1.46E 1.53E10 2.08 6.80E 8.04E 461.82 2.19E09 1.99E 158.12 2.60E 1.82E 190.26 (Omicron) 11 09 08 08 09 08 BA.2.12.1 5.63E 4.67E10 2.22 NC NC >702.26 3.18E09 1.79E 137.46 4.77E 1.71E 275.13 (Omicron) 11 08 09 08 BA.3 6.06E 1.16E09 3.34 NC NC 1258.65 NC NC 899.69 NC NC 1456.66 (Omicron) 11 BA.4/BA.5 NC NC >701.48 NC NC 652.76 1.38E09 1.10E 54.10 3.67E 1.91E 200.81 (Omicron) 08 09 08 N439K + E484K 3.57E 4.00E10 3.92 1.22E 1.98E 15.58 NC NC >2417.79.sup.b 4.23E 2.99E 60.04 (AV.1) 11 10 09 10 09 N440K + E484K 3.25E 4.37E10 3.57 2.03E 2.41E 25.89 1.85E09 1.07E 223.53 2.76E 1.60E 39.20 (B.1.619, 11 10 09 07 10 09 B.1.619.1, B.1.625) C.1.2 3.75E 2.03E10 2.79 1.76E 1.22E 10.69 8.62E12 8.51E 0.46 1.57E 8.03E 1.53 11 10 09 11 11 11 All Single-Mutation Variants Y145H 1.88E 1.03E10 1.40 1.61E 7.83E 0.98 2.13E11 1.51E 1.15 1.46E 4.36E 1.43 11 11 11 10 11 11 W152C 1.24E 1.60E10 0.34 4.99E 1.37E 0.34 9.60E12 1.05E 0.69 8.86E 1.01E 0.65 11 12 10 10 12 10 V308L 2.86E 2.58E10 0.79 1.39E 1.46E 0.95 1.62E11 1.60E 1.17 1.23E 1.05E 0.90 11 11 10 10 11 10 G339D 5.47E 2.42E10 1.46 2.54E 2.26E 0.94 2.52E11 1.98E 1.00 2.22E 9.66E 1.43 11 11 10 10 11 11 E340A 2.16E 2.02E10 0.74 2.36E 1.10E 0.78 2.13E11 2.26E 0.59 1.48E 1.02E 1.22 11 11 10 10 11 10 E340K 2.54E 1.81E10 0.86 1.98E 1.95E 0.65 1.63E11 4.20E 0.45 1.13E 1.36E 0.93 11 11 10 10 11 10 R346K 3.95E 2.56E10 2.99 2.45E 1.48E 1.44 2.90E11 1.85E 2.51 2.10E 8.42E 2.39 11 11 10 10 11 11 R346S 3.21E 1.65E10 1.36 2.31E 1.34E 1.54 2.76E11 2.28E 1.53 1.90E 8.63E 1.39 11 11 10 10 11 11 S371L 2.62E 1.01E09 7.00 1.37E 3.99E 5.07 1.78E10 6.48E 7.07 9.21E 2.52E 5.96 10 10 10 10 11 10 A372T 3.00E 1.57E10 0.16 2.01E 7.46E 0.05 4.09E12 8.55E 0.35 3.46E 4.35E 0.35 12 13 12 11 12 11 S373P 3.58E 1.66E10 0.96 1.90E 1.07E 0.71 1.88E11 1.16E 0.74 1.26E 7.11E 0.82 11 11 10 10 11 11 S375F 1.72E 7.65E11 0.46 8.68E 1.13E 0.32 7.45E12 2.75E 0.30 5.17E 4.04E 0.33 11 12 10 11 12 11 D389Y 6.90E 6.82E11 0.17 2.24E 3.73E 0.12 5.74E12 8.36E 0.41 4.69E 3.96E 0.24 12 12 11 11 12 11 D405N 3.26E 2.10E10 1.38 3.06E 2.69E 2.04 1.29E11 1.78E 0.71 1.73E 7.98E 1.27 11 11 10 10 11 11 E406D 1.91E 2.51E10 1.91 5.69E 5.05E 110.30 2.08E11 4.32E 2.31 3.67E 4.50E 4.60 11 10 09 10 11 10 E406Q 2.05E 1.72E10 2.07 1.07E 6.74E 255.89 1.12E11 1.63E 2.32 2.10E 2.99E 11.37 11 09 09 10 11 10 K417N 2.47E 2.29E10 1.27 8.96E 1.15E 8.74 6.22E12 7.65E 1.35 1.35E 5.16E 1.54 11 11 09 11 11 11 K417R.sup.c 5.04E 1.96E11 2.37 4.10E 2.96E 61.23 2.92E11 2.92E 1.70 1.24E 6.59E 2.45 12 09 08 10 10 10 N439K 2.13E 1.06E10 0.57 4.47E 4.31E 0.60 1.77E08 1.12E 231.51 7.25E 2.50E 1.69 11 11 10 07 11 10 N440D 5.41E 3.94E10 2.28 1.69E 2.09E 1.04 1.92E09 1.42E 119.13 6.71E 4.36E 4.90 11 11 10 08 11 10 N440K 2.44E 2.06E10 1.26 1.02E 1.22E 0.99 1.34E09 1.57E 291.40 1.88E 3.28E 2.15 11 11 10 08 11 10 V445T 2.23E 1.85E10 0.94 2.27E 1.82E 1.52 7.25E09 6. HE 401.72 4.86E 4.00E 3.55 11 11 10 OS 11 10 G446D 1.58E 2.80E10 0.44 1.75E 1.79E 1.19 7.66E09 3.38E 553.07 3.52E 2.71E 2.58 11 11 10 08 11 10 G446R 2.59E 1.72E10 1.09 1.09E 1.05E 0.68 4.41E09 4.26E 273.91 4.36E 2.74E 3.18 11 11 10 08 11 10 G446S 1.13E 1.74E10 0.58 8.70E 7.95E 0.85 4.24E09 1.61E 920.25 1.95E 2.24E 2.23 11 12 11 08 11 10 Y449F 2.03E 2.25E10 0.64 8.48E 6.34E 0.59 1.07E11 9.56E 0.96 6.23E 4.90E 0.82 11 12 11 11 12 11 Y449H 3.68E 2.12E10 1.26 2.74E 1.56E 0.91 3.11E12 6.68E 0.09 1.42E 1.43E 1.17 11 11 10 11 11 10 Y449N 2.68E 2.64E10 1.13 1.23E 1.16E 0.82 5.14E12 3.84E 0.28 7.83E 3.38E 0.57 11 11 10 11 12 11 N450K 4.75E 2.88E10 0.83 4.40E 1.97E 0.91 2.95E11 1.60E 0.84 3.18E 1.07E 0.91 11 11 10 10 11 10 L452R 5.83E 2.24E10 1.55 5.60E 3.17E 0.76 7.83E11 7.26E 1.03 4.96E 1.64E 1.15 11 11 10 10 11 10 L455F 3.15E 3.00E10 0.78 7.69E 9.21E 426.00 1.93E11 1.05E 1.39 2.73E 2.94E 1.39 11 09 08 10 11 10 K458N 4.50E 2.76E10 1.86 1.76E 1.89E 1.14 1.92E11 1.38E 1.60 1.39E 7.82E 2.58 11 11 10 10 11 11 K458R 2.20E 1.94E10 2.77 7.59E 1.09E 1.36 7.71E12 1.57E 0.96 6.58E 9.08E 1.28 11 12 10 10 12 11 A475V 1.47E 1.09E10 0.61 1.48E 1.49E 0.96 1.05E11 8.96E 0.88 8.43E 5.26E 1.56 11 11 10 11 12 11 S477N 3.78E 1.47E10 1.01 9.78E 4.22E 1.32 6.54E11 4.44E 0.86 4.15E 1.68E 0.97 11 11 10 10 11 10 S477R 3.87E 2.16E10 1.60 1.29E 1.14E 0.84 3.19E11 1.70E 2.67 1.19E 6.78E 2.19 11 11 10 10 11 11 T478I 4.19E 2.76E10 1.73 2.44E 1.84E 1.58 2.42E11 1.73E 2.02 1.58E 9.96E 2.93 11 11 10 10 11 11 T478K 2.45E 1.87E10 1.26 7.11E 9.04E 0.69 1.23E11 2.96E 2.66 6.28E 6.43E 0.72 11 12 11 10 12 11 P479S 2.63E 1.88E10 1.09 2.31E 1.76E 1.50 2.12E11 1.56E 1.77 1.03E 6.23E 1.90 11 11 10 10 11 11 E484A 5.09E 2.73E10 1.36 1.03E 7.12E 3.83 1.10E11 1.04E 0.43 1.87E 1.04E 1.21 11 10 10 10 11 10 E484K 5.97E 4.83E10 2.84 2.49E 3.53E 5.55 2.62E11 4.30E 1.01 3.20E 1.60E 3.67 11 10 09 10 11 10 G485S 2.30E 1.31E10 0.97 4.24E 3.46E 2.83 1.23E11 1.10E 0.68 1.93E 6.01E 1.41 11 11 10 10 11 11 F486V NC NC >531.21.sup.b NC NC >269.61.sup.b 5.75E11 2.40E 0.75 9.85E 3.68E 2.29 10 11 10 Y489H 2.53E 6.61E09 121.48 NC NC >2311.60.sup.b 3.55E12 7.16E 0.35 2.15E 2.42E 2.57 09 11 11 10 F490L 5.68E 2.08E10 1.94 2.96E 1.69E 0.98 2.37E11 1.51E 0.65 1.43E 1.53E 1.18 11 11 10 10 11 10 F490S 1.76E 2.69E10 0.60 2.53E 1.45E 0.84 7.24E12 4.51E 0.20 1.65E 1.81E 1.36 11 11 10 10 11 10 F490Y 1.86E 1.67E10 0.79 9.70E 1.47E 0.65 1.33E11 1.51E 0.74 1.62E 7.39E 1.18 11 12 10 10 11 11 Q493E 8.85E 6.10E11 1.11 NC NC >3588.73.sup.b 1.34E11 8.69E 1.67 3.05E 2.69E 5.95 12 11 11 10 Q493K 4.57E 6.68E10 3.40 6.25E 1.31E 378.67 1.77E11 1.53E 0.95 4.22E 3.40E 4.11 11 09 07 10 11 10 Q493R 9.47E 5.79E10 4.88 8.68E 8.94E 84.71 1.76E11 1.12E 3.82 2.79E 2.56E 3.18 11 10 09 10 11 10 G496S 4.53E 1.72E10 1.21 2.25E 1.07E 0.83 1.28E10 9.49E 5.08 2.66E 9.64E 1.72 11 11 10 10 11 11 Q498R 4.72E 2.55E10 1.26 1.89E 2.02E 0.70 1.93E11 2.00E 0.77 1.53E 7.85E 0.99 11 11 10 10 11 11 T500A 1.18E 6.09E10 0.12 3.74E 3.45E 0.90 4.91E11 4.77E 10.21 9.85E 1.05E 5.32 12 12 10 10 12 10 T500F 8.35E 1.97E10 0.84 2.76E 8.39E 0.66 1.70E12 2.56E 0.35 2.54E 1.90E 1.37 12 12 11 11 12 11 N501Y 5.40E 2.79E10 2.78 2.13E 2.08E 2.08 6.85E12 8.63E 1.49 1.03E 5.18E 1.17 11 11 10 11 11 11 Y505F 4.34E 3.08E10 1.07 2.06E 2.27E 1.14 1.84E11 1.50E 1.32 1.62E 1.10E 0.82 11 11 10 10 11 10 Y505H 2.84E 1.64E10 0.76 1.40E 8.52E 0.52 9.54E12 5.75E 0.38 1.40E 5.52E 0.91 11 11 11 11 11 11 .sup.aFull sequences and/or key residues of the S protein from variants under surveillance; full S protein sequences: B.1.1.7 (H69del, V70del, Y145del, N501Y, A570D, D614G, P681H, T716I, S982A, D1118H), B.1.351 (D80Y, D215G, L241del, L242del, A243del, K417N, E484K, N501Y, D614G, A701V), P.1 (L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I, V1176F), B.1.617.2 (T19R, G142D, E156G, F157del, R158del, L452R, T478K, D614G, P681R, D950N), B.1.617.1 (T95I, G142D, E154K, L452R, E484Q, D614G, P681R, Q1071H), BA.1 (A67V, del69-70, T95I, G142D/del143-145, del211/L212I, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, L981F), BA.1.1 (BA.1 + R346K), BA.2 (T19I, del24-26, A27S, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, S477N, T478K, E484A, Q493R, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K); key S protein residues: AY.1 (K417N + L452R + T478K), AY4.2 (Y145H + A222V + L452R + T478K), B.1.427/B.1.429 (L452R), B.1.526 (E484K), B.1.621 (R346K + E484K + N501Y), AV.1 (N439K + E484K), B.1.619/B.1.619.1/B.1.625 (N440K + E484K), C.1.2 (Y449H + E484K + N501Y). .sup.bWhere an IC.sub.50 value could not be accurately calculated, the change in mAb potency in the presence of each mutant was calculated by dividing the maximum antibody concentration tested (20 nM) by the IC.sub.50 value calculated for casirivimab, imdevimab, or casirivimab + timdevimab in the presence of reference pVSV-SARS-CoV-2-S pseudoparticles. .sup.cThe IC.sub.50 and IC.sub.90 values for neutralization of this variant by mAb15160 and imdevimab were compared to different reference virus datasets than casirivimab and casirivimab + imdevimab. del, deletion; ins, insertion; NA, not applicable; NC, not calculated due to poor or lack of neutralization; NT, not tested Fold change calculated relative to D614G control

