Antibodies that neutralize botulinum neurotoxins

Patent metadata

Field	Value
Publication number	US-9453068-B2
Application number	US-201514634355-A
Country	US
Kind code	B2
Filing date	Feb 27, 2015
Priority date	Jul 31, 2008
Publication date	Sep 27, 2016
Grant date	Sep 27, 2016

Abstract

This disclosure provides antibodies that specifically bind to and typically neutralize botulinum neurotoxins (e.g., BoNT/A, BoNT/B, BoNT/E, etc.) 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.

First claim

What is claimed is:

1. An isolated antibody that competes for binding to a Botulinum neurotoxin with an antibody comprising:
a V H CDR1 comprising the amino acid sequence NYPMS (SEQ ID NO:520);
a V H CDR2 comprising the amino acid sequence SLTASGDNTFYADSVKG (SEQ ID NO:522);
a V H CDR3 comprising the amino acid sequence of ALVGRYDISTGYYRPVMDS (SEQ ID NO:524);
a V L CDR1 comprising the amino acid sequence of RTSQGFTSALA (SEQ ID NO:770);
a V L CDR2 comprising the amino acid sequence of DASKLES (SEQ ID NO:772); and
a V L CDR3 comprising the amino acid sequence of QQSNSYPLT (SEQ ID NO:774).

2. The isolated antibody of claim 1 , wherein the antibody specifically binds an epitope of a Botulinum neurotoxin that is specifically bound by an antibody comprising:
a V H CDR1 comprising the amino acid sequence NYPMS (SEQ ID NO:520);
a V H CDR2 comprising the amino acid sequence SLTASGDNTFYADSVKG (SEQ ID NO:522);
a V H CDR3 comprising the amino acid sequence ALVGRYDISTGYYRPVMDS (SEQ ID NO:524);
a V L CDR1 comprising the amino acid sequence RTSQGFTSALA (SEQ ID NO:770);
a V L CDR2 comprising the amino acid sequence DASKLES (SEQ ID NO:772); and
a V L CDR3 comprising the amino acid sequence QQSNSYPLT (SEQ ID NO:774).

3. The isolated antibody of claim 1 , wherein the antibody comprises:
a variable heavy chain (V H ) comprising the amino acid sequence of SEQ ID NO:23; and
a variable light chain (V L ) comprising:
a V L CDR1 comprising the amino acid sequence RTSQGFTSALA (SEQ ID NO:770);
a V L CDR2 comprising the amino acid sequence DASKLES (SEQ ID NO:772); and
a V L CDR3 comprising the amino acid sequence QQSNSYPLT (SEQ ID NO:774).

4. The isolated antibody of claim 3 , wherein the V L comprises the amino acid sequence of SEQ ID NO:59.

5. The isolated antibody of claim 1 , wherein the antibody comprises:
a variable heavy chain (V H ) comprising:
a V H CDR1 comprising the amino acid sequence NYPMS (SEQ ID NO:520);
a V H CDR2 comprising the amino acid sequence SLTASGDNTFYADSVKG (SEQ ID NO:522); and
a V H CDR3 comprising the amino acid sequence ALVGRYDISTGYYRPVMDS (SEQ ID NO:524); and

a variable light chain (V L ) comprising the amino acid sequence of SEQ ID NO:59.

6. The isolated antibody of claim 5 , wherein the V H comprises the amino acid sequence of SEQ ID NO:23.

7. The isolated antibody of claim 2 , wherein the antibody comprises:
a variable heavy chain (V H ) comprising the amino acid sequence of SEQ ID NO:23; and
a variable light chain (V L ) comprising:
a V L CDR1 comprising the amino acid sequence RTSQGFTSALA (SEQ ID NO:770);
a V L CDR2 comprising the amino acid sequence DASKLES (SEQ ID NO:772); and
a V L CDR3 comprising the amino acid sequence QQSNSYPLT (SEQ ID NO:774).

8. The isolated antibody of claim 7 , wherein the V L comprises the amino acid sequence of SEQ ID NO:59.

9. The isolated antibody of claim 2 , wherein the antibody comprises:
a variable heavy chain (V H ) comprising:
a V H CDR1 comprising the amino acid sequence NYPMS (SEQ ID NO:520);
a V H CDR2 comprising the amino acid sequence SLTASGDNTFYADSVKG (SEQ ID NO:522); and
a V H CDR3 comprising the amino acid sequence ALVGRYDISTGYYRPVMDS (SEQ ID NO:524); and

a variable light chain (V L ) comprising the amino acid sequence of SEQ ID NO:59.

