# US 10,308,708 B2 · Antibodies that neutralize botulinum neurotoxins

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 10,308,708 B2
- Document ID: US10308708B2
- Status: Granted
- Kind: B2
- Application number: 15/238,419
- Filed: 2016-08-16
- Published: 2017-04-06
- Granted: 2019-06-04
- Earliest claimed: 2008-07-31
- Assignees: The Regents of the University of California
- Applicants: The Regents of the University of California
- Inventors: James D. Marks; Isin N. Geren
- Web page: https://patentgrep.com/patent/US10308708B2

## CPC

- C07K 16/1282: Clostridium (G)
- A61P 31/04: Antibacterial agents
- A61P 39/02: Antidotes
- A61K 2039/505: {comprising antibodies}
- A61K 2039/507: {Comprising a combination of two or more separate antibodies}
- C07K 2317/21: from primates, e.g. man
- C07K 2317/52: Constant or Fc region; Isotype
- C07K 2317/54: F(ab')2
- C07K 2317/55: Fab or Fab'
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/622: Single chain antibody (scFv)
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2317/92: Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
- C07K 2319/00: Fusion polypeptide
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## 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.

## Claims

### Claim 1

A composition comprising:
a first nucleic acid comprising the isolated nucleic acid comprising a nucleotide sequence encoding a variable heavy chain (VH ) polypeptide comprising
a VH CDR1 comprising the amino acid sequence of HYGMH (SEQ ID NO:492);
a VH CDR2 comprising the amino acid sequence of VIWYDGRNPYYAASVKG (SEQ ID NO:494); and
a VH CDR3 comprising the amino acid sequence of DLTRFHDTTFGVFEM (SEQ ID NO:496); and
a second nucleic acid comprising the isolated nucleic acid comprising a nucleotide sequence encoding a variable light chain (VL ) polypeptide comprising
a VL CDR1 comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:742);
a VL CDR2 comprising the amino acid sequence of AASSLQS (SEQ ID NO:744); and
a VL CDR3 comprising the amino acid sequence of QQYSSLYT (SEQ ID NO:746);
wherein the first and second isolated nucleic acids are on the same or different nucleic acid constructs.

### Claim 2

A composition comprising:
a first nucleic acid comprising the isolated nucleic acid comprising a nucleotide sequence encoding a variable heavy chain (VH ) polypeptide comprising
a VH CDR1 comprising the amino acid sequence of GFTFSHYG (SEQ ID NO:1309);
a VH CDR2 comprising the amino acid sequence of IWYDGRNP (SEQ ID NO:1310);
and
a VH CDR3 comprising the amino acid sequence of VKDLTRFHDTTFGVFEM (SEQ ID NO:1311); and
a second nucleic acid comprising the isolated nucleic acid comprising a nucleotide sequence encoding a variable light chain (VL ) polypeptide comprising
a VL CDR1 comprising the amino acid sequence of QGISSW (SEQ ID NO:1312);
a VL CDR2 comprising the amino acid sequence of AAS (SEQ ID NO:1313); and
a VL CDR3 comprising the amino acid sequence of QQYSSL (SEQ ID NO:1314);
wherein the first and second isolated nucleic acids are on the same or different nucleic acid constructs.

### Claim 3

A composition comprising:
a first nucleic acid comprising the isolated nucleic acid comprising a nucleotide sequence encoding a variable heavy chain (VH ) polypeptide comprising
a VH CDR1 comprising the amino acid sequence of HYGMH (SEQ ID NO:492);
a VH CDR2 comprising the amino acid sequence of VIWYDGRNPYYAASVKG (SEQ
ID NO:494); and
a VH CDR3 comprising the amino acid sequence of DLTRFHDTTFGVFEM (SEQ ID NO:496); and
a second nucleic acid comprising the isolated nucleic acid comprising a nucleotide sequence encoding a variable light chain (VL ) polypeptide comprising
a VL CDR1 comprising the amino acid sequence of RASQSISSWLA (SEQ ID NO:735);
a VL CDR2 comprising the amino acid sequence of KASSLEN (SEQ ID NO:737); and
a VL CDR3 comprising the amino acid sequence of QQYSTYSRT (SEQ ID NO:739);
wherein the first and second isolated nucleic acids are on the same or different nucleic acid constructs.