TABLE-US-00019 TABLE 19 Summary of IC.sub.50 and IC.sub.90 Values for REGN14284-Mediated Neutralization of Entry of pVSV-Luc-SARS-CoV-2-S Pseudotyped with S Protein Variants into Vero Cells Fold Change in IC.sub.50 SARS-CoV-2 S IC.sub.50 IC.sub.90 over Ref Protein Variant [M] [M] Virus Full Sequences or Key Residues of Variants under Surveillance.sup.a and Other Multi-Variant Lineages Alpha B.1.1.7.sup.a 1.14E11 7.58E11 1.14 Beta B.1.351.sup.a 9.73E12 6.15E11 0.98 Gamma P.1.sup.a 8.34E12 7.52E11 0.84 Delta B.1.617.2.sup.a 6.09E12 6.07E11 0.69 K417N + L452R + T478K (AY.1) * 3.42E12 3.53E11 0.50 Y145H + A222V + L452R + T478K 2.53E11 2.34E10 0.75 (AY4.2) Epsilon W152C + L452R (B.1.427/B.1.429) 1.28E11 9.23E11 1.18 .sup.Iota E484K.sup.(B.1.526) 5.40E11 5.30E10 1.61 Kappa B.1.617.1.sup.a 4.24E11 3.84E10 1.26 Lambda F490S + L452Q (C.37) * 1.20E11 1.58E10 1.17 Mu R346K + E484K + N501Y (B.1.621) 9.89E12 8.66E11 0.99 Omicron BA.1.sup.a * 8.86E12 1.09E10 1.63 BA.1.1.sup.a 1.00E11 1.03E10 0.58 BA.2.sup.a * 1.13E11 1.54E10 0.98 BA.2.12.1.sup.a * 2.55E11 2.12E10 1.40 BA.3.sup.a 2.22E11 4.41E10 2.52 BA.4/BA.5.sup.a * 1.85E11 1.89E10 0.79 NA C.1.2 3.16E11 3.20E10 0.94 All Single-Mutation Variants W152C 9.40E12 6.46E11 0.81 S247R 3.70E12 3.53E11 0.90 V308L 1.38E11 1.30E10 1.19 N334D 6.69E12 1.75E10 1.01 G339D 4.01E11 2.86E10 1.68 E340A 1.43E11 1.28E10 1.86 E340K 1.16E11 8.07E11 1.51 R346K 1.04E11 9.43E11 1.41 R346S 2.73E11 1.05E10 1.59 S371F 4.66E11 2.80E10 12.75 S371Y 4.00E11 3.21E10 7.78 A372T 1.16E11 1.14E10 1.56 T376A 2.64E12 4.44E11 0.72 T376S 1.54E11 1.59E10 0.65 D405N 8.15E12 7.93E11 0.47 E406W 1.67E11 4.82E10 1.91 R408S 9.12E12 6.38E11 2.49 K417T 6.45E12 7.44E11 1.76 Y421F 4.47E12 8.04E11 0.51 N439K 4.13E11 8.23E10 1.98 N439V 1.59E11 1.97E10 0.76 N440D 5.16E11 5.34E10 2.47 N440K 3.09E11 7.31E10 1.48 L441F 1.80E11 1.36E10 0.86 L441Q 2.78E11 2.69E10 1.33 K444L NC NC >957.40.sup.b K444M 5.23E10 5.92E09 25.03 K444N NC NC >957.40.sup.b K444Q 4.93E09 5.90E08 236.00 K444R 4.33E12 4.92E11 1.54 K444T NC NC >5599.10.sup.b V445A 1.88E10 2.81E09 57.18 V445T 9.18E10 8.56E09 53.44 G446D 2.34E10 1.94E09 20.28 G446R 3.37E11 2.51E10 10.23 G446S 8.96E12 6.40E11 2.72 G446V 3.38E11 7.30E10 9.46 G447V 2.35E11 3.12E10 8.34 N448S 2.62E12 4.64E11 0.30 Y449D 8.48E12 1.01E10 2.57 Y449F 7.32E12 4.74E11 0.85 Y449H 1.04E11 8.45E11 1.36 Y449N 7.92E12 9.05E11 0.46 Y449S 8.70E12 8.60E11 2.64 N450K 8.36E12 1.19E10 0.72 L452M 8.87E12 6.13E11 2.16 L452Q 2.61E11 1.79E10 1.10 L452R 5.49E11 7.32E10 1.63 F456L 5.87E12 1.05E10 0.67 Y473F 6.01E12 8.57E11 0.68 A475V 7.47E12 8.37E11 0.65 S477N 1.37E11 2.02E10 1.70 T478K 1.34E11 1.69E10 1.66 E484A 1.38E11 1.74E10 0.58 E484K 5.40E11 5.30E10 1.61 E484Q 3.04E12 4.51E11 0.35 G485S 8.52E12 9.47E11 0.50 G485V 9.62E12 9.33E11 1.10 F486S 1.79E11 5.36E10 2.03 F486V 6.23E11 3.18E10 7.72 Y489H 8.93E13 4.43E11 0.15 F490L 2.12E11 1.41E10 2.76 F490S 2.04E11 1.19E10 2.65 F490Y 1.20E11 7.26E11 0.70 Q498H 1.10E11 1.41E10 3.09 Q498R 2.24E11 1.35E10 0.94 T500A 1.46E11 1.02E10 4.07 T500F 8.56E12 3.85E11 2.40 T500N 2.28E11 1.52E10 3.45 N501T 5.57E12 3.47E11 1.69 N501Y 1.22E11 1.42E10 1.51 Y505H 2.46E11 2.00E10 1.03 *The values shown for the indicated variant/reference virus represent the geometric mean from at least 3 replicate assays. .sup.aFull S protein sequences and/or key residues of variants under surveillance were assessed. Key residues assessed and the lineages they represent are shown in the table. Full sequences are indicated by footnote a within the table and contain the following substitutions in the Wuhan-Hu-1 S protein reference sequence: B.1.1.7 (H69del, V70del, Y145del, N501Y, A570D, D614G, P681H, T716I, S982A, D1118H), B.1.351 (D80Y, D215G, L241del, L242del, A243del, K417N, E484K, N501Y, D614G, A701V), P.1 (L18F, T20N, P26S, D138Y, R190S, K417T, E484K, N501Y, D614G, H655Y, T1027I, V1176F), B.1.617.2 (T19R, G142D, E156G, F157del, R158del, L452R, T478K, D614G, P681R, D950N), B.1.617.1 (T95I, G142D, E154K, L452R, E484Q, D614G, P681R, Q1071H), BA.1 (A67V, del69-70, T95I, G142D/del143-145, del211/L212I, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K, E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, L981F). BA.1.1 (BA.1 + R346K), BA.2 (T19I, del24-26, A27S, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, S477N, T478K, E484A, Q493R, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K), BA.2.12.1 (BA.2 + L452Q), BA.3 (G142D, G339D, S371F, S373P, S375F, D405N, K417N D614G, H655Y, N679K, P681H, D796Y, Q954H, N969K), BA.4/BA.5 (both lineages have an identical S protein sequence [T19I, L24del, P25del, P26del, A27S, H69del, V70del, G142D, V213G, G339D, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K, L452R, S477N, T478K, E484A, F486V, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K]) .sup.bWhere an IC.sub.50 value could not be accurately calculated, the change in mAb potency in the presence of each mutant was calculated by dividing the maximum antibody concentration tested (20 nM) by the IC.sub.50 value calculated for the antibody in the presence of reference pVSV-SARS-CoV-2-S pseudoparticles. del, deletion; ins, insertion; NC, not calculated due to poor or lack of neutralization mAb14284 was tested at concentrations ranging from approximately 300 fM to 20 nM. Fold change over reference (ref) virus was calculated by dividing the IC.sub.50 value determined for the antibody in the presence of a particular variant by the IC.sub.50 value determined for the antibody in the presence of reference pseudoparticles (pVSV-SARS-CoV-2 pseudotyped with the D614G mutation) from the same assay.

Example 5: Anti-SARS-CoV-2 Antibodies Block RBD Binding to hACE2 as Determined by ELISA

[0266] An ELISA-based blocking assay was used to determine the ability of anti-SARS-CoV-2 antibodies to block the binding of the SARS-COV-2 Spike protein receptor binding domain (RBD) to its receptor, human angiotensin converting enzyme 2 (hACE2).

[0267] The SARS-CoV-2 protein used in this assay was comprised of the receptor binding domain (RBD) portion of the SARS-CoV-2 Spike protein (amino acids Arg319-Phe541) expressed with a C-terminal myc-myc-6-Histidine tag (SARS-CoV-2 RBD.mmh). The human ACE-2 protein used in the experiments was purchased from R&D Systems, comprising a portion of the human ACE-2 extracellular domain (amino acids Leu18-Ser740) and a C-terminal 10-Histidine tag (hACE-2-10His; ACE2 NCBI Accession No. Q9BYF1).

[0268] Experiments were carried out using the following procedure. The hACE-2-10His receptor was coated at 2 g/ml in PBS on a 96-well microtiter plate overnight at 4 C. Nonspecific binding sites were subsequently blocked using a 0.5% (w/v) solution of BSA in PBS. In other microtiter plates, a constant amount of 300 pM of SARS-CoV-2 RBD.mmh protein was bound for one hour with anti-SARS-COV-2-S antibodies or an irrelevant human IgG1 antibody control at dilutions from 1.7 pM to 100 nM in PBS+0.5% BSA. The fixed concentration of SARS-CoV-2 RBD was selected to be near the concentration which generated 50% of the maximal binding (EC50 value) to the plate-adhered hACE-2. The antibody-protein complexes were transferred to the microtiter plate coated hACE-2-10His. After 1 hour of incubation at room temperature, the wells were washed, and plate-bound SARS-COV-2 RBD.mmh was detected with a polyclonal goat anti-myc antibody conjugated with horseradish peroxidase (HRP) (NovusBio). The plates were then developed using TMB substrate solution (BD Biosciences, #51-2606KC and #51-2607KC) according to the manufacturer's recommended procedure and absorbance at 450 nm was measured on a Victor X5 plate reader (PerkinElmer, Shelton CT).

[0269] Binding data were analyzed using a sigmoidal dose-response model within Prism software (GraphPad). The calculated IC50 value, defined as the concentration of antibody required to block 50% of SARS-CoV-2 RBD binding to plate-coated hACE2, was used as an indicator of blocking potency. Percent blocking of anti-SARS-CoV-2-S antibody at a given concentration was calculated based on the following formula:

[00002]$\%\mathrm{Blocking}=100-\left(\frac{\left[\mathrm{Experimental}{\mathrm{Signal}}_{\left(\mathrm{highest}\mathrm{Ab}\mathrm{conc}\right)}-\mathrm{Background}{\mathrm{Signal}}_{\left(\mathrm{buffer}\right)}\right]}{\left[\mathrm{Maximum}{\mathrm{Signal}}_{\left(\mathrm{hEGF}.\mathrm{mFc}\mathrm{alone}\right)}-\mathrm{Background}{\mathrm{Signal}}_{\left(\mathrm{buffer}\right)}\right]}100\right)$

[0270] Antibodies that blocked binding less than or equal to 50% at the highest concentration tested were classified as non-blockers and IC50 values were not reported for those antibodies.

[0271] The ability of anti-SARS-CoV-2 antibodies to block SARS-CoV-2 RBD binding to human ACE2 was assessed using a blocking ELISA. In this assay 300 pM SARS-COV-2 RBD.mmh was titrated with a wide concentration range of the anti-SARS-COV-2-S antibody. The inhibition of the binding of the SARS-COV-2 RBD to hACE-2 in the presence of the SARS-COV-2 antibody was evaluated. The plate-bound SARS-COV-2 RBD.mmh was detected with a HRP-conjugated goat anti-myc antibody. The IC50 values and maximum blocking at the highest tested concentrations of the SARS-COV-2 antibodies are summarized in Table 20.

[0272] Of the fifty-six antibodies tested, forty-two displayed antibody concentration-dependent blocking of SARS-COV-2 RBD binding to hACE-2 with maximum blocking ranging from 61% to about 100% at the highest antibody concentration tested (100 nM). The IC50 values for the identified blocking antibodies ranged from 102 pM to 19.7 nM. The remaining fourteen antibodies displayed less than 50% blocking activity at the highest concentration tested and were classified as non-blockers. An isotype control, mAb193281, was also used as a control.

TABLE-US-00020 TABLE 20 Blocking potency of Anti-SAR-COV-2-S Antibodies on Spike RBD-hFc Binding to Immobilized Human ACE-2 Antibody blocking 300 pM SARS-COV-2.mmh binding to hACE-2 % Blocking at 100 nM mAb mAb# IC.sub.50, M concentration mAb13457 1.97E08 75 mAb13458 8.73E10 81 mAb13459 4.43E09 78 mAb14230 9.97E09 85 mAb14231 Nbl 2 mAb14232 1.80E08 87 mAb14233 Nbl 7 mAb14234 4.62E09 92 mAb14235 2.34E10 96 mAb14247 4.46E10 95 mAb14248 Nbl 12 mAb14249 Nbl 12 mAb14255 1.57E10 92 mAb14256 4.80E10 95 mAb14257 3.65E10 95 mAb14258 1.21E10 93 mAb14259 1.03E08 89 mAb14260 1.27E10 96 mAb14280 1.74E09 98 mAb14281 3.97E10 100 mAb14282 5.02E10 96 mAb14283 3.73E10 97 mAb14284 3.67E10 98 mAb14285 1.70E09 96 mAb14286 6.41E10 94 mAb14287 Nbl 2 mAb14288 Nbl 5 mAb14290 Nbl 1 mAb14289 3.78E10 98 mAb14291 Nbl 8 mAb14292 2.54E10 95 mAb14293 Nbl 1 mAb14294 2.44E10 94 mAb14295 Nbl 5 mAb14296 Nbl 20 mAb14297 Nbl 16 mAb14312 1.34E10 98 mAb14313 1.64E08 63 mAb14314 Nbl 21 mAb14315 1.72E08 61 mAb14316 Nbl 9 mAb15156 2.69E10 103 mAb15157 3.57E10 103 mAb15158 3.09E10 103 mAb15159 1.56E10 102 mAb15160 2.04E10 96 mAb15161 5.84E10 94 mAb15162 7.56E10 95 mAb15163 2.17E10 95 mAb15164 2.45E10 99 mAb15165 1.89E10 99 mAb15166 4.75E10 98 mAb15167 1.06E09 96 mAb15170 1.02E10 82 mAb15150 2.54E10 98 mAb15151 6.41E10 98 mAb193281 Nbl 15 mAb193281 Nbl 16 Nbl: Non-Blocking-% blocking is less than or equal to 50%.

Example 6: PH Sensitivity of Anti-SARS-CoV-2-S Monoclonal Antibodies Binding to Monomeric SARS-CoV-2-S RBD Reagents Measured at 37 C.

[0273] The dissociation rate constants (k.sub.d) for 56 purified anti-SARS-CoV-2-S monoclonal antibodies at neutral and acidic pH conditions were determined using a real-time surface plasmon resonance (SPR)-based Biacore T200 or Biacore 4000 biosensor. All binding studies were performed at 37 C. using running buffers of 0.05% v/v Surfactant Tween-20-containing PBS at pH7.4, pH6.0, and pH5.0 (PBS-T-pH7.4, -pH6.0, and -pH5.0, respectively). The Biacore CM5 sensor chip surface was first derivatized by amine coupling with a mouse anti-human Fc specific mAb (Regeneron, mAb2567) to capture anti-SARS-CoV-2 monoclonal antibodies. The ligand examined for binding to anti-SARS-CoV-2 monoclonal antibodies was a recombinant protein comprising the SARS-COV-2 RBD extracellular domain expressed with a C-terminal tag of myc-myc-hexahistidine (SARS-COV-2 RBD-MMH; SEQ ID NO: 1069). Solutions of 90 nM SARS-COV-2 RBD-MMH were prepared in PBS-T-pH7.4 buffer and then injected over the antibody surfaces at a flow rate of 25 L/min for 3 minutes followed by a dissociation phase of bound SARS-COV-2 RBD-MMH with the running buffers at pH7.4, pH6.0, or pH5.0 flowing over for 8 minutes. An isotype control, mAb1932, was also used as a control.

[0274] The dissociation rate constants (k.sub.d) in three pH running buffers were determined by fitting the real-time binding sensorgrams to a 1:1 binding model using Scrubber 2.0c curve-fitting software. The dissociative half-life (t) was calculated from the k.sub.d values as:

[00003]$t^{1/2}\left(\min \right)=\frac{\ln \left(2\right)}{60*\mathrm{kd}}$

[0275] The resultant k.sub.d and t values for binding of monomeric SARS-COV-2 RBD-MMH to 56 anti-SARS-CoV-2 monoclonal antibodies captured on anti-hFc surfaces at pH6.0 and pH5.0 at 37 C. are summarized in Table 21 and Table 22, respectively, and the results at pH7.4 are listed in both tables for comparison.

