ANTIBODIES BINDING TO F-PROTEIN OF METAPNEUMOVIRUS AND USES THEREOF

Abstract

The present invention relates to antibodies, and antigen binding fragments thereof, that bind to the F-protein (fusion protein) of metapneumovims (MPV). The antibodies, and antigen binding fragments thereof, neutralize infection of MPV. The invention also relates to nucleic acids that encode, and to cells that express such antibodies and antibody fragments. In addition, the invention relates to the use of the antibodies and antibody fragments in methods for detecting and checking an MPV antigen as well as in the diagnosis, treatment and prevention of MPV infection.

Claims

1. An antibody, or an antigen-binding fragment thereof, which binds to the F-protein of metapneumovirus (MPV).

2. The antibody, or an antigen-binding fragment thereof, according to claim 1, wherein the antibody or the antigen-binding fragment thereof comprises (i) heavy chain CDR1, CDR2, and CDR3 sequences having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 7, respectively; or (ii) heavy chain CDR1, CDR2, and CDR3 sequences having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 6, and SEQ ID NO: 7, respectively; or (iii) heavy chain CDR1, CDR2, and CDR3 sequences having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 18, respectively; or (iv) heavy chain CDR1, CDR2, and CDR3 sequences having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 17, and SEQ ID NO: 18, respectively.

3. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof binds to the pre-fusion F protein of MPV.

4. The antibody, or an antigen-binding fragment thereof, according to claim 3, wherein an at least 100 fold higher concentration of the antibody, or the antigen-binding fragment thereof, is required for 50% antibody binding to post-fusion F protein of MPV than for 50% antibody binding to pre-fusion F protein of MPV.

5. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof neutralizes infection of MPV.

6. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof binds specifically to F-proteins of MPV subgroups A1, A2, B1, and B2.

7. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof neutralizes infection of MPV subgroups A1, A2, B1, and B2.

8. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises (i) heavy chain CDR1, CDR2, and CDR3 sequences having at least 80% sequence identity with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 80% sequence identity with the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 7, respectively; or (ii) heavy chain CDR1, CDR2, and CDR3 sequences having at least 80% sequence identity with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 80% sequence identity with the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 6, and SEQ ID NO: 7, respectively; or (iii) heavy chain CDR1, CDR2, and CDR3 sequences having at least 80% sequence identity with the amino acid sequences of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 80% sequence identity with the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 18, respectively; or (iv) heavy chain CDR1, CDR2, and CDR3 sequences having at least 80% sequence identity with the amino acid sequences of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 80% sequence identity with the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 17, and SEQ ID NO: 18, respectively.

9. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises (i) heavy chain CDR1, CDR2, and CDR3 sequences having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 7, respectively; or (ii) heavy chain CDR1, CDR2, and CDR3 sequences having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, and SEQ ID NO: 3, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 4, SEQ ID NO: 6, and SEQ ID NO: 7, respectively; or (iii) heavy chain CDR1, CDR2, and CDR3 sequences having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 18, respectively; or (iv) heavy chain CDR1, CDR2, and CDR3 sequences having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 12, SEQ ID NO: 13, and SEQ ID NO: 14, respectively, and light chain CDR1, CDR2, and CDR3 sequences having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 15, SEQ ID NO: 17, and SEQ ID NO: 18, respectively.

10. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 1; a heavy chain CDR2 sequence according to SEQ ID NO: 2; a heavy chain CDR3 sequence having at least 90% sequence identity with the amino acid sequences of SEQ ID NO: 3; a light chain CDR1 sequence according to SEQ ID NO: 4; a light chain CDR2 sequence according to SEQ ID NO: 5 or 6; and a light chain CDR3 sequence according to SEQ ID NO: 7.

11. The antibody, or an antigen-binding fragment thereof, according to claim 10, wherein the C-terminal Asp residue in SEQ ID NO: 3 is substituted; optionally with another polar amino acid.

12. The antibody, or an antigen-binding fragment thereof, according to claim 10 or 11, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain CDR3 sequence according to SEQ ID NO: 3 or 10.

13. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 1; a heavy chain CDR2 sequence according to SEQ ID NO: 2; a heavy chain CDR3 sequence according to SEQ ID NO: 3; a light chain CDR1 sequence according to SEQ ID NO: 4; a light chain CDR2 sequence according to SEQ ID NO: 5 or 6; and a light chain CDR3 sequence according to SEQ ID NO: 7.

14. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 1; a heavy chain CDR2 sequence according to SEQ ID NO: 2; a heavy chain CDR3 sequence according to SEQ ID NO: 10; a light chain CDR1 sequence according to SEQ ID NO: 4; a light chain CDR2 sequence according to SEQ ID NO: 5 or 6; and a light chain CDR3 sequence according to SEQ ID NO: 7.

15. The antibody, or an antigen-binding fragment thereof, according to any one of claims 1 to 9, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence having at least 70% sequence identity with the amino acid sequences of SEQ ID NO: 12; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

16. The antibody, or an antigen-binding fragment thereof, according to claim 15, wherein one or more of the heavy chain variable region amino acid residues N34, S36 and C38 (corresponding to N6, S8 and C10, respectively, in SEQ ID NO: 12) is/are substituted.

