IP Library › Granted Patent US 7,700,738
Granted Patent B2
US 7,700,738 · App. 11/342,271 · Granted Apr 20, 2010

Therapeutic monoclonal antibodies that neutralize botulinum neurotoxins

Assignee: The Regents of the University of California
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Quick Facts
Patent No.
US 7,700,738
App. No.
11/342,271
Granted
Apr 20, 2010
Kind
B2
Abstract

This invention provides antibodies that specifically bind to and neutralize botulinum neurotoxin type A (BoNT/A) and the epitopes bound by those antibodies. The antibodies and derivatives thereof and/or other antibodies that specifically bind to the neutralizing epitopes provided herein can be used to neutralize botulinum neurotoxin and are therefore also useful in the treatment of botulism.

Claims (134)

1. A method of neutralizing Botulinum neurotoxin (BoNT) in a mammal said method comprising administering to said mammal at least two different antibodies for a BoNT serotype, wherein a first antibody of said two different antibodies comprises:

a) a V H CDR1 comprising the amino acid sequence of SEQ ID NO:49;

b) a V H CDR2 comprising the amino acid sequence of SEQ ID NO:51;

c) a V H CDR3 comprising the amino acid sequence of SEQ ID NO:75;

d) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105, or SEQ ID NO:109;

e) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:99, SEQ ID NO:107, or SEQ ID NO:111; and

f) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131 or SEQ ID NO:134.

2. The method of claim 1 , wherein said BoNT serotype is BoNT/A.

3. The method of claim 1 , wherein said first antibody binds at least two different subtypes selected from the group consisting of BoNT/A1, BoNT/A2, and BoNT/A3 each with an affinity greater than about 10 nM.

4. The method of claim 1 , wherein said first antibody binds BoNT/A1 and BoNT/A2 each with an affinity greater than about 10 nM.

5. The method of claim 4 , wherein the at least two different antibodies simultaneously bind at least one of subtypes BoNT/A1 and BoNT/A2.

6. The method of claim 1 , wherein said first antibody comprises:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:99; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131.

7. The method of claim 6 , wherein said first antibody comprises:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:13.

8. The method of claim 1 , wherein said first antibody comprises:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:107; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131.

9. The method of claim 8 , wherein said first antibody comprises:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:14.

10. The method of claim 1 , wherein said first antibody comprises:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:109;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:111; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:134.

11. The method of claim 10 , wherein said first antibody comprises:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:15.

12. The method of claim 1 , wherein at least one of said two different antibodies is a single chain Fv (scFv).

13. The method of claim 1 , wherein at least one of said two different antibodies is an IgG.

14. The method of claim 1 , wherein at least one of said two different antibodies is a Fab.

15. The method of claim 1 , wherein at least one of said two different antibodies is a (Fab′) 2 .

16. The method of claim 1 , wherein at least one of said two different antibodies is a (scFv′) 2 .

17. The method of claim 1 , wherein a second antibody of said two different antibodies is selected from the group consisting of:

a) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:11 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:10 or SEQ ID NO:354;

b) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 or SEQ ID NO:23 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:22;

c) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:12, and ii) a light chain comprising the amino acid sequence of SEQ ID NO:358, SEQ ID NO:13, SEQ ID NO:14, or SEQ ID NO:15; or

d) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:16 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:362.

18. The method of claim 1 , wherein a second antibody of said two different antibodies is selected from the group consisting of:

a) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:11 and ii) a light chain comprising the amino acid sequence of SEQ ID NO: 10 or SEQ ID NO:354;

b) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 or SEQ ID NO:23 and ii) a light chain comprising the amino acid sequence of SEQ ID N022 or

c) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:16 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:362.

19. A composition for neutralizing a Botulinum neurotoxin (BoNT), said composition comprising at least two different antibodies for a BoNT serotype, wherein a first antibody of said two different antibodies comprises:

a) a V H CDR1 comprising the amino acid sequence of SEQ ID NO:49;

b) a V H CDR2 comprising the amino acid sequence of SEQ ID NO:51;

c) a V H CDR3 comprising the amino acid sequence of SEQ ID NO:75;

d) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105 or SEQ ID

NO:109;

e) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:99, SEQ ID NO:107, or SEQ ID NO:111; and

f) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131 or SEQ ID

NO:134.

20. The composition of claim 19 , wherein said BoNT serotype is BoNT/A.

21. The composition of claim 19 , wherein said first antibody binds at least two different subtypes selected from the group consisting of BoNT/A1, BoNT/A2, and BoNT/A3 each with an affinity greater than about 10 nM.

22. The composition of claim 19 , wherein said first antibody binds BoNT/A1 and BoNT/A2 each with an affinity greater than about 10 nM.

23. The composition of claim 19 , wherein the at least two different antibodies simultaneously bind at least one of subtypes BoNT/A1 and BoNT/A2.

24. The composition of claim 19 , wherein said first antibody comprises:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:107; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131.

25. The composition of claim 24 , wherein said first antibody comprises:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:14.