TABLE-US-00021 TABLE 21 Comparison of the Dissociation Rate Constant and Dissociative Half-life of the Interaction of Monomeric SARS-COV-2 RBD-MMH to Anti-SARS-CoV-2 Monoclonal Antibodies at pH 7.4 and pH 6.0 at 37 C. mAb RBD.mmh mAb RBD.mmh t Ratio Capture Bound at kd at t Capture Bound at Kd at t at of mAb Level pH 7.4 pH 7.4 at pH 7.4 Level pH 7.4 pH 6.0 pH 6.0 pH 7.4 to Captured (RU) (RU) (1/s) (min) (RU) (RU) (1/s) (min) pH 6.0 mAb14284 386.5 146.9 6.24E04 18.5 369.4 144.5 5.74E04 20.1 0.9 mAb14312 276.1 95.2 8.59E04 13.5 257.6 94.8 1.77E03 6.5 2.1 mAb14294 222.4 83.2 9.20E04 12.6 207 91.1 5.65E04 20.5 0.6 mAb14315 238 83.3 9.80E04 11.8 217.5 82.5 9.46E04 12.2 1 mAb14292 241.2 73.4 1.41E03 8.2 218.6 74 1.78E03 6.5 1.3 mAb14235 202.9 71.2 1.48E03 7.8 185.2 80.8 1.14E03 10.1 0.8 mAb14247 137.5 40.8 1.58E03 7.3 107.4 43.8 1.67E03 6.9 1.1 mAb13458 290.7 96.5 1.61E03 7.2 265.5 98.6 1.99E03 5.8 1.2 mAb14283 246.6 87.7 1.69E03 6.9 224.5 86.3 2.00E03 5.8 1.2 mAb14289 246.7 86.1 1.85E03 6.2 224 84 2.72E03 4.2 1.5 mAb14260 208 74.5 1.88E03 6.2 192.7 83 1.66E03 6.9 0.9 mAb14258 245.1 85.9 2.13E03 5.4 221.1 85 5.48E04 21.1 0.3 mAb13457 309 103.3 2.14E03 5.4 279.3 100.8 3.09E03 3.7 1.5 mAb14234 133.8 37.9 2.21E03 5.2 103 44.5 1.43E03 8.1 0.6 mAb13459 301.6 106.8 2.29E03 5.1 290 114.8 2.39E03 4.8 1.1 mAb14313 275.7 94.2 2.51E03 4.6 255 97.1 4.10E03 2.8 1.6 mAb14255 288.4 97.5 4.93E03 2.3 262.3 96.4 5.13E03 2.3 1 mAb14281 256.9 83.5 6.59E03 1.8 233.5 85.2 7.34E03 1.6 1.1 mAb14230 224.5 67.6 7.71E03 1.5 193.5 70.7 7.97E03 1.4 1.1 mAb14257 234.8 82.3 7.78E03 1.5 220.2 91.3 8.42E03 1.4 1.1 mAb14286 259 84.3 9.68E03 1.2 234.5 87.2 9.99E03 1.2 1 mAb14256 258.4 80.2 1.11E02 1 234.3 84.4 9.55E03 1.2 0.8 mAb14282 215.3 73.5 1.26E02 0.9 199.9 83.5 9.85E03 1.2 0.8 mAb14280 255 72.4 2.40E02 0.5 230.3 78.4 1.90E02 0.6 0.8 mAb14285 213 64.2 2.58E02 0.4 200.2 77.1 1.44E02 0.8 0.5 mAb14232 58.1 8.2 3.11E02 0.4 34.4 23 1.24E02 0.9 0.4 mAb14259 233.1 12 IC IC 210.5 16.7 IC IC IC mAb14314 255.1 5.8 IC IC 244.4 12.5 IC IC IC mAb14231 272 0.1 NB NB 244.5 5 NB NB NB mAb14233 197 1.7 NB NB 164.3 3.3 NB NB NB mAb14248 230.5 0.9 NB NB 205.6 5.5 NB NB NB mAb14249 215.1 0.2 NB NB 198.1 8.7 NB NB NB mAb14287 254.4 1.1 NB NB 233 4.3 NB NB NB mAb14288 216.2 0.9 NB NB 201.3 8.8 NB NB NB mAb14290 245 0.3 NB NB 223.1 5.1 NB NB NB mAb14291 200.5 0.9 NB NB 185 9.5 NB NB NB mAb14293 257.1 1.4 NB NB 233.3 5.8 NB NB NB mAb14295 259.7 2.1 NB NB 239.1 1.6 NB NB NB mAb14296 276.5 1.7 NB NB 257.1 4.1 NB NB NB mAb14297 229 2.2 NB NB 214.7 9.3 NB NB NB mAb14316 201.7 2.1 NB NB 180.1 5.6 NB NB NB mAb15156 268.5 92.6 6.73E03 1.7 258.3 96.4 9.70E03 1.2 1.4 mAb15157 311.9 106.3 4.05E03 2.9 301.7 107.8 7.66E03 1.5 1.9 mAb15158 306.9 111.4 3.10E04 37 294.7 118.1 2.90E04 40.1 0.9 mAb15159 92.9 31.9 2.31E04 50 79.8 36.3 4.12E04 28.1 1.8 mAb15160 178.8 58.2 2.18E03 5.3 164.8 59.9 2.83E03 4.1 1.3 mAb15161 121.5 39.8 1.13E03 10.3 106.2 43.7 1.07E03 10.8 0.9 mAb15162 330.8 116.4 3.00E03 3.9 321.2 120.1 3.54E03 3.3 1.2 mAb15163 230.9 81.9 4.94E04 23.4 211.7 82.4 1.04E03 11.1 2.1 mAb15164 400.6 144.9 2.26E03 5.1 398.3 154.4 2.10E03 5.5 0.9 mAb15165 290.4 110 1.64E03 7.1 279.8 115 1.28E03 9.1 0.8 mAb15166 244.5 65.2 3.00E02 0.4 234.8 66 3.01E02 0.4 1 mAb15167 419.3 117.5 2.08E02 0.6 415.8 124.2 1.62E02 0.7 0.8 mAb15170 309.8 120.5 1.07E04 108.4 304.6 128.6 8.00E05 144.5 0.7 mAb15150 246.1 87.8 1.12E04 103.4 233.4 89.7 3.81E05 303.5 0.3 mAb15151 611.4 219.4 6.80E03 1.7 612.7 227.1 4.54E03 2.5 0.7 mAb1932 314 0.6 NB NB 294.3 7.7 NB NB NB NB: No binding (NB) was observed under the current experimental conditions. IC: Observed binding did not fit to the binding simulation model and no binding kinetic parameters were determined under the current experimental conditions.

TABLE-US-00022 TABLE 22 Comparison of the Dissociation Rate Constant and Dissociative Half-life of the Interaction of Monomeric SARS-COV-2 RBD-MMH (SEQ ID NO: 1069) to Anti-SARS- CoV-2 Monoclonal Antibodies at pH 7.4 and pH 5.0 at 37 C. mAb RBD.mmh mAb RBD.mmh t Ratio Capture Bound at kd at t at Capture Bound at Kd at t at of pH 7.4 mAb Level pH 7.4 pH 5.0 pH 5.0 Level pH 7.4 pH 5.0 pH 5.0 to Captured (RU) (RU) (1/s) (min) (RU) (RU) (1/s) (min) pH 5.0 mAb14284 386.5 146.9 6.24E04 18.5 387.2 158.8 7.36E04 15.7 1.2 mAb14312 276.1 95.2 8.59E04 13.5 274.5 96.7 5.58E03 2.1 6.4 mAb14294 222.4 83.2 9.20E04 12.6 210.4 97.6 4.52E04 25.5 0.5 mAb14315 238 83.3 9.80E04 11.8 228.3 101.2 7.00E04 16.5 0.7 mAb14292 241.2 73.4 1.41E03 8.2 229.2 85 2.19E03 5.3 1.5 mAb14235 202.9 71.2 1.48E03 7.8 184.9 80.8 1.19E03 9.7 0.8 mAb14247 137.5 40.8 1.58E03 7.3 107.1 51.9 1.91E03 6.1 1.2 mAb13458 290.7 96.5 1.61E03 7.2 270.3 90.9 4.01E03 2.9 2.5 mAb14283 246.6 87.7 1.69E03 6.9 237.7 91.7 5.12E03 2.3 3 mAb14289 246.7 86.1 1.85E03 6.2 235.5 89.3 4.60E03 2.5 2.5 mAb14260 208 74.5 1.88E03 6.2 193.8 82.4 2.02E03 5.7 1.1 mAb14258 245.1 85.9 2.13E03 5.4 230.8 103.4 5.08E04 22.7 0.2 mAb13457 309 103.3 2.14E03 5.4 293.7 104.7 4.52E03 2.6 2.1 mAb14234 133.8 37.9 2.21E03 5.2 98.5 40.4 1.66E03 7 0.7 mAb13459 301.6 106.8 2.29E03 5.1 293.9 105.8 3.70E03 3.1 1.6 mAb14313 275.7 94.2 2.51E03 4.6 265.2 80.8 5.73E03 2 2.3 mAb14255 288.4 97.5 4.93E03 2.3 274.8 94.1 6.35E03 1.8 1.3 mAb14281 256.9 83.5 6.59E03 1.8 241 51.7 1.17E02 1 1.8 mAb14230 224.5 67.6 7.71E03 1.5 201.1 60.3 9.24E03 1.3 1.2 mAb14257 234.8 82.3 7.78E03 1.5 224.7 45.2 1.72E02 0.7 2.1 mAb14286 259 84.3 9.68E03 1.2 246.3 82.4 6.64E03 1.7 0.7 mAb14256 258.4 80.2 1.11E02 1 239.7 54.1 9.27E03 1.2 0.8 mAb14282 215.3 73.5 1.26E02 0.9 201.9 62.4 6.71E03 1.7 0.5 mAb14280 255 72.4 2.40E02 0.5 241.4 61.7 1.03E02 1.1 0.5 mAb14285 213 64.2 2.58E02 0.4 202 51.5 2.50E03 4.6 0 mAb14232 58.1 8.2 3.11E02 0.4 22.3 10.1 6.56E04 17.6 0 mAb14259 233.1 12 IC IC 214.1 1.8 IC IC IC mAb14314 255.1 5.8 IC IC 252.9 3.4 IC IC IC mAb14231 272 0.1 NB NB 249.9 1.3 NB NB NB mAb14233 197 1.7 NB NB 173.5 6.6 NB NB NB mAb14248 230.5 0.9 NB NB 210.3 0.8 NB NB NB mAb14249 215.1 0.2 NB NB 198.6 1.9 NB NB NB mAb14287 254.4 1.1 NB NB 240.6 0.3 NB NB NB mAb14288 216.2 0.9 NB NB 203.7 4 NB NB NB mAb14290 245 0.3 NB NB 228.9 0.6 NB NB NB mAb14291 200.5 0.9 NB NB 188.5 3.2 NB NB NB mAb14293 257.1 1.4 NB NB 243.1 0.2 NB NB NB mAb14295 259.7 2.1 NB NB 250.3 8.7 NB NB NB mAb14296 276.5 1.7 NB NB 267.7 1 NB NB NB mAb14297 229 2.2 NB NB 218.2 3.1 NB NB NB mAb14316 201.7 2.1 NB NB 181.3 0.9 NB NB NB mAb15156 268.5 92.6 6.73E03 1.7 259.6 103.6 1.58E02 0.7 2.3 mAb15157 311.9 106.3 4.05E03 2.9 301.7 107.5 7.02E03 1.6 1.7 mAb15158 306.9 111.4 3.10E04 37 296.4 121.6 9.70E05 118.9 0.3 mAb15159 92.9 31.9 2.31E04 50 73.6 45.5 4.29E04 27 1.9 mAb15160 178.8 58.2 2.18E03 5.3 160.8 61.3 2.28E03 5.1 1 mAb15161 121.5 39.8 1.13E03 10.3 100.8 51.4 9.80E04 11.8 0.9 mAb15162 330.8 116.4 3.00E03 3.9 335.3 130.8 5.40E03 2.1 1.8 mAb15163 230.9 81.9 4.94E04 23.4 221.3 83.2 3.77E03 3.1 7.6 mAb15164 400.6 144.9 2.26E03 5.1 409.6 159 2.00E03 5.8 0.9 mAb15165 290.4 110 1.64E03 7.1 289.5 126.8 9.73E04 11.9 0.6 mAb15166 244.5 65.2 3.00E02 0.4 233.6 60 3.50E02 0.3 1.2 mAb15167 419.3 117.5 2.08E02 0.6 422.4 126 1.33E02 0.9 0.6 mAb15170 309.8 120.5 1.07E04 108.4 314.3 141.1 6.62E05 174.4 0.6 mAb15150 246.1 87.8 1.12E04 103.4 236.7 92.6 1.00E05 1155 0.1 mAb15151 611.4 219.4 6.80E03 1.7 632.9 230.7 4.39E03 2.6 0.6 mAb1932 314 0.6 NB NB 305.2 1.9 NB NB NB NB: No binding (NB) was observed under the current experimental conditions. IC: Observed binding did not fit to the binding simulation model and no binding kinetic parameters were determined under the current experimental conditions.

Example 7: Cross-Competition Between Anti-SARS-CoV-2-S Antibodies

[0276] Binding competition between 41 anti-SARS-CoV-2-S monoclonal antibodies (mAbs) was determined using a real time, label-free bio-layer interferometry (BLI) assay on the Octet HTX biosensor platform (Pall ForteBio Corp.). The entire experiment was performed at 25 C. in a buffer containing 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% v/v Surfactant Tween-20, and 1 mg/mL BSA, pH7.4 (HBS-EBT) with the sample plate shaking at a speed of 1000 rpm. To assess whether mAbs compete with one another for binding to their respective epitopes on SARS-COV-2 RBD extracellular domain expressed with a C-terminal myc-myc-hexahistidine (SARS-COV-2 RBD-MMH; SEQ ID NO: 1069) tag, 0.52-0.63 nm of SARS-COV-2 RBD-MMH was first captured onto anti-Penta-His antibody coated Octet biosensor tips (Fortebio Inc, #18-5122) by submerging the biosensor tips for 90 seconds in wells containing 10 g/mL solution of SARS-COV-2 RBD-MMH. The SARS-COV-2 RBD-MMH captured biosensor tips were then saturated with a first anti-SARS-CoV-2 monoclonal antibody (mAb-1) by dipping into wells containing 50 g/mL mAb-1 for 4 minutes. The tips were then submerged into wells containing 50 g/mL of a second anti-SARS-CoV-2 monoclonal antibody (mAb-2) for 4 minutes. Between steps the tips were rinsed with HBS-ETB. The real-time binding response was monitored during the entire course of the experiment and the binding response at the end of every step was recorded. The response of mAb-2 binding to SARS-COV-2 RBD-MMH pre-complexed with mAb-1 was compared to the mAb-2 binding signal where the mAb-1 was a nonbinding isotype control, and the percentage reduction of mAb-2 binding was calculated. If pre-binding of mAb-1 resulted in a >50% reduction of mAb-2 binding, and reversing the order of binding for the mAb pair also exhibited >50% reduction in binding, mAb-1 and mAb-2 were classified as competing mAbs. FIG. 1 displays the results from cross-competition studies for anti-SARS-CoV-2 mAbs in a matrix format. Table 23 lists the competing mAbs for each anti-SARS-CoV-2 mAb included in the experiment.