17. The antibody, or an antigen-binding fragment thereof, according to claim 16, wherein N34 (corresponding to N6 in SEQ ID NO: 12) is substituted with Gln (Q) or Ser (S); S36 (corresponding to S8 in SEQ ID NO: 12) is substituted with Ala (A); and/or C38 (corresponding to C10 in SEQ ID NO: 12) is substituted with Ser (S), Ala (A) or Tyr (Y).

18. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 17, wherein the heavy chain CDR1 sequence differs in a single or exactly two amino acid substitution(s) from SEQ ID NO: 12.

19. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 18, wherein the antibody, or an antigen-binding fragment thereof, comprises a heavy chain CDR1 sequence according to any one of SEQ ID NOs 12, 21, 23, 25, 27, 29, 31 and 33.

20. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 19, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 12; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

21. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 19, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 21; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

22. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 19, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 23; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

23. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 19, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 25; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

24. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 19, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 27; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

25. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 19, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 29; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

26. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 19, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 31; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

27. The antibody, or an antigen-binding fragment thereof, according to any one of claims 15 to 19, wherein the antibody or the antigen-binding fragment thereof comprises: a heavy chain CDR1 sequence according to SEQ ID NO: 33; a heavy chain CDR2 sequence according to SEQ ID NO: 13; a heavy chain CDR3 sequence according to SEQ ID NO: 14; a light chain CDR1 sequence according to SEQ ID NO: 15; a light chain CDR2 sequence according to SEQ ID NO: 16 or 17; and a light chain CDR3 sequence according to SEQ ID NO: 18.

28. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises (i) a heavy chain variable region comprising an amino acid sequence having at least 70% identity to SEQ ID NO: 8 and a light chain variable region comprising the amino acid sequence having at least 70% identity to SEQ ID NO: 9; or (ii) a heavy chain variable region comprising an amino acid sequence having at least 70% identity to SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence having at least 70% identity to SEQ ID NO: 20.

29. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises (i) a heavy chain variable region comprising an amino acid sequence having at least 75% identity to SEQ ID NO: 8 and a light chain variable region comprising the amino acid sequence having at least 75% identity to SEQ ID NO: 9; or (ii) a heavy chain variable region comprising an amino acid sequence having at least 75% identity to SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence having at least 75% identity to SEQ ID NO: 20.

30. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises (i) a heavy chain variable region comprising an amino acid sequence having at least 80% identity to SEQ ID NO: 8 and a light chain variable region comprising the amino acid sequence having at least 80% identity to SEQ ID NO: 9; or (ii) a heavy chain variable region comprising an amino acid sequence having at least 80% identity to SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence having at least 80% identity to SEQ ID NO: 20.

31. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises (i) a heavy chain variable region comprising an amino acid sequence having at least 85% identity to SEQ ID NO: 8 and a light chain variable region comprising the amino acid sequence having at least 85% identity to SEQ ID NO: 9; or (ii) a heavy chain variable region comprising an amino acid sequence having at least 85% identity to SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence having at least 85% identity to SEQ ID NO: 20.

32. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises (i) a heavy chain variable region comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 8 and a light chain variable region comprising the amino acid sequence having at least 90% identity to SEQ ID NO: 9; or (ii) a heavy chain variable region comprising an amino acid sequence having at least 90% identity to SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence having at least 90% identity to SEQ ID NO: 20.

33. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises (i) a heavy chain variable region comprising an amino acid sequence having at least 95% identity to SEQ ID NO: 8 and a light chain variable region comprising the amino acid sequence having at least 95% identity to SEQ ID NO: 9; or (ii) a heavy chain variable region comprising an amino acid sequence having at least 95% identity to SEQ ID NO: 19 and a light chain variable region comprising the amino acid sequence having at least 95% identity to SEQ ID NO: 20.

34. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 8 and a light chain variable region according to SEQ ID NO: 9.

35. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 11 and a light chain variable region according to SEQ ID NO: 9.

36. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 19 and a light chain variable region according to SEQ ID NO: 20.

37. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 22 and a light chain variable region according to SEQ ID NO: 20.

38. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 24 and a light chain variable region according to SEQ ID NO: 20.

39. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 26 and a light chain variable region according to SEQ ID NO: 20.

40. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 28 and a light chain variable region according to SEQ ID NO: 20.

41. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 30 and a light chain variable region according to SEQ ID NO: 20.

42. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 32 and a light chain variable region according to SEQ ID NO: 20.

43. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof comprises a heavy chain variable region comprising an amino acid sequence according to SEQ ID NO: 34 and a light chain variable region according to SEQ ID NO: 20.

44. The antibody, or an antigen-binding fragment thereof, according to any one of the previous claims, wherein the antibody or the antigen-binding fragment thereof is a human antibody.

45. The antibody, or an antigen-binding fragment thereof, of any one of the previous claims, wherein the antibody, or an antigen-binding fragment thereof, is a monoclonal antibody.

