Anti-OSPA antibodies and methods of use

• US 10,457,721 B2
• Filed: 08/07/2015
• Issued: 10/29/2019
• Est. Priority Date: 08/11/2014
• Status: Active Grant

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Wang, Yang, Thomas, Jr., William D., Boatright, Naomi K., Klempner, Mark S. J.
• Primary Examiner(s): Wu, Julie
• Application Number:
  US15/501,469
• Publication Number:
  US 20170226193A1
• Time in Patent Office: 1,544 Days
• Field of Search: None
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61P 31/04 Antibacterial agents
• CPC Class Codes: C07K 16/1207 from Spirochaetales (O), e.g. Treponema, Leptospira
• CPC Class Codes: C07K 2317/21 from primates, e.g. man
• CPC Class Codes: C07K 2317/52 Constant or Fc region; Isotype
• CPC Class Codes: C07K 2317/565 Complementarity determining region [CDR]
• CPC Class Codes: C07K 2317/72 Increased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/73 Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C12P 21/005 Glycopeptides, glycoproteins
• CPC Class Codes: G01N 33/56911 Bacteria
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The invention provides anti-OspA antibodies and methods of using the same.

Claims

1. An isolated antibody that specifically binds to outer surface protein A (OspA), wherein the antibody comprises the following six CDRs:

(a) a CDR-H1 comprising the amino acid sequence of GYSFTSYWIG (SEQ ID NO;

     10);

(b) a CDR-H2 comprising the amino acid sequence of FIYPGDSDTRYSPSFQG (SEQ ID NO;

     11);

(c) a CDR-H3 comprising the amino acid sequence of ARGILRYFDWFLDY (SEQ ID NO;

     4);

(d) a CDR-L1 comprising the amino acid sequence of RASQGISSGSA (SEQ ID NO;

     12);

(e) a CDR-L2 comprising the amino acid sequence of DVSSLES (SEQ ID NO;

     7); and

(f) a CDR-L3 comprising the amino acid sequence of QQFNSYLLT (SEQ ID NO;

     8).

2. The antibody of claim 1, wherein the antibody comprises (a) a heavy chain variable domain (VH) sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:

13;

(b) a light chain variable domain (VL) sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO;

14;

or (c) a VH sequence as in (a) and a VL sequence as in (b).

4. The antibody of claim 1, wherein the antibody binds to an epitope comprising amino acid residues 71-105 of OspA (SEQ ID NO:

15).

9. The antibody of claim 1, wherein the antibody binds to three or more Borrelia species.

10. The antibody of claim 9, wherein the three or more Borrelia species comprises Borrelia burgdorferi, Borrelia afzelii, and Borrelia garinii.

11. The antibody of claim 10, wherein the antibody binds to OspA of Borrelia burgdorferi, Borrelia afzelii, and Borrelia garinii with a K_D of at least about 10 nM or lower.

12. The antibody of claim 9, wherein the antibody exhibits borreliacidal activity against Borrelia species comprising Borrelia burgdorferi, Borrelia afzelii, and Borrelia garinii in a borreliacidal assay.

29. The antibody of claim 1, wherein the antibody is monoclonal, human, humanized, or chimeric.

30. The antibody of claim 1, wherein the antibody is an antibody fragment that binds OspA.

31. The antibody of claim 30, wherein the antibody fragment is selected from the group consisting of Fab, Fab′

-SH, Fv, scFv, and (Fab′

)₂ fragments.

32. The antibody of claim 1, wherein the antibody is a full-length antibody.

33. The antibody of claim 1, wherein the half-life of the antibody is 3 days.

34. A pharmaceutical composition comprising the antibody of claim 1.

35. An isolated nucleic acid encoding the antibody of claim 1.

36. A vector comprising the nucleic acid of claim 35.

37. A host cell comprising the vector of claim 36.

38. A method of producing an antibody that specifically binds to outer surface protein A (OspA), the method comprising culturing the host cell of claim 37 in a culture medium.

39. A method of treating a subject having a disorder associated with a Borrelia infection comprising administering to the subject a therapeutically effective amount of the antibody of claim 1, thereby treating the subject.

40. A method of treating a subject at risk of developing a disorder associated with a Borrelia infection comprising administering to the subject a therapeutically effective amount of the antibody of claim 1, thereby treating the subject.

41. The method of claim 39, wherein the disorder associated with a Borrelia infection is Lyme borreliosis (Lyme disease).

42. A method of testing a Borrelia vector for the presence of Borrelia, the method comprising contacting a sample from the Borrelia vector with the antibody of claim 1 and determining if binding occurs, wherein binding is indicative of the presence of a Borrelia.

43. A method of decreasing Borrelia load in a Borrelia vector, the method comprising providing the antibody of claim 1 to the Borrelia vector.

44. A method of detecting a Borrelia infection in a subject comprising contacting a body fluid of the subject with the antibody of claim 1 and determining if binding occurs, wherein binding is indicative of the presence of a Borrelia infection.

3. An isolated antibody that specifically binds to OspA, wherein the antibody comprises (a) a VH sequence comprising the amino acid sequence of SEQ ID NO:

13 and (b) a VL sequence comprising the amino acid sequence of SEQ ID NO;

14.