TABLE-US-00023 TABLE 23 Cross-competing anti-SARS-CoV-2-S monoclonal antibodies upon binding to immobilized SARS-COV-2 RBD-MMH (SEQ ID NO: 1069) mAb-1 mAb-2 Competing with mAb-1 mAb14312 mAb14256 mAb14255 mAb14257 mAb14285 mAb14286 mAb14280 mAb14282 mAb14281 mAb14256 mAb14312 mAb14286 mAb14280 mAb14282 mAb14284 mAb10985 mAb14255 mAb14312 mAb14258 mAb14257 mAb14285 mAb14286 mAb14280 mAb14282 mAb10933 mAb14258 mAb14257 mAb14285 mAb14286 mAb14280 mAb14282 mAb14258 mAb14255 mAb10933 mAb14258 mAb14257 mAb14285 mAb14286 mAb14280 mAb14282 mAb14283 mAb14257 mAb14312 mAb14255 mAb10933 mAb14258 mAb14286 mAb14280 mAb14282 mAb14313 mAb14283 mAb14285 mAb14312 mAb14255 mAb10933 mAb14258 mAb14286 mAb14280 mAb14282 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14286 mAb14312 mAb14256 mAb14255 mAb10933 mAb14258 mAb14257 mAb14285 mAb14280 mAb14282 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14260 mAb14280 mAb14312 mAb14256 mAb14255 mAb10933 mAb14258 mAb14257 mAb14285 mAb14286 mAb14282 mAb14281 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14282 mAb14312 mAb14256 mAb14255 mAb10933 mAb14258 mAb14257 mAb14285 mAb14286 mAb14280 mAb14281 mAb10987 mAb14289 mAb14281 mAb14312 mAb14280 mAb14282 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14315 mAb13457 mAb14285 mAb14286 mAb14280 mAb14281 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14294 mAb14235 mAb14234 mAb14247 mAb14260 mAb14315 mAb14230 mAb14284 mAb13458 mAb14285 mAb14286 mAb14280 mAb14281 mAb13457 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14294 mAb14235 mAb14234 mAb14247 mAb14260 mAb14315 mAb14230 mAb14284 mAb13459 mAb14285 mAb14286 mAb14280 mAb14281 mAb13457 mAb13458 mAb14313 mAb14283 mAb10987 mAb14289 mAb14294 mAb14235 mAb14234 mAb14247 mAb14260 mAb14315 mAb14230 mAb14284 mAb14313 mAb14257 mAb14285 mAb14286 mAb14280 mAb14281 mAb13457 mAb13458 mAb13459 mAb14283 mAb10987 mAb14289 mAb14294 mAb14235 mAb14234 mAb14247 mAb14260 mAb14315 mAb14230 mAb14284 mAb14283 mAb14258 mAb14257 mAb14285 mAb14286 mAb14280 mAb14281 mAb13457 mAb13458 mAb13459 mAb14313 mAb10987 mAb14289 mAb14294 mAb14235 mAb14234 mAb14247 mAb14260 mAb14315 mAb14230 mAb14284 mAb10987 mAb14285 mAb14286 mAb14280 mAb14282 mAb14281 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb14289 mAb14235 mAb14234 mAb14247 mAb14260 mAb14315 mAb14230 mAb14284 mAb14289 mAb14285 mAb14286 mAb14280 mAb14282 mAb14281 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14294 mAb14235 mAb14234 mAb14247 mAb14260 mAb14230 mAb14284 mAb14294 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb14289 mAb14235 mAb14234 mAb14247 mAb14230 mAb14284 mAb14235 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14294 mAb14234 mAb14247 mAb14260 mAb14315 mAb14230 mAb14284 mAb14234 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14294 mAb14235 mAb14247 mAb14260 mAb14315 mAb14230 mAb14284 mAb14247 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14294 mAb14235 mAb14234 mAb14260 mAb14315 mAb14230 mAb14284 mAb14260 mAb14286 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14235 mAb14234 mAb14247 mAb14315 mAb14230 mAb14284 mAb14315 mAb14281 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14235 mAb14234 mAb14247 mAb14260 mAb14230 mAb14230 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14294 mAb14235 mAb14234 mAb14247 mAb14260 mAb14315 mAb14284 mAb14284 mAb14256 mAb13457 mAb13458 mAb13459 mAb14313 mAb14283 mAb10987 mAb14289 mAb14294 mAb14235 mAb14234 mAb14247 mAb14260 mAb14230 mAb10985 mAb14256

[0277] Binding competition between 15 additional anti-SARS-Cov-2-S monoclonal antibodies was performed substantially as described above. Competition was determined using a real time, label-free bio-layer interferometry (BLI) assay on the Octet HTX biosensor platform (Pall ForteBio Corp.). The entire experiment was performed at 25 C. in a buffer containing 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% v/v Surfactant Tween-20, and 1 mg/mL BSA, pH7.4 (HBS-EBT) with the plate shaking at a speed of 1000 rpm. To assess whether mAbs are able to compete with one another for binding to their respective epitopes on SARS-COV-2 RBD extracellular domain expressed with a C-terminal myc-myc-hexahistidine (SARS-COV-2 RBD-MMH; SEQ ID NO: 1069) tag, 0.51 nm of SARS-COV-2 RBD-MMH was first captured onto anti-Penta-His antibody coated Octet biosensor tips (Fortebio Inc, #18-5122) by submerging the biosensor tips for 90 seconds in wells containing 10 g/mL of SARS-COV-2 RBD-MMH. The SARS-COV-2 RBD-MMH captured biosensor tips were then saturated with a first anti-SARS-CoV-2 monoclonal antibody (mAb-1) by dipping into wells containing 50 g/mL mAb-1 for 4 minutes. The biosensor tips were then submerged into wells containing 50 g/mL of a second anti-SARS-CoV-2 monoclonal antibody (mAb-2) for 4 minutes. Between steps the biosensor tips were rinsed with HBS-ETB buffer. The real-time binding response was monitored during the entire course of the experiment and the binding response at the end of every step was recorded. The response of mAb-2 binding to SARS-COV-2 RBD-MMH pre-complexed with mAb-1 was compared to the mAb-2 binding signal where the mAb-1 was a nonbinding isotype control, and the percentage reduction of mAb-2 binding was calculated. If pre-binding of mAb-1 resulted in >50% reduction of mAb-2 binding, and reversing the order of binding for the mAb pair also exhibited >50% reduction of binding, mAb-1 and mAb-2 were classified as competing mAbs. FIG. 2 displays the results from cross-competition studies for anti-SARS-Cov-2 mAbs in a matrix format. Table 24 lists the competing mAbs for each anti-SARS-CoV-2 mAb included in the experiment.

TABLE-US-00024 TABLE 24 Cross-competing anti-SARS-CoV-2-S monoclonal antibodies upon binding to immobilized SARS-COV-2 RBD-MMH (SEQ ID NO: 1069) mAb-1 mAb-2 Competing with mAb-1 mAb10934 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15164 mAb15165 mAb15161 mAb15160 mAb10933 mAb15163 mAb14312 mAb14232 mAb10987 mAb14289 mAb14283 mAb14247 mAb14234 mAb14230 mAb14315 mAb14284 mAb14235 mAb14294 mAb10989 mAb10934 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10987 mAb14289 mAb14283 mAb15158 mAb10934 mAb10989 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10985 mAb14289 mAb15156 mAb10934 mAb10989 mAb15158 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb14289 mAb14283 mAb15157 mAb10934 mAb10989 mAb15158 mAb15156 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10985 mAb14247 mAb15166 mAb10934 mAb10989 mAb15158 mAb15156 mAb15157 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb15167 mAb10989mAb mAb15158 mAb15156 mAb15157 mAb15166 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb15150 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb10954 mAb15162 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10985 mAb10954 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10985 mAb15162 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10985 mAb15159 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb10954 mAb15162 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10985 mAb15170 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10985 mAb15164 mAb10934 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb15165 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb15151 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb15161 mAb10934 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb15160 mAb10934 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb10933 mAb10934 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb15163 mAb14255 mAb14256 mAb14312 mAb10986 mAb15163 mAb10934 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb14255 mAb14256 mAb14312 mAb10986 mAb14255 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14256 mAb14312 mAb10986 mAb14256 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14312 mAb10986 mAb10985 mAb14312 mAb10934 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb10986 mAb10986 mAb10989 mAb15158 mAb15156 mAb15157 mAb15166 mAb15167 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb15164 mAb15165 mAb15151 mAb15161 mAb15160 mAb10933 mAb15163 mAb14255 mAb14256 mAb14312 mAb10985 mAb10985 mAb15158 mAb15157 mAb15150 mAb10954 mAb15162 mAb15159 mAb15170 mAb14256 mAb10986 mAb14232 mAb10934 mAb10987 mAb14289 mAb14283 mAb14247 mAb14234 mAb14230 mAb14315 mAb14284 mAb14235 mAb14294 mAb10987 mAb10934 mAb10989 mAb14232 mAb14289 mAb14283 mAb14247 mAb14234 mAb14230 mAb14315 mAb14284 mAb14235 mAb14294 mAb14289 mAb10934 mAb10989 mAb15158 mAb15156 mAb14232 mAb10987 mAb14283 mAb14247 mAb14234 mAb14230 mAb14315 mAb14284 mAb14235 mAb14294 mAb14283 mAb10934 mAb10989 mAb15158 mAb15156 mAb14312 mAb14232 mAb10987 mAb14289 mAb14247 mAb14234 mAb14230 mAb14315 mAb14284 mAb14235 mAb14294 mAb14247 mAb10934 mAb14232 mAb10987 mAb14289 mAb14283 mAb14234 mAb14230 mAb14315 mAb14284 mAb14235 mAb14294 mAb14234 mAb10934 mAb14232 mAb10987 mAb14289 mAb14283 mAb14247 mAb14230 mAb14315 mAb14284 mAb14235 mAb14294 mAb14230 mAb10934 mAb14232 mAb10987 mAb14289 mAb14283 mAb14247 mAb14234 mAb14315 mAb14284 mAb14235 mAb14294 mAb14315 mAb10934 mAb14232 mAb10987 mAb14289 mAb14283 mAb14247 mAb14234 mAb14230 mAb14284 mAb14235 mAb14294 mAb14284 mAb10934 mAb14232 mAb10987 mAb14289 mAb14283 mAb14247 mAb14234 mAb14230 mAb14315 mAb14235 mAb14294 mAb14235 mAb10934 mAb14232 mAb10987 mAb14289 mAb14283 mAb14247 mAb14234 mAb14230 mAb14315 mAb14284 mAb14294 mAb14294 mAb14232 mAb10987 mAb14289 mAb14283 mAb14247 mAb14234 mAb14230 mAb14315 mAb14284 mAb14235

Example 8: Characterization of Anti-SARS-CoV-2-S mAbs in an ADCC Surrogate Assay (FcR3a Val176 Signaling Assay)

[0278] The ability of antibodies targeting the spike protein of SARS-CoV-2 to interact with FcR3a, an Fc-receptor prominently expressed on NK cells that induces antibody dependent cell-mediated cytotoxicity (ADCC), was measured in a surrogate bioassay using reporter cells and target cells bound to antibodies. In this assay, engineered Jurkat T cells expressed the reporter gene luciferase under the control of the transcription factor NFAT (NFAT-Luc) along with the high affinity human FcR3a 176Val allotype receptor (Jurkat/NFAT-Luc/hFcR3a 176Val). Target cells were engineered Jurkat T cells expressing human CD20 (used as a positive control with CD20 targeting human IgG1 antibody) alone or in combination with the full-length SARS-CoV-2 spike protein. Reporter cells were incubated with target cells, and engagement of FcR3a via the Fc domain of human IgG1 antibodies bound to target cells led to the activation of the transcription factor NFAT in the reporter cells and drove the expression of luciferase which was then measured via a luminescence readout.

Target Cells

[0279] Jurkat/hCD20: Jurkat T cells were engineered to constitutively express full length human CD20 (amino acids M1-P297 of NCBI accession number NP_690605.1). Jurkat/hCD20 cells were stained for CD20 expression and maintained in RPMI+10% FBS+P/S/G+250 g/ml hygromycin growth medium. These cells were used as a negative control.

[0280] Jurkat/hCD20/SARS-CoV-2 spike: Jurkat/hCD20 T cells were engineered to constitutively express full-length SARS-CoV-2 spike protein (amino acids M1-T1273 of NCBI accession number YP_009724390.1). Jurkat/hCD20/SARS-CoV-2 spike cells were sorted for high expression of the spike protein and maintained in RPMI+10% FBS+P/S/G+1 g/ml puromycin+250 g/ml hygromycin growth medium.

Reporter Cells

[0281] Jurkat/NFAT-Luc/FcR3a 176Val: Jurkat T cells were engineered to stably express a Nuclear Factor of Activated T-cells (NFAT) luciferase reporter construct along with the high affinity human FcR3a 176Val allotype receptor (amino acids M1-K254 of accession number P08637 VAR_003960). Engineered reporter cells were maintained in RPMI1640+10% FBS+P/S/G+0.5 g/ml puromycin+500 g/ml G418 growth media.

Assay Set-Up

[0282] One day before the experiment, Jurkat reporter cells were split to 7.510.sup.5 cells/ml in RPMI1640+10% FBS+P/S/G+0.5 g/ml puromycin+500 g/ml G418 growth media. Jurkat target cells were split to 510.sup.5 cells/ml in RPMI1640+10% FBS+P/S/G+0.5 g/ml puromycin+250 g/ml hygromycin growth media.

[0283] On the day of the experiment, the target and reporter cells were transferred into assay media (RPMI+10% FBS+P/S/G) and added at a 3:2 ratio (310.sup.4/well target cells and 210.sup.4/well reporter cells) to 384-well white microtiter plates.

[0284] Anti-SARS-CoV-2 spike protein antibodies, a negative isotype-matched control antibody, and a positive control antibody for ADCC (anti-CD20) were titrated in an 11-point, 1:3 serial dilution ranging from 1.7 pM to 100 nM final concentration, with the final 12th point containing no antibody. All samples were tested in duplicates. Plates were incubated at 37 C./5% C02 for 5 h followed by the addition of an equal volume of ONE-Glo (Promega) reagent to lyse cells and detect luciferase activity. The emitted light was captured in Relative Light Units (RLU) on a multi-label plate reader Envision (PerkinElmer). EC50 values of the antibodies were determined from a 4 parameter logistic equation over a 12-point dose response curve (including the background signal) using GraphPad Prism software. Maximum fold induction was calculated using the following equation:

Fold Induction=Max Average RLU within tested dose range of each antibody/Average RLU (background signal=no antibody)

[0285] The EC50 values and fold inductions are summarized in Table 25.

Results with Jurkat/hCD20/SARS-CoV-2 Spike Target Cells

[0286] Each of the tested anti-SARS-CoV-2-S antibodies showed greater fold induction than the negative control antibody. The EC50 values ranged from 60.4 pM to 3.4 nM, and the fold induction values ranged from 3.9 to 13.5. The positive CD20 control antibody showed a fold induction of 29.9 with an EC50 value of 18.1 pM.