46. The antibody of any one of the previous claims, wherein the antibody comprises an Fc moiety.

47. The antibody of any one of the previous claims, wherein the antibody is of the IgG type.

48. The antibody of claim 47, wherein the antibody is of the IgG1 type.

49. The antibody, or an antigen-binding fragment thereof, of any one of the previous claims, wherein the antibody, or the antigen-binding fragment thereof, is purified.

50. The antibody, or an antigen-binding fragment thereof, of any one of the previous claims, wherein the antibody, or the antigen-binding fragment thereof, is a single-chain antibody.

51. The antibody, or an antigen-binding fragment thereof, of any one of the previous claims, wherein the antibody, or the antigen-binding fragment thereof, is selected from Fab, Fab, F(ab)2, Fv or scFv.

52. The antibody, or an antigen-binding fragment thereof, of any one of the previous claims for use as a medicament.

53. The antibody, or an antigen-binding fragment thereof, for use according to claim 52 in prophylaxis or treatment of MPV infection.

54. A nucleic acid molecule comprising a polynucleotide encoding the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51.

55. The nucleic acid molecule of claim 54, wherein the polynucleotide encoding the antibody, or an antigen-binding fragment thereof, is codon-optimized.

56. The nucleic acid molecule of claim 54 or 55 comprising a nucleic acid sequence as set forth in any one of SEQ ID NOs 38-55; or a sequence variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity.

57. A combination of a first and a second nucleic acid molecule, wherein the first nucleic acid molecule comprises a polynucleotide encoding the heavy chain of the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51; and the second nucleic acid molecule comprises a polynucleotide encoding the corresponding light chain of the same antibody, or the same antigen-binding fragment thereof.

58. The combination of nucleic acid molecules of claim 57, wherein one or both of the polynucleotides encoding the heavy and/or light chain(s) of the antibody, or an antigen-binding fragment thereof, is/are codon-optimized.

59. The combination of nucleic acid molecules of claim 57 or 58 comprising a nucleic acid sequence as set forth in any one of SEQ ID NOs 38-55; or a sequence variant thereof having at least 70%, at least 75%, at least 80%, at least 85%, at least 88%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity.

60. A combination of a first and a second nucleic acid molecule, wherein (i) the first nucleic acid molecule comprises a polynucleotide encoding the heavy chain of an antibody, or an antigen-binding fragment thereof, the polynucleotide comprising: (a) nucleotide sequences according to SEQ ID NOs 38, 39 and 40; or (b) nucleotide sequences according to SEQ ID NOs 47, 48 and 49; and (ii) the second nucleic acid molecule comprises a polynucleotide encoding the light chain of an antibody, or an antigen-binding fragment thereof, the polynucleotide comprising: (c) nucleotide sequences according to SEQ ID NOs 41, 42 (or 43) and 44; or (d) nucleotide sequences according to SEQ ID NOs 50, 51 (or 52) and

53.

61. A vector comprising the nucleic acid molecule of any one of claims 54 to 56 or the combination of nucleic acid molecules of any one of claims 57 to 60.

62. A combination of a first and a second vector, wherein the first vector comprises a first nucleic acid molecule as defined in any one of claims 57 to 60 and the second vector comprises the corresponding second nucleic acid molecule as defined in any one of claims 57 to 60.

63. A cell expressing the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51, or comprising the vector of claim 61 or the combination of vectors of claim 62.

64. A pharmaceutical composition comprising the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51, the nucleic acid of any one of claims 54 to 56, the combination of nucleic acids of any one of claims 57 to 60, the vector of claim 61, the combination of vectors of claim 62 or the cell of claim 63, and, optionally, a pharmaceutically acceptable excipient, diluent or carrier.

65. The antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51, the nucleic acid of any one of claims 54 to 56, the combination of nucleic acids of any one of claims 57 to 60, the vector of claim 61, the combination of vectors of claim 62, the cell of claim 63, or the pharmaceutical composition of claim 64 for use as a medicament; optionally in the prophylaxis or treatment of MPV infection.

66. Use of the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51 in (in-vitro) diagnosis of MPV infection.

67. Use of the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51 in a method for detecting MPV antigens.

68. Use of the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51 for monitoring the quality of anti-MPV vaccines by checking the antigen of said vaccine.

69. The use according to claim 68, wherein the conformation of the antigen, or an epitope thereof, of said vaccine is checked.

70. Use of the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51, the nucleic acid of any one of claims 54 to 56, the combination of nucleic acids of any one of claims 57 to 60, the vector of claim 61, the combination of vectors of claim 62, the cell of claim 63, or the pharmaceutical composition of claim 64 in the manufacture of a medicament for prophylaxis, treatment or attenuation of MPV infection.

71. A method of reducing MPV infection, or lowering the risk of MPV infection, comprising: administering to a subject in need thereof, a therapeutically effective amount of the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51, the nucleic acid of any one of claims 54 to 56, the combination of nucleic acids of any one of claims 57 to 60, the vector of claim 61, the combination of vectors of claim 62, the cell of claim 63, or the pharmaceutical composition of claim 64.