26. The composition of claim 19 , wherein at least one of said two different antibodies is a single chain Fv (scFv).

27. The composition of claim 19 , wherein at least one of said two different antibodies is an IgG.

28. The composition of claim 19 , wherein at least one of said two different antibodies is a Fab.

29. The composition of claim 19 , wherein at least one of said two different antibodies is a (Fab′) 2 .

30. The composition of claim 19 , wherein at least one of said two different antibodies is a (scFv′) 2 .

31. The composition of claim 19 , wherein a second antibody of said two different antibodies is selected from the group consisting of:

a) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 11 and ii) a light chain comprising the amino acid sequence of SEQ ID NO: 10 or SEQ ID NO:354;

b) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 or SEQ ID NO:23 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:22;

c) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:12 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:358, SEQ ID NO:13, SEQ ID NO:14, or SEQ ID NO:15; or

d) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 16 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:362.

32. The composition of claim 19 , wherein a second antibody of said two different antibodies is selected from the group consisting of:

a) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:11 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:10 or SEQ ID NO:354;

b) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO:9 or SEQ ID NO:23 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:22; or

c) an antibody comprising i) a heavy chain comprising the amino acid sequence of SEQ ID NO: 16 and ii) a light chain comprising the amino acid sequence of SEQ ID NO:362.

33. The composition of claim 19 , wherein said antibodies are in a pharmaceutically acceptable excipient.

34. The composition of claim 33 , wherein said composition is a unit dosage formulation.

35. An isolated antibody that binds Botulinum neurotoxin (BoNT), wherein said antibody comprises:

a) a V H CDR1 comprising the amino acid sequence of SEQ ID NO:49;

b) a V H CDR2 comprising the amino acid sequence of SEQ ID NO:51;

c) a V H CDR3 comprising the amino acid sequence of SEQ ID NO:75;

d) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105 or SEQ ID NO:109;

e) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:99, SEQ ID NO:107, or SEQ ID NO:111; and

f) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131 or SEQ ID NO:134.

36. The antibody of claim 35 , wherein said antibody binds both the BoNT/A1 and BONT/A2 subtypes each with an affinity greater than about 10 nM.

37. The isolated antibody of claim 35 , wherein said antibody comprises:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:107; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131.

38. The isolated antibody of claim 37 , wherein said antibody comprises:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:14.

39. The antibody of claim 35 , wherein said antibody is a single chain Fv (scFv).

40. The antibody of claim 35 , wherein said antibody is an IgG.

41. The antibody of claim 35 , wherein said antibody is a Fab.

42. The antibody of claim 35 , wherein said antibody is a (Fab′) 2 .

43. The antibody of claim 35 , wherein said antibody is a (scFv′) 2 .

44. A kit for neutralizing a Botulinum neurotoxin, said kit comprising:

a composition according to any of claims 19, 20-25 and 26-34; and

instructional materials teaching the use of said composition to neutralize a Botulinum neurotoxin.

45. The kit of claim 44 , wherein said composition is stored in a disposable syringe.

46. The composition of claim 19 , wherein said first antibody comprises:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:99; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131.

47. The composition of claim 46 , wherein said first antibody comprises:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:13.

48. The composition of claim 19 , wherein said first antibody comprises:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:109;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:111; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:134.

49. The composition of claim 48 , wherein said first antibody comprises:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:15.

50. The isolated antibody of claim 35 comprising:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:105;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:99; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:131.

51. The isolated antibody of claim 50 comprising:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:13.

52. The isolated antibody of claim 35 comprising:

a) a V L CDR1 comprising the amino acid sequence of SEQ ID NO:109;

b) a V L CDR2 comprising the amino acid sequence of SEQ ID NO:111; and

c) a V L CDR3 comprising the amino acid sequence of SEQ ID NO:134.

53. The isolated antibody of claim 52 comprising:

a) a heavy chain comprising the amino acid sequence of SEQ ID NO:12; and

b) a light chain comprising the amino acid sequence of SEQ ID NO:15.

Assignments (2)
CORRECTIVE ASSIGNMENT TO CORRECT THE THIRD ASSIGNOR'S NAME, PREVIOUSLY RECORDED AT REEL 018483 FRAME 0202. Recorded Sep 27, 2007
From: MARKS, JAMES D.; AMERSDORFER, PETER; GEREN, ISIN N.; LOU, JIANLONG; RAZAI, ALI; GARCIA, MARIA CONSUELO
To: REGENTS OF THE UNIVERISTY OF CALIFORNIA, THE
Reel/Frame 019903/0461 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 3, 2006
From: MARKS, JAMES D.; AMERSDORFER, PETER; GERIN, ISIN; LOU, JIANLONG; RAZAI, ALI; GARCIA, MARIA CONSUELO
To: REGENTS OF THE UNIVERSITY OF CALIFORNIA, THE
Reel/Frame 018483/0202 →
Continuity (2)
Provisional Application 6064825600 · Jan 27, 2005
Related Publication 20080124328A1 · May 29, 2008