Results with Jurkat/hCD20 Target Cells

[0287] Each of the 41 anti-SARS-CoV-2-S antibodies except one (mAb14312) behaved similarly to the negative control antibody showing minimal activity. mAb14312 had an EC50 value of 18.4 nM and a fold induction value of 2.0. The positive CD20 control antibody showed a fold induction value of 29.4 with an EC50 value of 33.7 pM.

TABLE-US-00025 TABLE 25 Surrogate ADCC assay results Jurkat CD20 Jurkat/CD20/spike Fold Fold mAb # EC50 (M) induction EC50 (M) induction mAb13457 ND 1.2 1.50E10 9.1 mAb13458 ND 1.2 1.43E10 10.2 mAb13459 ND 1.2 1.42E10 9.9 mAb14230 ND 1.1 1.79E10 9.8 mAb14231 ND 1.2 3.25E09 7.9 mAb14232 ND 1.2 1.31E10 10.8 mAb14233 ND 1.1 2.80E10 11.0 mAb14234 ND 1.1 2.69E10 12.6 mAb14235 ND 1.0 1.43E10 10.8 mAb14247 ND 1.0 1.79E10 11.3 mAb14248 ND 1.0 1.79E10 12.8 mAb14249 ND 1.2 2.51E10 10.3 mAb14255 ND 1.0 1.67E10 9.5 mAb14256 ND 1.0 4.48E10 6.1 mAb14257 ND 1.0 2.39E10 8.8 mAb14258 ND 1.0 2.82E10 7.5 mAb14259 ND 1.0 3.94E10 8.7 mAb14260 ND 1.1 3.63E10 6.0 mAb14280 ND 1.1 2.67E10 13.5 mAb14281 ND 1.1 1.65E10 10.3 mAb14282 ND 1.1 2.57E10 10.7 mAb14283 ND 1.1 1.39E10 9.1 mAb14284 ND 1.1 1.56E10 9.0 mAb14285 ND 1.1 2.43E10 11.0 mAb14286 ND 1.0 2.04E10 11.0 mAb14287 ND 1.0 1.23E09 6.2 mAb14288 ND 1.0 5.10E10 6.7 mAb14289 ND 1.0 1.30E10 6.3 mAb14290 ND 1.0 3.42E09 9.5 mAb14291 ND 1.1 3.02E10 9.7 mAb14292 ND 1.0 3.86E10 9.2 mAb14293 ND 1.1 7.88E10 9.3 mAb14294 ND 1.0 6.04E11 10.5 mAb14295 ND 1.0 1.93E10 11.5 mAb14296 ND 1.0 4.11E10 8.1 mAb14297 ND 1.0 1.24E10 7.9 mAb14312 1.84E08 2.0 1.68E10 7.5 mAb14313 ND 1.1 1.55E10 7.1 mAb14314 ND 1.1 NC 3.9 mAb14315 ND 1.1 6.24E11 9.3 mAb14316 ND 1.0 1.89E10* 9.1 mAb1932 - ND 1.3 ND 1.5 Negative Control mAb2959 - 3.37E11 29.4 1.81E11 29.9 Positive Control *At high concentrations the signal decreased (hook effect); points on the hook were removed prior to EC50 determination NC: Not calculated because the data did not fit a 4-parameter logistic equation ND: Not determined because a concentration-dependent response was not observed Fold Induction = Max Average RLU within tested dose range of each antibody/Average RLU (background signal = no antibody)

[0288] From the initial mAbs tested, a select few were chosen for further follow-up to run alongside antibodies described in U.S. Pat. No. 10,787,501. The EC50 values and fold inductions are summarized in Table 26.

Results with Jurkat/hCD20/SARS-CoV-2 Spike Target Cells

[0289] Each of the newly purified SARS-CoV-2 spike antibodies showed greater fold induction than the negative control antibody. The EC50 values of the newly purified SARS-CoV-2 spike antibodies ranged from 188 pM to 575 pM, and the fold induction values ranged from 2.0 to 3.2. The positive CD20 control antibody showed a fold induction of 27.0 with an EC50 value of 51.8 pM.

Results with Jurkat/hCD20 Target Cells

[0290] Each of the newly purified SARS-CoV-2 spike antibodies behaved similarly to the negative control antibody showing minimal activity. By contrast, the positive CD20 control antibody showed a fold induction value of 23.5 with an EC50 value of 142 pM.

TABLE-US-00026 TABLE 26 Surrogate ADCC assay results Jurkat CD20 Jurkat/CD20/spike Fold Fold mAb # or Name EC50 (M) induction EC50 (M) induction mAb14255 ND 1.1 1.88E10 3.2 mAb14256 ND 1.1 5.75E10 2.0 mAb14257 ND 1.2 2.63E10 2.2 mAb14258 ND 1.2 2.06E10 2.7 mAb10984 ND 1.0 5.86E10 1.3 mAb10986 ND 1.0 NC 1.4 mAb10964 ND 1.2 1.11E10 1.8 mAb10977 ND 1.3 NC 4.1 mAb10933 ND 1.1 6.47E10 2.9 mAb10987 ND 1.0 4.71E10 4.5 mAb10989 ND 1.1 6.23E10 6.5 mAb1932 - ND 1.2 ND 1.2 Negative Control mAb2959 - 1.42E10 23.5 5.18E11 27.0 Positive Control Antibody described in U.S. Pat. No. 10,787,501 NC: Not calculated because the data did not fit a 4-parameter logistic equation. ND: Not determined because a concentration-dependent response was not observed Fold Induction = Max Average RLU within tested dose range of each antibody/Average RLU (background signal = no antibody)

[0291] A subsequent round of testing on newly-purified antibodies was also performed. The EC50 values and fold inductions are summarized in Table 27.

Results with Jurkat/hCD20/SARS-CoV-2 Spike Target Cells

[0292] Each of the newly purified anti-SARS-CoV-2-S antibodies showed greater fold induction than the negative control antibody. The EC50 values of the SARS-CoV-2 spike antibodies ranged from 81.7 pM to 905 pM, and the fold induction values ranged from 4.2 to 13.0. The positive CD20 control antibody showed a fold induction of 46.7 with an EC50 value of 74.4 pM.

Results with Jurkat/hCD20 Target Cells

[0293] All the newly purified SARS-CoV-2-S antibodies behaved similarly to the negative control antibody, showing minimal activity. By contrast, the positive CD20 control antibody showed a fold induction value of 62.1 with an EC50 value of 145 pM.

TABLE-US-00027 TABLE 27 Surrogate ADCC assay results Jurkat CD20 Jurkat/CD20/spike Fold Fold mAb # or Name EC50 (M) induction EC50 (M) induction mAb15156 ND 1.1 3.47E10 4.2 mAb15157 ND 1.3 3.98E10 4.8 mAb15158 ND 1.2 3.81E10 7.0 mAb15159 ND 1.0 4.17E10 5.0 mAb15160 ND 1.2 2.98E10 5.4 mAb15161 ND 1.2 4.94E10 5.9 mAb15162 ND 1.1 9.05E10 5.8 mAb15163 ND 1.1 3.42E10* 8.2 mAb15164 ND 1.2 2.23E10* 12.7 mAb15165 ND 1.1 2.01E10 13.0 mAb15166 ND 1.1 8.17E11 9.5 mAb15167 ND 1.0 1.11E10 7.3 mAb15170 ND 1.0 6.26E10 5.4 mAb15150 ND 1.1 4.59E10 5.2 mAb15151 ND 1.4 3.51E10 6.0 mAb14255.sup. ND 1.1 1.37E10 8.6 mAb14256.sup. ND 1.0 2.63E10 3.7 mAb14315.sup. ND 1.0 9.96E11* 11.5 mAb10985 ND 1.1 4.72E10 3.7 mAb10933 ND 1.0 8.76E11 7.7 mAb10987 ND 1.0 7.20E11 10.2 mAb1932 - ND 1.2 ND 3.7 Negative Control mAb2959 - 1.45E10 62.1 7.44E11 46.7 Positive Control Antibody described in U.S. Pat. No. 10,787,501 .sup.Antibody described in Table 25 and/or Table 26 *At high concentrations the signal decreased (hook effect); points on the hook were removed prior to EC50 determination ND: Not determined because a concentration-dependent response was not observed Fold Induction = Max Average RLU within tested dose range of each antibody/Average RLU (background signal = no antibody)

Example 9: Characterization of Anti-SARS-CoV-2-S mAbs in an ADCP Assay

[0294] The ability of antibodies targeting the spike protein of SARS-CoV-2 to induce phagocytosis of Jurkat cells engineered to express the SARS-CoV-2 full length spike protein was evaluated. Macrophages differentiated from monocytes in the presence of macrophage colony-stimulating factor (M-CSF) were used as effector cells in an antibody dependent cellular phagocytosis (ADCP) assay. An IgG1 isotype as a negative control was evaluated in parallel. Phagocytosis was assessed by measuring the number of fluorescently labeled target cells colocalizing with fluorescently labeled macrophages using an Opera Phenix High-Content Screening System.

Target Cells

[0295] Jurkat/hCD20/SARS-CoV-2 spike: Jurkat T cells were engineered to constitutively express full length human CD20 (amino acids M1-P297 of NCBI accession number NP_690605.1) as well as full-length SARS-CoV-2 spike protein (amino acids M1-T1273 of NCBI accession number YP_009724390.1). Jurkat/hCD20/SARS-CoV-2 spike cells were sorted for high expression of the spike protein and maintained in RPMI+10% FBS+P/S/G+1 g/ml puromycin+250 g/ml hygromycin growth medium.

Effector cells

[0296] Macrophages differentiated from monocytes in the presence of M-CSF: Frozen CD14+ monocytes (Lonza) were thawed, resuspended in assay media (RPMI 1640 supplemented with 10% FBS, 100 U/ml penicillin, 100 g/ml streptomycin, and 292 g/ml L-glutamine, NaPyr, HEPES, and NEAA), supplemented with 100 ng/ml M-CSF, and plated at 5.210.sup.4 cells/well into clear-bottom, collagen-coated 96-well black plates for differentiation into macrophages over 9 days.

Assay Set-Up

[0297] One day before the experiment, Jurkat target cells were split to 510.sup.5 cells/ml in their respective growth media.

[0298] On the day of the experiment, target cells and monocyte-derived macrophages were incubated in PBS supplemented with either CellTrace CFSE dye or CellTrace Violet dye, respectively, for 15 minutes at 37 C., 5% CO.sub.2.

[0299] CFSE-labeled target cells were washed and resuspended in assay media (RPMI 1640 supplemented with 10% FBS, 100 U/ml penicillin, 100 g/ml streptomycin, and 292 g/ml L-glutamine, NaPyr, HEPES, NEAA, and 10 M BME) and added in duplicate to 96-well U-bottom plates at a density of 510.sup.4 cells/well. Anti-SARS-CoV-2 spike and the IgG1 control antibodies were titrated in assay media in a 1:5 serial dilution ranging from 51 fM to 20 nM final concentration and added to the plates. The zero-antibody point was represented by 10 fM in GraphPad Prism.

[0300] After 15 minutes of incubation on ice the mixture of target cells, with or without titrated antibody, was then transferred to plates containing the violet-labelled macrophages and plates were incubated at 37 C., 5% CO2 for 30 minutes. Wells were washed with PBS 3 times, followed by addition of 4.21% formaldehyde in PBS supplemented with 2.5 M DRAQ5.

[0301] After a 20 minute incubation, wells were washed with PBS and imaged in both the 488 nm (CFSE-labelled target cells) and 375 nm (violet-labelled macrophages) excitation channels using an Opera Phenix High-Content Screening System. Image analysis was performed in Harmony software. Image segmentation in the 375 nm excitation channel was used to identify the macrophage population. Image segmentation in the 488 nm excitation channel was used to identify the target cells. Phagocytosis was quantified by identifying the macrophage population which contained target cells within them.

[0302] Percent phagocytosis was calculated by comparing number of macrophages undergoing phagocytosis to total macrophage cell number.

[00004]$\%\mathrm{ADCP}=\frac{\begin{array}{c}\mathrm{Violet}\mathrm{labelled}\mathrm{macrophages}\mathrm{containing}\\ \mathrm{CFSE}-\mathrm{labelled}\mathrm{target}\mathrm{cells}\end{array}}{\mathrm{Total}\mathrm{violet}\mathrm{labelled}\mathrm{macrophages}}$

[0303] For EC.sub.50 determinations, % ADCP was analyzed with GraphPad Prism using a 4-parameter logistic equation over a 10-point dose response curve, including the no antibody control. Maximum (Max) % ADCP was determined as the highest mean % ADCP value measured within tested dose range. The experiment was performed using a single donor in duplicate except for the following antibodies which were performed with no replicates: mAb10933, mAb10987, mAb10989, mAb1932, mAb10985, mAb14234, mAb10986, and mAb14259.

[0304] The EC.sub.50 values and maximum (Max) % ADCP are summarized in Table 28.

[0305] Results with Jurkat/hCD20/SARS-CoV-2 spike cells: Each of the 21 anti-SARS-CoV-2 spike antibodies showed greater Max % ADCP compared to IgG1 control antibody. The EC.sub.50 values of the SARS-CoV-2 spike antibodies ranged from 5.22 pM to 1.12 nM, and Max % ADCP ranged from 16.72% to 39.89%.

TABLE-US-00028 TABLE 28 ADCP Activity ADCP Antibody treatment Max % ADCP EC50 [M] mAb10933 33.12 3.88E11 mAb10987 36.30 3.19E11 mAb10989 33.02 4.82E11 mAb1932 7.69 ND mAb10985 31.80 2.17E10 mAb14234 32.84 7.27E11 mAb14249 32.60 1.69E10 mAb14255 21.17 1.73E11 mAb14256 19.50 NC mAb14287 33.24 NC mAb14288 25.86 6.14E10 mAb14290 16.72 NC mAb14291 32.85 4.12E10 mAb14295 35.30 4.57E10 mAb14296 31.60 5.07E10 mAb14312 27.77 1.69E11 mAb14314 16.72 1.12E09 mAb14315 34.80 6.30E11 mAb14316 37.72 2.54E10 mAb10964 34.03 5.22E12 mAb10986 39.89 7.13E11 mAb14259 39.56 NC Antibody described in U.S. Pat. No. 10,787,501 ND: Not determined because a concentration-dependent response was not observed. NC: Not calculated because the data did not fit a 4-parameter logistic equation. Max (% ADCP) is the highest mean % ADCP value within tested dose range.

Example 10: Structure Determination of Antibody-Bound Spike Protein and Interactions

[0306] To better understand the binding of mAb10987 and mAb14256 to the spike protein RBD, structural analysis was performed via cryo-electron microscopy (cryoEM). Fab fragments of mAb14256 and mAb10987 were isolated using FabALACTICA kit (Genovis). 25 g of the mAb14256 Fab and 25 g of mAb10987 Fab were mixed with 15 g of SARS-CoV-2-S RBD and incubated on ice for 30 min. For cryoEM grid preparation, the protein sample was diluted to 0.94 mg/mL and 0.15% PMAL-C8 amphipol was added. 3 L of protein was deposited onto a freshly plasma cleaned UltrAufoil grid (0.6/1.0, 300 mesh). Excess solution was blotted away using filter paper and plunge frozen into liquid ethane using a Vitrobot Mark IV. The cryoEM grid was transferred to a Titan Krios (Thermo Fisher) equipped with a K3 detector (Gatan). Movies were collected using EPU (Thermo Fisher) at 105,000 magnification, corresponding to a pixel size of 0.86 . A dose rate of 15 electrons per pixel per second was used and each movie was 2 seconds, corresponding to a total dose of 40 electrons per .sup.2.