72. A method for testing an anti-MPV vaccine, wherein the vaccine is contacted with the antibody, or an antigen-binding fragment thereof, of any one of claims 1 to 51 and, optionally, the presence of antibody/antigen complexes is determined.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0339] In the following a brief description of the appended figures will be given. The figures are intended to illustrate the present invention in more detail. However, they are not intended to limit the subject matter of the invention in any way.

[0340] FIG. 1 shows for Example 1 the binding properties of MPE33 and MPF5 antibodies to the hMPV F proteins in a pre-fusion and post-fusion conformation, in comparison to the reference antibody MPE8.

[0341] FIG. 2 shows for Example 2 the neutralization of four distinct hMPV strains A1/6621 (MPV A1), A2/VR8938 (MPV A2), B1/VR4702 (MPV B1), and B2/3817 (MPV B2) with antibodies MPE33 and MPF5 in comparison to the reference antibody MPE8v3.

[0342] FIG. 3 summarizes for Example 2 the EC50 required for neutralization of the four distinct hMPV strains A1/6621 (MPV A1), A2/VR8938 (MPV A2), B1/VR4702 (MPV B1), and B2/3817 (MPV B2) with antibodies MPE33 and MPF5 in comparison to the reference antibody MPE8v3.

[0343] FIG. 4 shows for Example 3 the binding affinities and EC50 values of antibodies MPF5_VH117D (MPF5), MPF5_VH117H, MPE33 and comparative antibody MPE8v3 for MPV pre-fusion F protein.

[0344] FIG. 5 shows the VH and VL (VK) sequences of antibodies MPF5_VH117D (MPF5) and MPF5_VH117H. The square indicates the position of the substitution at amino acid position VH117.

[0345] FIG. 6 shows for Example 4 the binding affinities and EC50 values of antibody MPE33 and its variants MPE33_S36A, MPE33_N34Q, MPE33_N34S, MPE33_C38S, MPE33_C38A, MPE33_C38Y and MPE33_N34S_C38Y for MPV pre-fusion F protein.

[0346] FIG. 7 shows the VH and VL (VK) sequences of antibody MPE33. The squares indicate the positions of the substitutions at amino acid position N34, S36 and C38.

[0347] FIG. 8 shows for Example 5 the binding of antibodies MPE33, MPE33_S36A, MPE33_N34Q, MPE33_N34S, MPE33_C38S, MPE33_C38A, MPE33_C38Y, MPE33_N34S_C38Y, MPF5_VH117D (MPF5), MPF5_VH117H and comparative antibody MPE8 to cell-associated F-antigen of hMPV strains MPV_NL/1/99_F0-TM (upper line) and HMPV_Yokohama/JPN(P8527)2016 (middle line). In the lower line, mock transfection (control) is shown.

[0348] FIG. 9 shows for Example 6 the results of a competition study of antibodies MPF5, MPE33 and MPE8 for binding to MPV F-protein. The data show competition between the same antibodies (MPF5 vs. MPF5; MPE33 vs. MPE33; MPE8 vs. MPE8), as expected. In addition, panels MPF5 vs. MPE8 and MPE8 vs. MPF5 demonstrate competition between MPF5 and MPE8. No competition, however, could be found between MPE33 and MPE8, or MPE33 and MPF5.

EXAMPLES

[0349] In the following, particular examples illustrating various embodiments and aspects of the invention are presented. However, the present invention shall not to be limited in scope by the specific embodiments described herein. The following preparations and examples are given to enable those skilled in the art to more clearly understand and to practice the present invention. The present invention, however, is not limited in scope by the exemplified embodiments, which are intended as illustrations of single aspects of the invention only, and methods which are functionally equivalent are within the scope of the invention. Indeed, various modifications of the invention in addition to those described herein will become readily apparent to those skilled in the art from the foregoing description, accompanying figures and the examples below. All such modifications fall within the scope of the appended claims.

Example 1: Identification and Characterization of Human Monoclonal Antibodies MPF5 and MPE33

[0350] Human monoclonal antibodies MPF5 (also referred to as MPF5_VH117D) and MPE33 against MPV were isolated (cf. Traggiai E. et al., 2004, Nat Med 10(8): 871-5) from human patients. The antibodies were characterized by determining the nucleotide and amino acid sequences of its variable regions (MPF5 VH: SEQ ID NO: 8, MPF5 VL: SEQ ID NO: 9; MPE33 VH: SEQ ID NO: 19, MPE33 VL: SEQ ID NO: 20) and the complementarity determining regions (CRDs) therein (MPF5: SEQ ID NOs 1-5 and 7, or 1-4, 6 and 7, respectively; MPE33: SEQ ID NOs 12-16 and 18, or 12-15, 17 and 18, respectively). The VH and VL genes of MPF5 and MPE33 were cloned into IgG1 expression vectors and recombinant antibodies were produced by transient transfection of 293 Freestyle cells (293F). Supernatants from transfected cells were collected and IgG were affinity purified by Protein A chromatography. Accordingly, MPF5 and MPE33 are IgG1-type fully human monoclonal antibodies with the CDR, VH and VL sequences as described herein.