### Claim 4

A composition comprising:
a first isolated nucleic acid comprising a nucleotide sequence encoding a variable heavy chain (VH ) polypeptide comprising the amino acid sequence of SEQ ID NO:19; and
a second isolated nucleic acid comprising a nucleotide sequence encoding a variable light chain (VL ) polypeptide comprising:
a VL CDR1 comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:742);
a VL CDR2 comprising the amino acid sequence of AASSLQS (SEQ ID NO:744); and
a VL CDR3 comprising the amino acid sequence of QQYSSLYT (SEQ ID NO:746); or
a VL CDR1 comprising the amino acid sequence of QGISSW (SEQ ID NO:1312);
a VL CDR2 comprising the amino acid sequence of AAS (SEQ ID NO:1313); and
a VL CDR3 comprising the amino acid sequence of QQYSSL (SEQ ID NO:1314); or
a VL CDR1 comprising the amino acid sequence of RASQSISSWLA (SEQ ID NO:735);
a VL CDR2 comprising the amino acid sequence of KASSLEN (SEQ ID NO:737); and
a VL CDR3 comprising the amino acid sequence of QQYSTYSRT (SEQ ID NO:739); or
the amino acid sequence of SEQ ID NO: 55; or
the amino acid sequence of SEQ ID NO: 54
wherein the first and second isolated nucleic acids are on the same or different nucleic acid constructs.

### Claim 5 (depends on 4)

The composition of claim 4, wherein the second isolated nucleic acid comprises a nucleotide sequence encoding a VL polypeptide comprising:
a VL CDR1 comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:742);
a VL CDR2 comprising the amino acid sequence of AASSLQS (SEQ ID NO:744); and
a VL CDR3 comprising the amino acid sequence of QQYSSLYT (SEQ ID NO:746).

### Claim 6 (depends on 4)

The composition of claim 4, wherein the second isolated nucleic acid comprises a nucleotide sequence encoding a VL polypeptide comprising:
a VL CDR1 comprising the amino acid sequence of QGISSW (SEQ ID NO:1312);
a VL CDR2 comprising the amino acid sequence of AAS (SEQ ID NO:1313); and
a VL CDR3 comprising the amino acid sequence of QQYSSL (SEQ ID NO:1314).

### Claim 7 (depends on 4)

The composition of claim 4, wherein the second isolated nucleic acid comprises a nucleotide sequence encoding a VL polypeptide comprising the amino acid sequence of SEQ ID NO: 55.

### Claim 8 (depends on 4)

The composition of claim 4, wherein the second isolated nucleic acid comprises a nucleotide sequence encoding a VL polypeptide comprising:
a VL CDR1 comprising the amino acid sequence of RASQSISSWLA (SEQ ID NO:735);
a VL CDR2 comprising the amino acid sequence of KASSLEN (SEQ ID NO:737); and
a VL CDR3 comprising the amino acid sequence of QQYSTYSRT (SEQ ID NO:739).

### Claim 9 (depends on 4)

The composition of claim 4, wherein the second isolated nucleic acid comprises a nucleotide sequence encoding a VL polypeptide comprising the amino acid sequence of SEQ ID NO: 54.

### Claim 10

A composition comprising:
a first isolated nucleic acid comprising a nucleotide sequence encoding a VL polypeptide comprising the amino acid sequence of SEQ ID NO:55; and
a second isolated nucleic acid comprising a nucleotide sequence encoding a VH polypeptide comprising:
a VH CDR1 comprising the amino acid sequence of HYGMH (SEQ ID NO:492);
a VH CDR2 comprising the amino acid sequence of VIWYDGRNPYYAASVKG (SEQ ID NO:494);
a VH CDR3 comprising the amino acid sequence of DLTRFHDTTFGVFEM (SEQ ID NO:496); or
a second isolated nucleic acid present on a nucleic acid construct, the second isolated nucleic acid comprising a nucleotide sequence encoding a VH polypeptide comprising:
a VH CDR1 comprising the amino acid sequence of GFTFSHYG (SEQ ID NO:1309);
a VH CDR2 comprising the amino acid sequence of IWYDGRNP (SEQ ID NO:1310);
a VH CDR3 comprising the amino acid sequence of VKDLTRFHDTTFGVFEM (SEQ ID NO:1311);
wherein the first and second isolated nucleic acids are on the same or different constructs.