[0307] All cryoEM data processing was carried out using cryoSPARC v3.2.0. 6,106 aligned micrographs were collected and 5,735 aligned micrographs were selected for further processing on the basis of estimated defocus values and CTF fit resolutions. An initial set of particles picked using blob picker were subjected to 2D classification to generate templates for template picking. 1.4 million particles picked by template picking were subjected to multiple rounds of 2D classification to remove unbound Fabs and particles containing an incomplete complex. Ab initio reconstruction with three classes generated two classes containing 301,461 particles that corresponded to the full mAb10987 FabmAb14256 FabRBD complex. Heterogeneous refinement of those two classes was performed where one class showed a good quality map while the other showed high preferred orientation. Non-uniform refinement of the particles in the good class containing 189,200 particles was followed by local refinement, resulting in a 3.86 resolution (FSC=0.143) map that was used for model building. Into this map, models of the RBD (taken from PDB code 6M0J) were manually placed and the two Fabs (taken from prior antibody structures, mAb10987 from PDB 6DG and mAb14256 from another mAb14256 model). These models were then manually rebuilt using Coot and real-space refined against the map using Phenix.

[0308] Confirming the above-described data, single-particle cryoEM of the complex of SARS-CoV-2 spike RBD bound to Fab fragments of mAb14256 and mAb10987 shows that the two antibodies in this cocktail can simultaneously bind to distinct regions of the RBD (FIG. 3, FIG. 4). A 3D reconstructed map of the complex with nominal resolution of 3.9 showed that both Fab fragments bound to non-overlapping regions on the surface of the RBD, confirming that they are non-competing antibodies. mAb14256 bound at the top of the RBD, extensively overlapping the binding site for ACE2. On the other hand, the epitope for mAb10987 was located on the side of the RBD, well away from the mAb14256 epitope, and had little to no overlap with the ACE2 binding site. The structure also revealed that mAb14256 competed with mAb10933 and mAb10985, although combinations of these antibodies still may be desirable because of different microepitope binding and the need to bind and neutralize different SARS-CoV-2 spike protein variants.

[0309] Using similar techniques, the structural basis for mAb15160 binding to SARS-CoV-2 RBD was investigated. Cryo-EM was used to determine the structure of the SARS-CoV-2 RBD in complex with the antigen-binding (Fab) fragments of mAb15160 and mAb14315. The resulting 3.18 -resolution structure revealed that mAb15160 binds to a small epitope on the top (i.e., closest to the target cell membrane) surface of the SARS-CoV-2 RBD (FIG. 5). Analysis of the binding interface between the SARS-CoV-2 RBD and the antibody Fab fragment indicate that the mAb15160 Fab contacts the RBD with both its heavy chain (13 residue-residue interactions) and light chain (13 residue-residue interactions). A total of 13 SARS-CoV-2 RBD residues are involved in mAb15160 binding. The functional impact of K417N, E484A, and Q493R, all of which are present in the Omicron variant, as well as other substitutions within the binding epitope, was assessed in neutralization studies discussed above. mAb15160 retained neutralization potency against the K417N and E484A individual mutations. There was an impact on the neutralization potency against the Q493R individual mutation (5-fold reduction compared with reference pseudoparticles); however, the neutralization potency of mAb15160 was within 1.5-fold against pseudoparticles pseudotyped with the full S protein sequence from the BA.1, BA.1.1, and BA.2 (Omicron) lineages compared with reference pseudoparticles.

[0310] Using similar techniques, the structural basis for mAb14284 binding to SARS-CoV-2 Wuhan-Hu-1 and BA.1 (omicron lineage) RBDs was investigated. Cryo-EM was used to determine the structure of the antigen-binding (Fab) fragments of mAb14284 and mAb15160 with the RBD from the Wuhan-Hu-1 strain and BA.1 lineage. The resulting 3.3 and 3.4 -resolution structures, respectively, revealed that mAb14284 binds to an upper patch on both the Wu-1 and BA.1 RBD in a manner that is competitive with ACE2 binding (FIG. 6). Analysis of the binding interface between the mAb14284 Fab fragment and the SARS-CoV-2 RBDs from Wu-1 and BA.1 indicate that mAb14284 binds to the same location on the Wu-1 and BA.1 RBDs, with the same epitope residues except for 4 changes located at the periphery (Table 29, below). Epitope residues mutated in the BA.1 variant (N440K, G446S, Q498R, N501Y) relative to the ancestral Wu-1 strain are accommodated in the structure without major changes at the antibody-antigen interface.

TABLE-US-00029 TABLE 29 Summary of Interactions between mAb14284 Fab Residues and SARS-CoV-2 RBD Residues Wuhan- mAb14284 Residue(s) Hu-1 mAb 14284 Residue(s) Interacting BA.1 Interacting with Indicated Type of RBD with Indicated RBD Residue RBD RBD Residue Interaction Residue Heavy Chain Light Chain Residue Heavy Chain Light Chain Hydrogen Asn439 Tyr34 bond/salt Lys444 Trp55, Asp56 Lys444 Trp55 bridge (<4 ) Val445 Arg60 Gly447 Arg60 Gly447 Arg60 Gln498* Thr95 Arg498* Thr95 Thr500 Val31 Hydrophobic Asn440* Ile102, Pro103 Lys440* Ile102, Pro103 (<5 ) Leu441 Ser32, Gly33 Leu441 Ser32 Ser443 Ile102 Ser443 Ile102 Lys444 Tyr54, Trp55, Lys444 Tyr54, Trp55, Asp56, Asp58, Asp58, Ile102 Arg60, Ile102 Val445 Trp49, Leu52, Tyr93, Ser97, Thr98 Val445 Trp49, Leu52, Tyr93, Ser97, Tyr54, Arg60, Tyr54, Arg60, Thr98 His100 His100 Gly446* Arg60 Ser96 Ser446* Arg60 Ser96 Gly447 Arg60 Gly447 Arg60 Tyr449 Arg60 Gln498* Thr95 Arg498* Thr95 Pro499 Ile102 Tyr34,Tyr93 Pro499 Ile102 Tyr34,Tyr93 Thr500 Leu29, Gly30, Tyr93, Thr500 Leu29, Thr94, Thr94 Thr95 Asn501* Val31 Tyr501* Val31 Gly502 Val31 Gly502 Val31 SARS-CoV-2 Wuhan-Hu-1 and BA.1 S protein RBD residues that interact with residues of the mAb14284 Fab heavy chain and light chain are indicated. A hyphen indicates that no interaction was observed between the indicated RBD residue and the indicated antibody chain. Residues were selected based on the presence of hydrogen bonds/salt bridges with interatomic distances <4 (not including hydrogen atoms) or hydrophobic surfaces with interatomic distances <5 . An asterisk *indicates an RBD residue mutated in the BA (Omicron) lineages.

[0311] To assess the degree of variability of the mAb14284 epitope across currently circulating SARS-CoV-2 viruses, the sequences of 3,825,870 publicly available SARS-CoV-2 genomes identified between 1 Jan. 2022 and 16 Jun. 2022 (i.e., during the Omicron wave) were utilized. The sequences collected during this period likely consist mostly of Omicron sequences, however, other circulating variants, such as Delta, may also be included in this analysis.

[0312] Between 1 Jan. 2022 and 16 Jun. 2022, the RBD residues bound by mAb14284 were 99.96% conserved relative to the Wuhan-Hu-1 strain at each position, with the exception of 4 residues, N440, G446, Q498, and N501, which are mutated in the Omicron variant (i.e., BA lineages) (Table 30). The frequencies of N440, G446, Q498, and N501 across this monitoring period were 22, 59, 13, and 12%, respectively; instead, N440K, Q498R, and N501Y, all of which are present in the Omicron variant, had become predominant. While G446 remains the predominant residue during this period, 39% of sequences contain the G446S mutation, which is present in the BA.1 lineage of the Omicron variant

[0313] The functional impact of N440K, G446S, Q498R, and N501Y, as well as other substitutions within the binding epitope identified during the conservation analysis (specified below in Table 30) was assessed in neutralization studies as discussed in Example 4. mAb14284 retained neutralization potency against all epitope residue variants tested except mutations of K444. The K444L, K444M, K444N, K444Q, and K444T individual substitutions resulted in a >5-fold change compared with reference pseudoparticles; however, the neutralization potency of mAb14284 was within 1.5-fold against pseudoparticles pseudotyped with the most predominant (ie, 0.0141%) K444 variant, K444R (Table 19).

TABLE-US-00030 TABLE 30 Summary of Frequencies of REGN14284 Epitope Variants Across Publicly Available SARS-CoV-2 Genomes from 1 Jan. 2022 through 16 Jun. 2022 Observed Order of Reference Frequency for Epitope Variants AA Sequence (High to Low) Position (Wuhan-Hu-1) 1.sup.st 2.sup.nd in RBD Residue Freq (%) Residue Freq (%) Residue Freq (%) 440 N 22.3472 K 78.1540 del 0.0026 441 L 99.9942 del 0.0025 F 0.0016 443 S 99.9964 del 0.0024 N 0.0002 444 K 99.9699 R 0.0141 N 0.0095 445 V 99.9834 A 0.0082 I 0.0033 446 G 58.9933 S 38.6232 V 0.0133 447 G 99.9948 del 0.0033 S 0.0005 449 Y 99.9643 H 0.0199 N 0.0137 498 Q 12.7255 R 87.5676 L 0.0115 499 P 99.9840 del 0.0095 R 0.0025 500 T 99.9832 del 0.0097 S 0.0031 501 N 12.3329 Y 87.9706 del 0.0101 502 G 99.9853 del 0.0104 L 0.0007 A total of 3,825,870 publicly available SARS-CoV-2 genome sequences identified Jan. 1, 2022 through Jun. 16, 2022 were analyzed against the reference (Wuhan-Hu-1) SARS-CoV-2 genome. The data collected during this period likely reflects mostly Omicron sequences, however, other circulating variants, such as Delta, may also be included in this analysis. The frequency (freq) of variants within the mAb14284 epitope (as determined by structural studies described herein) was calculated. The amino acid (AA) position indicates the residue in the SARS-CoV-2 S protein RBD (as displayed in Table 29). For each AA position within the epitope, the frequencies of reference residues and the 2 most frequent variant residues are shown. Del indicates a deletion at that AA position. The functional impact of the most predominant circulating epitope residue variants assessed through Jun. 16, 2022 (N439K, N439V, N440D, N440K, L441F, L441Q, K444L, K444M, K444N, K444Q, K444R, K444T, V445A, V445T, G446D, G446R, G446S, G446V, G447V, N448S, Y449D, Y449F, Y449H, Y449N, Y449S, Q498H, Q498R, T500A, T500F, T500N, N501T, N501Y) is summarized in Table 19.

Example 11: In Vitro VSV-SARS-CoV-2-S Virus Escape Mutant Selection in the Presence of mAb15160

[0314] To assess SARS-CoV-2 escape mutant selection, Vero cells were infected with replicating VSV-SARS-CoV-2-S (Wuhan-Hu-1 strain), and incubated with a range of concentrations of mAb15160 or IgG1 isotype control for 4 days. Infected cells were monitored for virus-induced cytopathic effect (CPE) as a read-out for virus replication. Loss of neutralization efficacy, as assessed by observable CPE in the presence of increasing concentration of antibodies, indicated potential selection of escape mutants. Supernatants containing virus were collected from wells with the highest mAb concentration and observable CPE, and further passaged in the presence of the same mAb. After passaging, supernatants containing virus were subjected to RNA sequence analysis and all amino acid changes in the S protein were designated as putative escape mutations if the amino acid change was present in anti-SARS-CoV-2 S protein mAb-treated samples, but neither in input virus (inoculum) nor control samples (IgG1 control or no-mAb control).

[0315] CPE at 90% was observed in the presence of 0.016 to 50 g/mL mAb15160 within the first passage. Deep sequencing of viral genomes after the first passage under selection with mAb15160 identified 2 putative escape mutations in the SARS-CoV-2 RBD binding epitope for mAb15160F486V and Y489H. Of note, neither of these individual mutations are found in the prominent Omicron and Delta variants or in any other prior variant of concern, indicating that there is no current clinical concern with respect to these mutations.

[0316] The functional impact of each putative escape mutant on mAb15160 activity was assessed in neutralization assays, as discussed above. F486V and Y489H reduced the neutralization activity of mAb15160 by >531-fold and 122-fold, respectively, compared with reference pseudoparticles, confirming that F486V and Y489H are escape mutants of mAb15160.

Example 12: In Vitro Effector Function Activities of mAb15160

[0317] The ability of mAb15160 to mediate antibody dependent cellular phagocytosis (ADCP) of fluorescently labeled target cells engineered to express full length (FL) SARS-CoV-2 S protein (Jurkat/hCD20/SARS-CoV-2 S FL) was assessed in the presence of fluorescently-labeled primary MDM effector cells from two independent donors that were differentiated with macrophage colony-stimulating factor (MCSF). Results demonstrated that mAb15160 mediated concentration-dependent ADCP of Jurkat/hCD20/SARS-CoV-2 S FL cells with EC.sub.50 values in the picomolar range (Table 31), while no ADCP was observed with parental cells that did not express SARS-CoV-2 S protein. An IgG1 isotype control did not mediate ADCP of either Jurkat/hCD20/SARS-CoV-2 S FL or Jurkat/hCD20 cell lines, whereas an anti-CD20 IgG1 control mediated ADCP of both Jurkat/hCD20/SARS-CoV-2 S FL and Jurkat/hCD20 cell lines, demonstrating the capacity of effector cells used in the assay to phagocytose each of the tested cell lines.

TABLE-US-00031 TABLE 31 Summary of EC.sub.50 and Maximum Phagocytosis Values for ADCP of Target Cells Mediated by mAb15160 MDM Donor #1 Jurkat/hCD20/ SARS-CoV- MDM Donor #2 2 S FL Jurkat/hCD20 Jurkat/hCD20/ Jurkat/hCD20 Max Max SARS-CoV-2 S FL Max EC.sub.50 ADCP.sup.a EC.sub.50 ADCP.sup.a EC.sub.50 Max EC.sub.50 ADCP.sup.a mAb(s) (M) (%) (M) (%) (M) ADCP.sup.a (%) (M) (%) mAb15160 7.81E11 27.5 ND 14.7 4.76E11 31.1 ND 11.1 mAb10933 9.70E12 28.3 ND 14.0 2.23E11 28.6 ND 11.5 (casirivimab) mAb10987 2.91E11 32.3 ND 14.8 1.98E11 30.4 ND 10.7 (imdevimab) casirivimab + imdevimab 1.20E10 49.5 ND 15.5 5.78E11 48.2 ND 15.1 IgG1 isotype control ND 13.1 ND 16.9 ND 9.6 ND 14.6 Anti-CD20 IgG1 1.01E09 31.1 7.75E10 45.4 1.49E10 30.3 4.67E10 45.2 .sup.aMaximum (max) ADCP is defined as the highest mean percentage ADCP value observed within the concentration range tested (0 nM to 20 nM, where 0 nM is the no-antibody control). ND, not determined because a concentration-dependent increase was not observed

[0318] mAb15160 was evaluated for the ability to mediate antibody dependent cell-mediated cytotoxicity (ADCC) against Jurkat/hCD20/SARS-CoV-2 S FL and Jurkat/hCD20 target cells using human primary natural killer (NK) cells from three independent donors as effector cells. Results demonstrate that mAb15160 mediates ADCC against Jurkat/hCD20/SARS-CoV-2 S FL target cells, but not SARS-CoV-2-negative parental cells, with varying activity demonstrated with each NK donor (donor #3>donor #2; donor #1 showed no activity (Table 32). An IgG1 control did not mediate ADCC of either Jurkat/hCD20/SARS-CoV-2 S FL or Jurkat/hCD20 cell lines, whereas a positive control, anti-CD20 IgG1, mediated ADCC of both Jurkat/hCD20/SARS-CoV-2 S FL and Jurkat/hCD20 cell lines, demonstrating cytotoxic potential of all NK cells used in the assay against each tested cell line.