[0351] In order to test the binding of MPF5 and MPE33 to hMPV F protein, an ELISA assay was performed essentially as described in WO 2016/103238 A1 for testing the binding affinity of the antibodies to hMPV F protein in pre- vs. post-fusion conformation. For comparison, prior art antibody MPE8 (Corti et at, 2013, Cross-neutralization of four paramyxoviruses by a human monoclonal antibody. Nature 501: 439-443) was also tested in this experiment.

[0352] Briefly, Maxisorp ELISA plates were coated overnight with both conformationally stabilized pre and post fusion F protein antigens (F protein from CAN97-83 MPV strain, 1 g/ml, 25 l/well in PBS pH7). After 3 washes with PBS/Tween 0.01% (PBST), test antibodies were added starting at 10 g/ml, titrated down 3-fold by 11 points, and incubated 2 hrs at room temperature. Plates were then washed 4 times in PBST and dispensed with alkaline phosphatase-labeled goat anti-human IgG polyclonal antibody (SouthernBiotech, 2 g/ml, 25 l/well) and further incubated 1 hr at RT. Following 4 washes with PBST, plates were developed by adding 50 l/well of AP substrate (pNPP, Sigma) in carbonate buffer and read at 405 nm after 45 min.

[0353] Results are shown in FIG. 1. All antibodies tested (MPF5, MPE33 and MPE8) showed high binding affinity to MPV pre-fusion F protein, but not to MPV post-fusion F protein. Accordingly, antibodies MPF5, MPE33 and MPE8 are specific for MPV pre-fusion F protein. Antibodies MPF5 and MPE33 of the present invention showed even lower EC50 values and, thus, higher binding affinity to MPV pre-fusion F protein as compared to prior art antibody MPE8.

Example 2: MPF5 and MPE33 Effectively Neutralize Various Strains of MPV

[0354] Next, neutralization of various distinct strains of MPV was assessed with the antibodies MPF5 and MPE33 of the present invention as well with comparative antibody MPE8v3, which differs from control antibody MPE8 used in Example 1 in that it comprises a N113S mutation in the heavy chain variable region to remove a glycosylation site.

[0355] Briefly, culture supernatants with the antibodies were analyzed using a microneutralization assay based on infection of LLC-MK2 cells by hMPV strains A1/6621, A2/VR8938, B1/VR4702, and B2/3817. Neat supernatants were incubated with 0.239*10.sup.6 TCID50/ml (A1/6621), 0.0959*10.sup.6 TCID50/ml (A2/VR8938), 0.33*10.sup.6 TCID50/ml (B1/VR4702) and 0.0959*10.sup.6 TCID50/ml (B2/3817), respectively, of viruses for 1 h at room temperature before addition of LLC-MK2 target cells which were incubated for 14 days, respectively. Viable cells were detected using the WST-1 reagent (Roche). The EC50 was determined using the microneutralization assay described above with 100 TCID.sub.50 of virus and viral infection was measured on day 6 or 7 by indirect immunofluorescence using an automated Pathway 855 analyser (BD). EC50 values were calculated by interpolation ,of neutralization curves fitted with a 4-parameter nonlinear regression with a variable slope.

[0356] Results are shown in FIGS. 2 and 3. All antibodies effectively neutralized the four different MPV strains tested. Specifically, MPE33 and MPF5 showed a significantly better neutralization potency against MPV B2 strain, when compared to the reference mAb MPE8v3.

Example 3: CDRH3 Mutant of MPF5 Shows Similarly High Binding Affinity to MPV Pre-Fusion F Protein

[0357] Next, variant antibody MPF5_VH117H of MPF5 was generated, which differs from MPF5 in that its CDRH3 sequence is according to SEQ ID NO: 10 and its VH sequence is according to SEQ ID NO: 11. The differences in the heavy chain sequence and CDRH3 of MPF5 are illustrated in FIG. 5.

[0358] The binding affinities of antibodies MPF5_VH117D (MPF5), MPF5_VH117H, MPE8v3 and MPE33 for MPV pre-fusion F protein were tested in an ELISA assay as described in Example 1.

[0359] Results are shown in FIG. 4. While all tested antibodies specifically bound to MPV pre-fusion F protein, the binding affinities of the antibodies MPF5_VH117D (MPF5), MPF5_VH117H and MPE33 of the present invention were considerably higher than that of comparative antibody MPE8v3. Binding affinities of antibodies MPF5_VH117D (MPF5) and MPF5_VH117H were similarly high, indicating that the CDRH3 mutation did not impair binding affinity of MPF5 to MPV pre-fusion F protein.

Example 4: CDRH1 Mutants of MPE33 Show Similarly High Binding Affinity to MPV Pre-Fusion F Protein

[0360] Next, the following variant antibodies of MPE33 were generated, which differ from MPE33 in the indicated CDRH1 sequence only:

TABLE-US-00003 TABLE 3 CDRH1 VH Antibody (SEQ ID NO) (SEQ ID NO) MPE33 12 19 MPE33_S36A 21 22 MPE33_N34Q 23 24 MPE33_N34S 25 26 MPE33_C38S 27 28 MPE33_C38A 29 30 MPE33_C38Y 31 32 MPE33_N34S_C38Y 33 34

[0361] The positions of the mutated amino acids in the heavy chain sequence and CDRH3 of MPE33 are illustrated in FIG. 7.