5. An isolated antibody that specifically binds to OspA, wherein the antibody comprises the following six CDRs:

(a) a CDR-H1 comprising the amino acid sequence of GYKFSSYWIG (SEQ ID NO;

     16);

(b) a CDR-H2 comprising the amino acid sequence of IIFPGDSDKRYSPSFQG (SEQ ID NO;

     17);

(c) a CDR-H3 comprising the amino acid sequence of ARHITTHTYRGFFDF (SEQ ID NO;

     5);

(d) a CDR-L1 comprising the amino acid sequence of RASQDISSALA (SEQ ID NO;

     18);

(e) a CDR-L2 comprising the amino acid sequence of DVSSLES (SEQ ID NO;

     7); and

(f) a CDR-L3 comprising the amino acid sequence of QQFNGYPHRLT (SEQ ID NO;

     9).

6. The antibody of claim 5, wherein the antibody comprises (a) a VH sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:

19;

(b) a VL sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO;

20;

or (c) a VH sequence as in (a) and a VL sequence as in (b).

8. The antibody of claim 5, wherein the antibody binds to an epitope comprising amino acid residues 106-141 of OspA (SEQ ID NO:

21).

7. An isolated antibody that specifically binds to OspA, wherein the antibody comprises (a) a VH sequence comprising the amino acid sequence of SEQ ID NO:

19 and (b) a VL sequence comprising the amino acid sequence of SEQ ID NO;

20.

13. An isolated antibody that specifically binds to OspA, wherein the antibody comprises the following six CDRs:

(a) a CDR-H1 comprising the amino acid sequence of GYIFATYWIG (SEQ ID NO;

     23);

(b) a CDR-H2 comprising the amino acid sequence of IIYPNDSDTRYSPSFQG (SEQ ID NO;

     24);

(c) a CDR-H3 comprising the amino acid sequence of ARTRWYFDL (SEQ ID NO;

     25);

(d) a CDR-L1 comprising the amino acid sequence of RASQSVSSSYLA (SEQ ID NO;

     26);

(e) a CDR-L2 comprising the amino acid sequence of GASSRAT (SEQ ID NO;

     27); and

(f) a CDR-L3 comprising the amino acid sequence of QQYGSSPLT (SEQ ID NO;

     28).

14. The antibody of claim 13, wherein the antibody binds to an epitope comprising amino acid residues 178-273 of OspA (SEQ ID NO:

22).

15. The antibody of claim 13, wherein the antibody comprises (a) a heavy chain variable domain (VH) sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:

29;

(b) a light chain variable domain (VL) sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO;

30;

or (c) a VH sequence as in (a) and a VL sequence as in (b).

17. The antibody of claim 13, wherein the antibody binds to OspA of two or more Borrelia species.

18. The antibody of claim 17, wherein the two or more Borrelia species comprises Borrelia burgdorferi and Borrelia afzelii.

19. The antibody of claim 18, wherein the antibody binds to OspA of Borrelia burgdorferi and Borrelia afzelii with a K_D of at least about 1.2 μ

M or lower.

20. The antibody of claim 17, wherein the antibody exhibits borreliacidal activity against Borrelia species comprising Borrelia burgdorferi and Borrelia afzelii in a borreliacidal assay.

16. An isolated antibody that specifically binds to OspA, wherein the antibody comprises (a) a VH sequence comprising the amino acid sequence of SEQ ID NO:

29 and (b) a VL sequence comprising the amino acid sequence of SEQ ID NO;

30.

21. An isolated antibody that specifically binds to OspA, wherein the antibody comprises the following six CDRs:

(a) a CDR-H1 comprising the amino acid sequence of GFTFRNYWMD (SEQ ID NO;

     32);

(b) a CDR-H2 comprising the amino acid sequence of NIKQDGSVKYYVDSVEG (SEQ ID NO;

     33);

(c) a CDR-H3 comprising the amino acid sequence of ARDGYSGYDSVGFDI (SEQ ID NO;

     34);

(d) a CDR-L1 comprising the amino acid sequence of RASQSVSSSYLA (SEQ ID NO;

     35);

(e) a CDR-L2 comprising the amino acid sequence of DTSSRAT (SEQ ID NO;

     36); and

(f) a CDR-L3 comprising the amino acid sequence of QQYGSSPYT (SEQ ID NO;

     37).

22. The antibody of claim 21, wherein the antibody comprises (a) a heavy chain variable domain (VH) sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO:

38;

(b) a light chain variable domain (VL) sequence having at least 90% sequence identity to the amino acid sequence of SEQ ID NO;

39;

or (c) a VH sequence as in (a) and a VL sequence as in (b).

24. The antibody of claim 21, wherein the antibody binds to an epitope comprising amino acid residues 142-177 of OspA (SEQ ID NO:

31).

25. The antibody of claim 21, wherein the antibody binds to OspA of two or more Borrelia species.

26. The antibody of claim 25, wherein the one or more Borrelia species comprises Borrelia burgdorferi.

27. The antibody of claim 25, wherein the antibody binds to OspA of Borrelia burgdorferi with a K_D of at least about 5 nM or lower.

28. The antibody of claim 25, wherein the antibody exhibits borreliacidal activity against Borrelia species comprising Borrelia burgdorferi in a borreliacidal assay.

23. An isolated antibody that specifically binds to OspA, wherein the antibody comprises (a) a VH sequence comprising the amino acid sequence of SEQ ID NO:

38 and (b) a VL sequence comprising the amino acid sequence of SEQ ID NO;

39.