TABLE-US-00032 TABLE 32 Summary of EC.sub.50 and Maximum Cytotoxicity Values for ADCC Against Target Cells Mediated by mAb15160 NK Donor #1 NK Donor #2 NK Donor #3 Jurkat/hCD20/ Jurkat/hCD20/ Jurkat/hCD20/ SARS-CoV-2 S SARS-CoV-2 S SARS-CoV-2 S FL Jurkat/hCD20 FL Jurkat/hCD20 FL Jurkat/hCD20 Max Max Max Max Max Max Cyto- Cyto- Cyto- Cyto- Cyto- Cyto- EC.sub.50 toxicity.sup.a EC.sub.50 toxicity.sup.a EC.sub.50 toxicity.sup.a EC.sub.50 toxicity.sup.a EC.sub.50 toxicity.sup.a EC.sub.50 toxicity.sup.a mAb(s) (M) (%) (M) (%) (M) (%) (M) (%) (M) (%) (M) (%) mAb15160 ND 1.8 NC 4.6 NC 6.3 ND 3.1 6.32E 12.9 ND 0.0 11 mAb10933 ND 1.4 NC 4.7 NC 5.0 NC 3.6 4.89E 11.1 ND 1.2 (casirivimab) 11 mAb 10987 7.42E 8.1 NC 3.9 5.05E 10.5 ND 1.9 6.44E 26.6 ND 1.3 (imdevimab) 10 09 11 casirivimab + ND 1.6 ND 0.8 NC 4.1 ND 4.8 3.60E 15.4 ND 0.0 imdevimab 11 IgG1 ND 1.8 ND 0.7 ND 0.0 NC 6.4 ND 0.0 ND 0.5 isotype control Anti-CD20 2.63E 20.0 3.03E 21.0 8.19E 15.1 8.08E 26.8 3.53E 14.5 NC 37.7 IgG1 12 12 12 13 14 .sup.aMaximum (max) cytotoxicity is defined as the highest mean percentage cytotoxicity value observed within the concentration range tested NC, Not calculated because the data did not fit a 4-parameter logistic equation; ND, Not determined because a concentration-dependent increase was not observed.

[0319] mAb15160 was evaluated for the ability to mediate the activation of FCGR3A signaling in a surrogate ADCC reporter bioassay using Jurkat T cells engineered to express a nuclear factor of activated T cells-luciferase (NFAT-Luc) reporter gene and human FCGR3A (Jurkat/NFAT-Luc/FCGR3A) in the presence of Jurkat/hCD20/SARS-CoV-2 S FL target cells. Engagement of FCGR3A via the Fc domain of human IgG1 antibodies bound to target cells leads to the activation of NFAT and drives the expression of luciferase, which is then measured via a luminescence readout. In agreement with results from ADCC assays using primary NK cells, results from the ADCC-surrogate reporter assay demonstrate that mAb15160 mediates a concentration-dependent increase in luciferase activity from reporter cells expressing FCGR3A in the presence of Jurkat/hCD20/SARS-CoV-2 S FL target cells with an EC.sub.50 of 2.9710.sup.10 M. The IgG1 negative control did not activate FCGR3A in the presence of Jurkat/hCD20/SARS-CoV-2 S FL or Jurkat/hCD20 cell lines.

[0320] mAb15160 was evaluated for the ability to mediate CDC against Jurkat/hCD20/SARS-CoV-2 S FL and Jurkat/hCD20 target cells in the presence of 5% pooled (ie, from several donors) NHS. Results demonstrate that mAb15160 does not mediate CDC against Jurkat/hCD20/SARS-CoV-2 S FL or Jurkat/hCD20 cells in the presence of 5% NHS at antibody concentrations ranging from 0.95 pM to 1 M. NHS used in the assay was evaluated in the same target cell lines for the ability to mediate CDC using the positive control, anti-CD20 IgG1. In the presence of NHS, the anti-CD20 IgG1 mediated CDC against target cell lines in a concentration-dependent manner. In the absence of NHS, the anti-CD20 IgG1 did not mediate lysis against any of the tested target cell lines.

Example 13: In Vitro VSV-SARS-CoV-2-S Virus Escape Mutant Selection in the Presence of mAb14284

[0321] To assess SARS-CoV-2 escape mutant selection, Vero cells were infected with replicating pseudotyped VSV-SARS-CoV-2-S (Wuhan-Hu-1 strain) and incubated with a range of concentrations of mAb14284 or IgG1 isotype control for 4 days. Infected cells were monitored for virus-induced cytopathic effect (CPE) as a read-out for virus replication. Loss of neutralization efficacy, as assessed by observable CPE in the presence of increasing concentration of antibodies, indicated potential selection of escape mutants. Supernatants containing virus were collected from wells with the highest mAb concentration and observable CPE, and further passaged in the presence of the same mAb. After passaging, supernatants containing virus were subjected to RNA sequence analysis and all amino acid changes in the S protein were designated as putative escape mutations if the amino acid change was present in the mAb14284-treated sample, but not in the IgG1 control sample.

[0322] CPE at 90% was observed in the presence of 0.016 to 50 g/mL mAb14284 within the first passage (Table 33). Deep sequencing of viral genomes after the first passage under selection with mAb14284 identified 3 putative escape mutations in the SARS-CoV-2 RBD binding epitope for REGN14284-K444E, K444N, and V445D (Table 34).

[0323] The functional impact of putative escape mutant, K444N, on mAb14284 activity was assessed in neutralization assays discussed in Example 4. The K444N variant resulted in a complete loss of mAb14284 neutralization activity compared with reference pseudoparticles (Table 19), confirming that K444N is an escape mutant of mAb14284.

[0324] Functional assays of K444E and V445D are ongoing. However, the K444L, K444M, K444N, K444Q, K444T, V445A, and V445T mutations all resulted in a reduction or complete loss of mAb14284 neutralization activity; mAb14284 neutralization activity was retained against the K444R mutation (Table 19).

TABLE-US-00033 TABLE 33 Selection of Putative VSV-SARS-CoV-2-S Virus Escape Mutants CPE (%) Antibody 50 g/mL 10 g/mL 2 g/mL 0.4 g/mL 0.08 g/mL 0.016 g/mL No mAb Passage 1 mAb14284 [00002] >90 >90 >90 >90 >90 >90 IgG1 Control [00003] >90 >90 >90 >90 >90 [00004] Vero cells were infected with VSV-SARS-CoV-2-S virus in the presence of mAb14284 or IgG1 isotype control mAb at a range of concentrations (0.016 to 50 g/mL total) for 4 days. Cells were screened for virus replication by monitoring for virus-induced CPE. Supernatants containing virus were collected from wells with the highest antibody concentrations and detectable viral replication (20% CPE). Deep sequencing of viral RNA from was performed to determine the identity of putative escape mutations. A no antibody control (0 g/mL) was sequenced to monitor for tissue culture adaptation. Bold, underlined text indicates samples that were sequenced.

TABLE-US-00034 TABLE 34 Deep Sequencing of Passaged Virus Identifies Putative Escape Mutations in the Receptor Binding Domain Genomic position 4407 4409 4411 Reference nucleotide A G T Variant nucleotide G T A Gene position 1330 1332 1334 Mutation nature nSNP nSNP nSNP AA position 444 444 445 Reference AA K K V Variant AA E N D Inoculum 0% 0% 0% mAb14284 7% 19% 67% IgG1 Isotype Control 0% 0% 0% No Ab 0% 0% 0% Viral RNA from wells with the highest mAb concentration that showed detectable CPE was isolated 4 days post-infection. Deep sequencing was performed to identify changes in S protein sequence relative to the Wuhan-Hu-1 reference sequence. Only RBD variants are summarized in this table.

Example 14: In Vitro Effector Function Activities of mAb14284

[0325] IgG antibodies that are bound to target cells can mediate effector functions via their constant (Fc) region by interacting with specific FCGRs expressed on macrophages and natural killer (NK) cells, leading to antibody-dependent cellular phagocytosis (ADCP) and antibody-dependent cellular cytotoxicity (ADCC), respectively, or by binding to complement proteins, leading to complement-dependent cytotoxicity (CDC), resulting in the destruction of target cells.

[0326] A series of in vitro studies was conducted to assess the ability of mAb14284 to mediate ADCP, ADCC, and CDC against target cells expressing full-length (FL) SARS-CoV-2 S protein from the Wuhan-Hu-1 strain (AA 1-1273), and BA.1 and BA.2 omicron lineages (AA 1-1270). To allow for simultaneous testing of an anti-CD20 IgG1 positive control antibody, target cell lines were also engineered to stably express human CD20 (hCD20).

[0327] Specifically, mAb14284 was assessed for the ability to: 1) mediate ADCP of target cells using monocyte-derived macrophages (MDM) as effector cells; 2) mediate ADCC of target cells using primary natural killer (NK) cells as effector cells; 3) activate FCGR3A signaling in an ADCC-surrogate reporter assay; and 4) mediate CDC of target cells in the presence of normal human serum (NHS).

[0328] The ability of mAb14284 to mediate ADCP of fluorescently labeled Jurkat/hCD20 target cells expressing FL Wu-1, BA.1, or BA.2 S protein was assessed in the presence of fluorescently labeled primary MDM effector cells from two independent donors that were differentiated with macrophage colony-stimulating factor (MCSF). mAb14284 mediates concentration-dependent ADCP of Jurkat/hCD20 cells expressing SARS-CoV-2 Wu-1, BA.1, or BA.2 S protein with EC.sub.50 values in the picomolar range (Table 35), while no ADCP was observed with parental cells that do not express SARS-CoV-2 S protein. An IgG1 isotype control did not mediate ADCP of any of the tested cell lines, whereas an anti-CD20 IgG1 control mediated ADCP of all tested cell lines, demonstrating the capacity of effector cells used in the assay to phagocytose each cell line evaluated.

TABLE-US-00035 TABLE 35 Summary of EC.sub.50 and Maximum Phagocytosis Values for ADCP of Target Cells Mediated by mAb14284 Jurkat/hCD20/SARS- Jurkat/hCD20/SARS- Jurkat/hCD20/SARS- CoV-2 Wu-1 S CoV-2 BA.1 S CoV-2 BA.2 S Jurkat/hCD20 Max ADCP.sup.a Max ADCP.sup.a Max ADCP.sup.a Max ADCP.sup.a mAb(s) EC.sub.50 (M) (%) EC.sub.50 (M) (%) EC.sub.50 (M) (%) EC.sub.50 (M) (%) MDM Donor #1 mAb14284 NC 38.6 3.11E11 44 4.37E11 46.5 ND 0 IgG1 isotype 8.78E11 45.5 7.97E11 61.5 2.30E10 37.4 1.77E10 54.9 control Anti-CD20 IgG1 ND 2 ND 4.4 ND 4.1 ND 4.5 MDM Donor #2 mAb14284 1.64E11 49.7 3.73E11 60.9 8.74E11 43.8 ND 0 IgG1 isotype 8.10E11 87.3 8.21E11 80.4 2.23E10 22.5 3.18E10 37.9 control Anti-CD20 IgG1 ND 7.4 ND 1.5 ND 0 ND 0 .sup.aMaximum (max) ADCP is defined as the highest mean percentage ADCP value observed within the concentration range tested (0 nM to 20 nM, where 0 nM is the no-antibody control). NC, Not calculated because the data did not fit a 4-parameter logistic equation; ND, not determined because a concentration-dependent increase was not observed

[0329] mAb14284 was evaluated for the ability to mediate ADCC against Jurkat/hCD20 target cells expressing FL Wu-1, BA. 1, or BA.2 S protein using human primary NK cells pooled from 3 donors as effector cells. Results demonstrate that mAb14284 mediates concentration-dependent ADCC against Jurkat/hCD2S expressing SARS-CoV-2 Wu-1, BA.1, or BA.2 S protein with EC.sub.50 values in the picomolar range (Table 36), but not against parental cells that do not express SARS-CoV-2 S protein. An IgG1 control did not mediate ADCC against any of the tested cell lines, whereas a positive control, anti-CD20 IgG1, mediated ADCC against all tested cell lines, demonstrating cytotoxic potential of the NK cells used in the assay against each cell line evaluated.

TABLE-US-00036 TABLE 36 Summary of EC.sub.50 and Maximum Cytotoxicity Values for ADCC Against Target Cells Mediated by REGN14284 Jurkat/hCD20/ Jurkat/hCD20/ Jurkat/hCD20/ SARS-CoV-2 Wu-1 S SARS-CoV-2 BA.1 S SARS-CoV-2 BA.2 S Jurkat/hCD20 Max Max Max Max Cytotoxicity.sup.a Cytotoxicity.sup.a Cytotoxicity.sup.a Cytotoxicity.sup.a mAb(s) EC.sub.50 (M) (%) EC.sub.50 (M) (%) EC.sub.50 (M) (%) EC.sub.50 (M) (%) mAb14284 6.41E12 7.9 2.64E11 8.3 5.37E12 8.3 ND 0.4 IgG1 isotype ND 0.6 ND 0.2 ND 0.6 ND 0.6 control Anti-CD20 IgG1 5.83E12 16.9 3.03E12 18.8 1.49E11 18.7 1.17E11 15.1 .sup.aMaximum (max) cytotoxicity is defined as the highest mean percentage cytotoxicity value observed within the concentration range tested NC, Not calculated because the data did not fit a 4-parameter logistic equation; ND, Not determined because a concentration-dependent increase was not observed.

[0330] mAb14284 was evaluated for the ability to mediate the activation of FCGR3A signaling in a surrogate ADCC reporter bioassay using Jurkat T cells engineered to express a nuclear factor of activated T cells-luciferase (NFAT-Luc) reporter gene and human FCGR3A (Jurkat/NFAT-Luc/FCGR3A) in the presence of Jurkat/hCD20 target cells expressing FL SARS-CoV-2 Wu-1, BA.1, or BA.2 S protein. Engagement of FCGR3A via the Fc domain of human IgG1 antibodies bound to target cells leads to the activation of NFAT and drives the expression of luciferase, which is then measured via a luminescence readout.

[0331] In agreement with results from ADCC assays using primary NK cells, results from the ADCC-surrogate reporter assay demonstrate that mAb14284 mediates a concentration-dependent increase in luciferase activity from reporter cells expressing FCGR3A in the presence of SARS-CoV-2 Wu-1, BA.1, or BA.2 S protein-expressing target cells with picomolar EC.sub.50 values, but not against parental cells that do not express SARS-CoV-2 S protein. The IgG1 negative control did not activate FCGR3A in the presence of any cell line evaluated.