[0362] The binding affinities of antibodies MPE33 and its variant antibodies described above for MPV pre-fusion F protein were tested in an ELISA assay as described in Example 1.

[0363] Results are shown in FIG. 6. All tested antibodies MPE33, MPE33_S36A, MPE33_N34Q, MPE33_N34S, MPE33_C38S, MPE33_C38A, MPE33_C38Y and MPE33_N34S_C38Y specifically bound to MPV pre-fusion F protein with similarly high binding affinities. The binding affinities of all variant antibodies were even slightly higher than the binding affinity of MPE33 for MPV pre-fusion F protein, indicating that the various CDRH1 mutations did not impair binding affinity of MPE33 to MPV pre-fusion F protein.

Example 5: Binding to Cell-Associated F Antigen

[0364] Next, binding of all exemplified antibodies of the present invention as described in the above examples as well as of comparative antibody MPE8 was tested.

[0365] To this end, Expi293 cells were transfected with MPV F-protein (MPV_NL/1/99_F0-TM (AY304361) and HMPV_Yokohama/JPN(P8527)2016). HMPV_Yokohama/JPN(P8527)2016 carries D280N mutation. Briefly, 10 g of plasmid DNA were diluted in 0.5 mL of Opti-Mem I medium (Gibco, cat. #31985-047), added to 0.5 mL Opti-Mem containing 30 l PEI Max transfection reagent (40 kD, cat. # POL24765-1, Polysciences) and incubated 20 min at room temperature (RT). Transfection mix was then added to a culture flask containing 80 ml of expression medium (GIBCO, cat# A14351-02) with growing Expi293 cells (310.sup.6 cells/mL) and further incubated 3 days at 37 C. under agitation. Cells were then harvested, fixed in 4% formaldehyde 20 min on ice, permeabilized with 0.5% Saponin, 1% FBS in PBS 20 min on ice and stained with antibodies MPE33, MPE33_S36A, MPE33_N34Q, MPE33_N34S, MPE33_C38S, MPE33_C38A, MPE33_C38Y and MPE33_N34S_C38Y, MPF5_VH117D (MPF5), MPF5_VH117H and comparative antibody MPE8 (5 g/ml, 60 min at 4 C. in permeabilization buffer). Binding was revealed by staining cells with AF 647 Goat Anti-Human IgG, Fc Fragment Specific secondary antibody (Jackson, 109-606-098, 1 g/ml 30 min on ice) and by acquiring cells with a flow cytometer.

[0366] Results are shown in FIG. 8 with MPV_NL/1/99_F0-TM in the upper line, HMPV_Yokohama/JPN(P8527)2016 in the middle line and mock transfection in the lower line. These data show that MPE33 and MPF5 as well as all their variants bind equally well to the F protein of both B1 (NL1/1/99) and B2 (Yokohama) hMPV strains, while the B2 strain (which carries the D280N mutation) results in a viral escape variant for the MPE8 antibody.

Example 6: Competition Study of Antibodies MPE33, MPF5 and MPE8

[0367] To identify whether antibodies MPE33 and MPF5 bind to the same or distinct epitopes on MPV F-protein as comparative antibody MPE8, a binding/competition study was performed.

[0368] To this end, competition of antibodies MPE33, MPF5 and MPE8 was assessed by bio-layer interferometry using OCTET RED96 (ForteBio). Briefly, APS sensors were loaded/coated with MPV F-protein at 5 g/ml in PBS for 10 minutes. Successively sensors were blocked using BSA 1 mg/ml in PBS (blocking buffer) for 5 minutes. Association of the mAbs was performed by moving the sensor into two consecutive wells (7 minutes each) containing the first and second mAb at 30 ug/ml in blocking buffer respectively. All steps were performed at 30 C. under constant mixing at 1000 rpm.

[0369] Results are shown in FIG. 9. The data demonstrate that MPF5 competes with MPE8, indicating that both antibodies bind to the same or an overlapping epitope on MPV F-protein. MPE33, however, competes neither with MPF5 nor with MPE8, indicating that MPE33 binds to a distinct epitope on MPV F-protein as compared to MPF5 and MPE8.