10. The isolated antibody of claim 9 , wherein the V H comprises the amino acid sequence of SEQ ID NO:23.

11. The isolated antibody of claim 1 , wherein the antibody is humanized.

12. The isolated antibody of claim 1 , wherein the antibody is selected from the group consisting of: a single chain Fv (scFv), an IgG, a Fab, a (Fab′) 2 , and an (scFv′) 2 .

13. The isolated antibody of claim 1 , wherein the antibody is an IgG.

14. The isolated antibody of claim 2 , wherein the antibody is humanized.

15. The isolated antibody of claim 2 , wherein the antibody is selected from the group consisting of: a single chain Fv (scFv), an IgG, a Fab, a (Fab′) 2 , and an (scFv′) 2 .

16. The isolated antibody of claim 2 , wherein the antibody is an IgG.

17. A conjugate comprising:
the antibody of claim 1 ; and
a label.

18. The conjugate of claim 17 , wherein the label is selected from the group consisting of: a fluorescent label, a spectroscopic label, an enzyme, a colorimetric label, a chemiluminescent label, biotin, and streptavidin.

19. The conjugate of claim 17 , wherein the antibody specifically binds an epitope of a Botulinum neurotoxin that is specifically bound by an antibody comprising:
a V H CDR1 comprising the amino acid sequence NYPMS (SEQ ID NO:520);
a V H CDR2 comprising the amino acid sequence SLTASGDNTFYADSVKG (SEQ ID NO:522);
a V H CDR3 comprising the amino acid sequence of ALVGRYDISTGYYRPVMDS (SEQ ID NO:524);
a V L CDR1 comprising the amino acid sequence of RTSQGFTSALA (SEQ ID NO:770);
a V L CDR2 comprising the amino acid sequence of DASKLES (SEQ ID NO:772); and
a V L CDR3 comprising the amino acid sequence of QQSNSYPLT (SEQ ID NO:774).

20. A kit comprising the conjugate of claim 17 .

21. A pharmaceutical composition comprising:
the antibody of claim 1 ; and
a pharmaceutically acceptable carrier.

22. The pharmaceutical composition of claim 21 , wherein the antibody specifically binds an epitope of a Botulinum neurotoxin that is specifically bound by an antibody comprising:
a V H CDR1 comprising the amino acid sequence of SEQ ID NO:520;
a V H CDR2 comprising the amino acid sequence of SEQ ID NO:522;
a V H CDR3 comprising the amino acid sequence of SEQ ID NO:524;
a V L CDR1 comprising the amino acid sequence of SEQ ID NO:770;
a V L CDR2 comprising the amino acid sequence of SEQ ID NO:772; and
a V L CDR3 comprising the amino acid sequence of SEQ ID NO:774.

23. A kit comprising the pharmaceutical composition of claim 21 .

24. A method of specifically binding a Botulinum neurotoxin in a mammal comprising:
administering to the mammal an effective amount of the antibody of claim 1 ,
wherein the administering provides for binding of the antibody to the botulinum neurotoxin present in the mammal.

25. The method according to claim 24 , wherein the antibody specifically binds an epitope of a Botulinum neurotoxin that is specifically bound by an antibody comprising:
a V H CDR1 comprising the amino acid sequence NYPMS (SEQ ID NO:520);
a V H CDR2 comprising the amino acid sequence SLTASGDNTFYADSVKG (SEQ ID NO:522);
a V H CDR3 comprising the amino acid sequence of ALVGRYDISTGYYRPVMDS (SEQ ID NO:524);
a V L CDR1 comprising the amino acid sequence of RTSQGFTSALA (SEQ ID NO:770);
a V L CDR2 comprising the amino acid sequence of DASKLES (SEQ ID NO:772); and
a V L CDR3 comprising the amino acid sequence of QQSNSYPLT (SEQ ID NO:774).

26. A method of determining the presence or absence of specifically binding a Botulinum neurotoxin in a biological sample, comprising:
contacting the biological sample with the conjugate of claim 17 .

27. The method according to claim 26 , wherein the antibody specifically binds an epitope of a Botulinum neurotoxin that is specifically bound by an antibody comprising:
a V H CDR1 comprising the amino acid sequence NYPMS (SEQ ID NO:520);
a V H CDR2 comprising the amino acid sequence SLTASGDNTFYADSVKG (SEQ ID NO:522);
a V H CDR3 comprising the amino acid sequence of ALVGRYDISTGYYRPVMDS (SEQ ID NO:524);
a V L CDR1 comprising the

Assignees

• Univ California

Inventors

• Marks James D
• Lou Jianlong

Classifications

• A61P31/04

  Antibacterial agents · CPC title

• A61P39/02

  Antidotes · CPC title

• C07K2317/52

  Constant or Fc region; Isotype · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• C07K16/1282 Primary

  Clostridium (G) · CPC title