[0332] mAb14284 was evaluated for the ability to mediate CDC against Jurkat/hCD20/SARS-CoV-2 Wu-1 S, Jurkat/hCD20/SARS-CoV-2 BA.1 S, Jurkat/hCD20/SARS-CoV-2 BA.2 S, and Jurkat/hCD20 target cells in the presence of 5% pooled (i.e., from several donors) NHS. Results demonstrate that mAb14284 did not mediate CDC against parental Jurkat/hCD20 cells, nor Jurkat/hCD20 cells expressing full-length SARS-CoV-2 Wu.1, BA.1, or BA.2 S protein in the presence of 5% NHS at antibody concentrations ranging from 0.95 pM to 1 M.

[0333] NHS used in the assay was evaluated in the same target cell lines for the ability to mediate CDC using the positive control, anti-CD20 IgG1. In the presence of NHS, the anti-CD20 IgG1 mediated CDC against target cell lines in a concentration-dependent manner. In the absence of NHS, the anti-CD20 IgG1 did not mediate lysis against any of the tested target cell lines.

Example 15: Phase 3 Clinical Trial Assessing the Safety and Efficacy of mAb15160

[0334] After obtaining safety and tolerability data from healthy volunteers treated with mAb15160, a phase 3, randomized, double-blind, placebo-controlled study is initiated to assess the safety and efficacy of mAb15160 to prevent symptomatic COVID-19 (Table 37). This is an event-driven study that is anticipated to enroll approximately 5000 participants of age 18 years with and without risk factors for severe SARS-CoV-2 infection, who are PCR negative for SARS-CoV-2 at the time of randomization. Study participants are randomized in a 1:1 ratio into one of 2 treatment groups (300 mg mAb15160 administered once every 12 weeks (Q12W) or placebo) administered as subcutaneous (SC) injections over 6 months.

[0335] The primary endpoint of this study is symptomatic RT-qPCR confirmed SARS-CoV-2 infection. The secondary endpoints include the safety and tolerability of repeated SC injections of mAb15160. Safety assessments include, but are not limited to, collection of treatment emergent adverse events, vital signs, and safety laboratory tests. Blood is also drawn for drug concentration, immunogenicity, and serum neutralizing titer measurements. Administration of mAb15160 results in a reduction in the percentage of symptomatic SARS-CoV-2 infection as compared to the placebo group.

TABLE-US-00037 TABLE 37 Phase 3 study design Hypothesis: Prophylaxis with mAb15160 prevents symptomatic SARS-CoV-2 infection. Treatment Arms and Dose: mAb15160 (300 mg SC Q12W 2 doses) Placebo SC Key Design Elements This is an event-driven, phase 3, randomized, double-blind, placebo-controlled study to assess the safety and efficacy of mAb15160 to prevent symptomatic COVID-19 This study enrolls approximately 5000 participants of age 18 years with and without risk factors for severe SARS-CoV-2 infection. Study participants will be randomized in a 1:1 ratio into one of 2 treatment groups (mAb15160, or placebo) administered as subcutaneous injections over 6 months. Duration: 253 days Population: 18 years-old with a SARS-CoV-2 negative RT-qPCR with and without risk factors for severe COVID-19 Number of subjects Approximately 5000 participants planned: Primary Endpoint: Symptomatic RT-qPCR-confirmed SARS-CoV-2 infection cases during the 6- month efficacy assessment period Secondary Endpoint: Safety and tolerability of repeated SC injections of mAb15160 Serum concentration and Immunogenicity of mAb15160 Q12W = dosed once every 12 weeks

Example 16: Phase 3 Clinical Trial Assessing the Safety and Efficacy of mAb15160 in Pre-Exposure Prophylaxis

[0336] This is a phase 3, randomized, double-blind, placebo-controlled study in adults and adolescents to assess the safety and efficacy of mAb15160 as pre-exposure prophylaxis to prevent COVID-19. This event-driven study enrolls approximately 5000 participants of age >12 years. Study participants is randomized in a 1:1 ratio into one of 2 treatment groups (mAb15160 or placebo) administered as subcutaneous injections over 6 months. The study consists of three periods: a 1 to 14-day screening period, a 6-month prophylaxis and efficacy assessment period (EAP), and a 3-month follow-up period (see Table 38). Administration of mAb15160 results in a reduction in the percentage of symptomatic SARS-CoV-2 infection as compared to the placebo group.

Screening/Baseline

[0337] After participants provide informed consent, they are assessed for study eligibility. The screening visit occurs up to 14 days prior to the randomization and baseline visit, and in some subjects the screening, randomization and baseline visit may occur on the same day. To be included in the study, participants have a negative SARS-CoV-2 test result, preferably from a reverse-transcriptase polymerase-chain-reaction (RT-PCR) assay from a sample collected within 72 hours of the baseline visit. Randomization will be stratified for assignment of treatment group by age group category (12 years to <18 years, 18 years to 65 years, >65 years), immunocompromised status (yes/no), and by prior vaccination (yes/no). Approximately 20% or more of participants enrolled will have a status of immunocompromised.

Treatment and Efficacy Assessment Period (EAP)

[0338] On day 1, after assessing available lab results, completing baseline assessments, sample collection, and randomization, all participants receive their first dose of study drug. Study drug is administered as a 2.5 ml subcutaneous (SC) injection in a blinded manner. Participants are randomized in a 1:1 ratio to one of the two treatment groups: mAb15160 (300 mg administered subcutaneously once every 8 weeks (SC Q8W) or placebo administered subcutaneously once every 8 weeks.

TABLE-US-00038 TABLE 38 Phase 3 study design HYPOTHESIS Prophylaxis with subcutaneous mAb15160 will prevent symptomatic SARS-CoV-2 infection. Note: Symptomatic SARS-CoV-2 infection is determined by a positive SARS- CoV-2 RT-qPCR result during the Efficacy Assessment Period (EAP) with at least one symptom occurring within 10 days of a positive RT-qPCR. The EAP is from baseline to 6 months. STUDY OBJECTIVES Primary Objective: Primary Endpoint: AND ENDPOINTS To evaluate the efficacy of Cumulative incidence of mAb15160, compared with symptomatic, RT-PCR-confirmed placebo, in preventing SARS-CoV-2 infection cases during symptomatic SARS-CoV-2 the efficacy assessment period (EAP) infection. Secondary Objectives: Secondary Endpoints: To evaluate the safety and Proportion of participants with tolerability of repeated SC treatment-emergent adverse events injections of mAb15160 in the (TEAEs), during the EAP and study population follow-up period To assess concentrations of Proportion of participants with mAb15160 in serum over time TEAEs leading to study drug To assess the immunogenicity discontinuation during the EAP and of mAb15160 follow-up period Proportion of participants with treatment-emergent serious adverse events (SAEs) during the EAP and follow-up period Proportion of participants with adverse events of special interest (AESIs) during the EAP Concentrations of mAb15160 in serum over time Incidence and titer of anti-drug antibodies (ADA), and incidence of neutralizing antibodies (NAb) to mAb15160 over time Exploratory Objectives: Exploratory Endpoints: To evaluate additional Cumulative incidence of indicators of mAb15160 symptomatic, RT-PCR-confirmed clinical efficacy and disease SARS-CoV-2 infection cases during prevention, compared to the follow-up period placebo Cumulative incidence of RT-PCR- To assess humoral immune confirmed SARS-CoV-2 infection response to SARS-CoV-2 at cases (regardless of symptoms) baseline and after during the EAP administration of mAb15160 Cumulative incidence of To assess serum neutralization symptomatic, RT-PCR-confirmed potential of mAb15160 SARS-CoV-2 infection cases during To characterize viral variants the efficacy assessment period by sequencing SARS-CoV-2 in (EAP), in subgroups of those with participants who become one or more risk factors for severe infected post-baseline COVID To explore biomarkers Proportion of participants with 1 predictive or indicative of moderate COVID-19 symptom safety and/or efficacy of during the EAP and follow-up period mAb15160, COVID-19 Proportion of participants with vaccine response, SARS-CoV- COVID- 19-related hospitalization, 2 infection and immune emergency room visit, urgent care response, COVID-19 disease center visit or death during the EAP progression and clinical and follow-up period outcomes of mAb15160 Proportion of participants with To explore relationships COVID-19-related hospitalization or between mAb15160 exposure death during the EAP and follow-up and selected efficacy and period safety endpoints and/or Proportion of participants requiring biomarkers supplemental oxygen due to COVID-19 during the EAP and follow-up period Proportion of participants admitted to an intensive care unit (ICU) due to COVID-19 until the end of the follow-up period Proportion of participants requiring mechanical ventilation due to COVID-19 until the end of the follow-up period Absolute levels, change from baseline and percentage change from baseline in serum anti-SARS-CoV-2 antibodies such as anti-nucleocapsid protein IgG measured at baseline and after administration of mAb15160 over time Absolute values of serum neutralizing inhibitory dilution 50% (ID50) titers at baseline and post- mAb15160 treatment against D614G and omicron variants Change from baseline in ID50 after mAb15160 treatment against D614G and omicron variants PATIENT POPULATION Number of Subjects Approximately 5000 participants are enrolled in the study. Study Population This study enrolls participants with or without risk factors for progression to severe COVID-19, and who are RT-PCR negative at study entry. Inclusion Criteria 1. Age 12 years of age at screening. Note: participants 12 and <18 years of age must be immunocompromised and will only be enrolled where permitted by local requirements. 2. Has a SARS-CoV-2-negative RT-PCR result from a sample collected 72 hours prior to randomization or using a local RT-PCR test and sample collection following assay standards. Notes: The result should be reviewed and confirmed negative prior to dosing. Nasopharyngeal swab sample is highly preferred but other sample types (such as nasal, oropharyngeal [OP], or saliva) adhering to local standards are acceptable. 3. Is willing and able to: a. Provide informed consent signed by study participant or legally acceptable representative b. Comply with clinic visits and study-related procedures, including providing nasopharyngeal swab samples 4. Is judged by the investigator to be in stable health (including those who have a chronic stable medical condition) based on medical history, physical examination, vital sign measurements, and laboratory measurements performed at screening and/or prior to administration of study drug Exclusion Criteria 1. Has a life expectancy of less than 2 years 2. Has symptoms consistent with COVID-19 (as determined by the investigator) 3. Has a history of COVID-19 infection or has received a SARS-CoV- 2 investigational, authorized, or approved vaccine within 90 days prior to randomization 4. Planned use of any investigational, authorized, or approved vaccine for COVID-19 during the EAP 5. Prior, current, or planned use of any of the following treatments: COVID-19 convalescent plasma, other monoclonal antibodies against SARS-CoV-2 (eg, bamlanivimab and etesevimab, tixagevimab and cilgavimab, sotrovimab), or any other COVID-19 treatment (authorized, approved, or investigational) Note: prior use is defined as the past 30 days or within 5 half-lives of the treatment (whichever is longer) from screening 6. Is planned to initiate intravenous immunoglobulin (IVIG) or subcutaneous immunoglobulin (SCIG) therapy 7. Has known active symptomatic infection with influenza or other respiratory pathogen, confirmed by a diagnostic test 8. Current hospitalization or was hospitalized (ie, >24 hours) for any reason within 30 days of the screening visit 9. Has known allergy or hypersensitivity to components of the study drugs 10. Is pregnant or is breastfeeding. 11. Is a woman of childbearing potential (WOCBP).sup.1 who is unwilling to practice highly effective contraception prior to the initial dose/start of the first treatment, during the study, and for at least 6 months after the last dose. Highly effective contraceptive measures include: a. Abstinence.sup.2,3 b. Stable use of combined (estrogen and progestogen containing) hormonal contraception (oral, intravaginal, transdermal) or progestogen-only hormonal contraception (oral, injectable, implantable) associated with inhibition of ovulation initiated 2 or more menstrual cycles prior to screening c. Intrauterine device (IUD) or intrauterine hormone-releasing system (IUS) d. Bilateral tubal ligation .sup.1WOCBP are defined as women who are fertile following menarche until becoming postmenopausal, unless permanently sterile. Permanent sterilization methods include hysterectomy, bilateral salpingectomy, and bilateral oophorectomy. A postmenopausal state is defined as no menses for 12 months without an alternative medical cause. A high follicle stimulating hormone (FSH) level in the postmenopausal range may be used to confirm a postmenopausal state in women not using hormonal contraception or hormonal replacement therapy. However, in the absence of 12 months of amenorrhea, a single FSH measurement is insufficient to determine the occurrence of a postmenopausal state. The above definitions are according to the Clinical Trial Facilitation Group (CTFG) guidance. Pregnancy testing and contraception are not required for women with documented hysterectomy or tubal ligation. .sup.2Sexual abstinence is considered a highly effective method only if defined as refraining from heterosexual intercourse during the entire period of risk associated with the study drugs. The reliability of sexual abstinence needs to be evaluated in relation to the duration of the clinical trial and the preferred and usual lifestyle of the patient. Periodic abstinence (calendar, symptothermal, post-ovulation methods), withdrawal (coitus interruptus), spermicides only, and lactational amenorrhea method (LAM) are not acceptable methods 12. Is in, or is planned to enter a quarantine center 13. Is a member of the clinical site study team or their immediate family member

TABLE-US-00039 TABLE39 SequencesExcludedfromST.26-Formatted SequenceListing SEQIDNO: Sequence 13 gaggtcagt 14 EVS 33 gagggcaat 34 EGN 67 gagggcact 68 EGT 85 gaggacagt 86 EDS 105 gctgcatcc 106 AAS 125 ggtgcatcc 126 GAS 163 ggtaacagc 164 GNS 183 agtaatgat 184 SND 203 gacaatgat 204 DND 241 gatgcatcc 242 DAS 261 ggtgcaaca 262 GAT 299 agtgataat 300 SDN 319 gtcaataat 320 VNN 391 aaggcatct 392 KAS 445 gataaaaac 446 DKN 464 gaactcact 465 ELT 484 gatgtcact 485 DVT 504 gaggtcact 505 EVT 524 gagggcagt 525 EGS 541 gatgtcagt 542 DVS 577 gaaaataat 578 ENN 633 ttgggttct 634 LGS 653 ttgggatct 705 tctgcatcc 706 SAS 775 aaagacagt 776 KDS 795 ggtaacacc 796 GNT 846 aagatttct 847 KIS 915 ggtcacacc 916 GHT 932 aggaataat 933 RNN 968 tgggcatct 969 WAS 1000 ggtgcatcc 1001 GAS 1021 gctgcatcc 1022 AAS 1041 gatgtcagt 1042 DVS 1061 ggtaacagc 1062 GNS

[0339] All references cited herein are incorporated by reference to the same extent as if each individual publication, database entry (e.g., Genbank sequences or GeneID entries), patent application, or patent, was specifically and individually indicated to be incorporated by reference. This statement of incorporation by reference is intended by Applicants to relate to each and every individual publication, database entry (e.g., Genbank sequences or GeneID entries), patent application, or patent identified even if such citation is not immediately adjacent to a dedicated statement of incorporation by reference. The inclusion of dedicated statements of incorporation by reference, if any, within the specification does not in any way weaken this general statement of incorporation by reference. Citation of the references herein is not intended as an admission that the reference is pertinent prior art, nor does it constitute any admission as to the contents or date of these publications or documents