TABLE-US-00004 TABLEOFSEQUENCESANDSEQIDNUMBERS(SEQUENCELISTING): SEQIDNO Sequence Remarks Aminoacidsequences MPF5_VH117D SEQIDNO:1 SVSFNDYY CDRH1 SEQIDNO:2 IGHGGEH CDRH2 SEQIDNO:3 ARGIGWLPPPD CDRH3 SEQIDNO:4 QSVLFSSNNENY CDRL1 SEQIDNO:5 WAS CDRL2 SEQIDNO:6 LIYWASTRE CDRL2long SEQIDNO:7 QQFYSPPWT CDRL3 SEQIDNO:8 QVQLQQWGAGLLKPSETLSLTCVGNSVSFNDY VH YWSWIRQSPGKGLEWIGEIGHGGEHNYNPSLN GRVTMSVDTSNNHFSLLLSSVTAADTAMYYCA RGIGWLPPPDWGQGTLVTVSS SEQIDNO:9 DIVMTQSPDSLAVSLGERATINCKSSQSVLFSS VL NNENYLAWFQQKPGQPPKLLIYWASTRESGVPD RFSGSGSGTDFTLTITSLQAEDVAVYYCQQFYS PPWTFGQGTKVEIK MPF5_VH117H SEQIDNO:10 ARGIGWLPPPH CDRH3 SEQIDNO:11 QVQLQQWGAGLLKPSETLSLTCVGNSVSFNDY VH YWSWIRQSPGKGLEWIGEIGHGGEHNYNPSLN GRVTMSVDTSNNHFSLLLSSVTAADTAMYYCA RGIGWLPPPHWGQGTLVTVSS MPE33 SEQIDNO:12 GDSISNGSYC CDRH1 SEQIDNO:13 TYPIGNT CDRH2 SEQIDNO:14 AREARIFEGYYYYYYGLDV CDRH3 SEQIDNO:15 QTISSY CDRL1 SEQIDNO:16 DVS CDRL2 SEQIDNO:17 LISDVSKRA CDRL2long SEQIDNO:18 HQRSNWDT CDRL3 SEQIDNO:19 QVQLQESGPGLVKPSQTLSLTCTVSGDSISNGSY VH CWNWVRQPAGKGLEWIGRIYPIGNTNYNPSLKSR VTISLDTSKNQFSLKLASVTAADTAVYYCAREAR IFEGYYYYYYGLDVWGQGTTVTVSS SEQIDNO:20 EIVLTQSPATLSLSPGERATLSCRASQTISSYLAW VL YQQKPGQAPRLLISDVSKRATGIPARFSGSGSGTD FTLTISSLEPEDFAVYYCHQRSNWDTFGQGTKLE IK MPE33_S36A SEQIDNO:21 GDSISNGAYC CDRH1 SEQIDNO:22 QVQLQESGPGLVKPSQTLSLTCTVSGDSISNGA VH YCWNWVRQPAGKGLEWIGRIYPIGNTNYNPSLK SRVTISLDTSKNQFSLKLASVTAADTAVYYCAR EARIFEGYYYYYYGLDVWGQGTTVTVSS MPE33_N34Q SEQIDNO:23 GDSISQGSYC CDRH1 SEQIDNO:24 QVQLQESGPGLVKPSQTLSLTCTVSGDSISQGS VH YCWNWVRQPAGKGLEWIGRIYPIGNTNYNPSLK SRVTISLDTSKNQFSLKLASVTAADTAVYYCAR EARIFEGYYYYYYGLDVWGQGTTVTVSS MPE33_N34S SEQIDNO:25 GDSISSGSYC CDRH1 SEQIDNO:26 QVQLQESGPGLVKPSQTLSLTCTVSGDSISSGS VH YCWNWVRQPAGKGLEWIGRIYPIGNTNYNPSLK SRVTISLDTSKNQFSLKLASVTAADTAVYYCAR EARIFEGYYYYYYGLDVWGQGTTVTVSS MPE33_C38S SEQIDNO:27 GDSISNGSYS CDRH1 SEQIDNO:28 QVQLQESGPGLVKPSQTLSLTCTVSGDSISNGS VH YSWNWVRQPAGKGLEWIGRIYPIGNTNYNPSLK SRVTISLDTSKNQFSLKLASVTAADTAVYYCAR EARIFEGYYYYYYGLDVWGOGTTVTVSS MPE33_C38A SEQIDNO:29 GDSISNGSYA CDRH1 SEQIDNO:30 QVQLQESGPGLVKPSQTLSLTCTVSGDSISNGS VH YAWNWVRQPAGKGLEWIGRIYPIGNTNYNPSLK SRVTISLDTSKNQFSLKLASVTAADTAVYYCAR EARIFEGYYYYYYGLDVWGQGTTVTVSS MPE33_C38Y SEQIDNO:31 GDSISNGSYY CDRH1 SEQIDNO:32 QVQLQESGPGLVKPSQTLSLTCTVSGDSISNGS VH YYWNWVRQPAGKGLEWIGRIYPIGNTNYNPSLK SRVTISLDTSKNQFSLKLASVTAADTAVYYCAR EARIFEGYYYYYYGLDVWGQGTTVTVSS MPE33_N34S_C38Y SEQIDNO:33 GDSISSGSYY CDRH1 SEQIDNO:34 QVQLQESGPGLVKPSQTLSLTCTVSGDSISSGS VH YYWNWVRQPAGKGLEWIGRIYPIGNTNYNPSLK SRVTISLDTSKNQFSLKLASVTAADTAVYYCAR EARIFEGYYYYYYGLDVWGQGTTVTVSS Constantregions SEQIDNO:35 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE heavychain PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVV TVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMIS RTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNA KTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPP SRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQ PENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQ GNVFSCSVLHEALHSHYTQKSLSLSPGK SEQIDNO:36 GQPKAAPSVTLFPPSSEELQANKATLVCLISDF lambdalightchain YPGAVTVAWKADSSPVKAGVETTTPSKQSNNKY AASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKT VAPTECS SEQIDNO:37 RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFY kappalightchain PREAKVQWKVDNALQSGNSQESVTEQDSKDSTY SLSSTLTLSKADYEKHKVYACEVTHQGLSSPVT KSFNRGEC Nucleicacidsequences MPF5_VH117D SEQIDNO:38 AGTGTGTCCTTCAATGATTACTAC CDRH1 SEQIDNO:39 ATCGGTCACGGTGGAGAACAC CDRH2 SEQIDNO:40 GCGAGAGGTATTGGCTGGCTCCCCCCTCCG CDRH3 GAC SEQIDNO:41 CAGAGTGTTTTATTCAGCTCCAACAATGAGAA CDRL1 CTAC SEQIDNO:42 TGGGCATCT CDRL2 SEQIDNO:43 CTCATTTACTGGGCATCTACCCGGGAA CDRL2long SEQIDNO:44 CAGCAATTTTATAGTCCTCCGTGGACG CDRL3 SEQIDNO:45 CAGGTGCAGCTTCAGCAGTGGGGCGCAGGA VH CTGTTGAAGCCTTCGGAGACCCTGTCCCTCA CCTGCGTTGGAAATAGTGTGTCCTTCAATGAT TACTACTGGAGCTGGATCCGCCAGTCCCCAG GGAAGGGGCTGGAGTGGATTGGGGAAATC GGTCACGGTGGAGAACACAACTACAACCCGT CCCTGAACGGTCGAGTCACCATGTCGGTGGAC ACGTCCAACAACCATTTCTCCCTACTTCTAT CTTCTGTGACCGCCGCGGACACGGCTATGTA TTACTGTGCGAGAGGTATTGGCTGGCTCCCC CCTCCGGACTGGGGCCAGGGAACCCTGGTC ACCGTCTCATCAG SEQIDNO:46 GACATCGTCATGACTCAGTCTCCAGACTCCC VL TGGCTGTGTCTCTGGGCGAGAGGGCCACCAT CAACTGCAAGTCCAGCCAGAGTGTTTTATTCA GCTCCAACAATGAGAACTACTTAGCTTGGTTC CAGCAGAAACCAGGACAGCCTCCTAAGCTAC TCATTTACTGGGCATCTACCCGGGAATCCGG GGTCCCTGACCGATTCAGTGGCAGCGGGTC TGGGACAGATTTCACTCTCACCATCACCAGC CTGCAGGCTGAAGATGTGGCAGTTTATTACT GTCAGCAATTTTATAGTCCTCCGTGGACGTTC GGCCAAGGGACCAAGGTGGAAATCAAAC MPE33 SEQIDNO:47 GGTGACTCCATCAGCAACGGTAGTTACTGC CDRH1 SEQIDNO:48 ATCTATCCCATTGGAAACACC CDRH2 SEQIDNO:49 GCGAGAGAGGCGAGGATCTTTGAAGGCTAT CDRH3 TACTACTACTATTACGGTTTGGACGTC SEQIDNO:50 CAGACTATAAGTAGTTAC CDRL1 SEQIDNO:51 GATGTATCC CDRL2 SEQIDNO:52 CTCATCTCTGATGTATCCAAAAGGGCC CDRL2long SEQIDNO:53 CACCAACGTAGCAACTGGGACACT CDRL3 SEQIDNO:54 CAGGTGCAGCTGCAGGAGTCGGGCCCAGG VH ACTGGTGAAGCCTTCACAGACCCTGTCCCTC ACCTGCACTGTCTCTGGTGACTCCATCAGCA ACGGTAGTTACTGCTGGAATTGGGTCCGGCA GCCCGCCGGGAAGGGACTGGAGTGGATTG GGCGTATCTATCCCATTGGAAACACCAACTA CAACCCCTCCCTCAAGAGTCGAGTCACCATA TCACTAGACACGTCCAAGAACCAGTTCTCCC TGAAGCTGGCTTCTGTGACCGCCGCAGACAC GGCCGTCTACTACTGTGCGAGAGAGGCGAGG ATCTTTGAAGGCTATTACTACTACTATTACG GTTTGGACGTCTGGGGCCAAGGGACCACGG TCACCGTCTCCTCAG SEQIDNO:55 GAAATTGTGTTGACACAGTCTCCAGCCACCC VL TGTCTTTGTCTCCAGGGGAAAGAGCCACCCT CTCCTGCAGGGCCAGTCAGACTATAAGTAGT TACTTAGCCTGGTACCAACAGAAACCTGGCC AGGCTCCCAGGCTCCTCATCTCTGATGTATC CAAAAGGGCCACTGGCATCCCAGCCAGGTTC AGTGGCAGTGGGTCTGGGACAGACTTCACT CTCACCATCAGCAGCCTAGAGCCTGAAGATT TTGCAGTTTATTACTGTCACCAACGTAGCAA CTGGGACACTTTTGGCCAGGGGACTAAGCTG GAGATCAAAC