### Claim 11 (depends on 10)

The composition of claim 10, wherein the second isolated nucleic acid comprises a nucleotide sequence encoding an amino acid sequence of a VH polypeptide comprising:
a VH CDR1 comprising the amino acid sequence of HYGMH (SEQ ID NO:492);
a VH CDR2 comprising the amino acid sequence of VIWYDGRNPYYAASVKG (SEQ ID NO:494); and
a VH CDR3 comprising the amino acid sequence of DLTRFHDTTFGVFEM (SEQ ID NO:496).

### Claim 12 (depends on 10)

The composition of claim 10, wherein the second isolated nucleic acid comprises a nucleotide sequence encoding an amino acid sequence of a VH polypeptide comprising:
a VH CDR1 comprising the amino acid sequence of GFTFSHYG (SEQ ID NO:1309);
a VH CDR2 comprising the amino acid sequence of IWYDGRNP (SEQ ID NO:1310); and
a VH CDR3 comprising the amino acid sequence of VKDLTRFHDTTFGVFEM (SEQ ID NO:1311).

### Claim 13

A nucleic acid construct comprising:
i) a first nucleic acid comprising a nucleotide sequence encoding an amino acid sequence of a variable heavy chain (VH ) polypeptide comprising
a VH CDR1 comprising the amino acid sequence of HYGMH (SEQ ID NO:492);
a VH CDR2 comprising the amino acid sequence of VIWYDGRNPYYAASVKG (SEQ ID NO:494); and
a VH CDR3 comprising the amino acid sequence of DLTRFHDTTFGVFEM (SEQ ID NO:496); or
a VH CDR1 comprising the amino acid sequence of GFTFSHYG (SEQ ID NO:1309);
a VH CDR2 comprising the amino acid sequence of IWYDGRNP (SEQ ID NO:1310); and
a VH CDR3 comprising the amino acid sequence of VKDLTRFHDTTFGVFEM (SEQ ID NO:1311); and
ii) a second nucleic acid comprising a nucleotide sequence encoding a variable light chain (VL ) polypeptide comprising:
a VL CDR1 comprising the amino acid sequence of RASQGISSWLA (SEQ ID NO:742);
a VL CDR2 comprising the amino acid sequence of AASSLQS (SEQ ID NO:744); and
a VL CDR3 comprising the amino acid sequence of QQYSSLYT (SEQ ID NO:746); or
a VL CDR1 comprising the amino acid sequence of QGISSW (SEQ ID NO:1312);
a VL CDR2 comprising the amino acid sequence of AAS (SEQ ID NO:1313); and
a VL CDR3 comprising the amino acid sequence of QQYSSL (SEQ ID NO:1314) or
a VL CDR1 comprising the amino acid sequence of RASQSISSWLA (SEQ ID NO:735);
a VL CDR2 comprising the amino acid sequence of KASSLEN (SEQ ID NO:737); and
a VL CDR3 comprising the amino acid sequence of QQYSTYSRT (SEQ ID NO:739).

### Claim 14 (depends on 1)

A cell comprising the first and second isolated nucleic acids of claim 1.

### Claim 15 (depends on 2)

A cell comprising the first and second isolated nucleic acids of claim 2.

### Claim 16 (depends on 3)

A cell comprising the first and second isolated nucleic acids of claim 3.

### Claim 17 (depends on 14)

A method of making a polypeptide, the method comprising culturing the cell of claim 14 under conditions suitable for the cell to express the polypeptide, wherein the polypeptide is produced.

### Claim 18 (depends on 15)

A method of making a polypeptide, the method comprising culturing the cell of claim 15 under conditions suitable for the cell to express the polypeptide, wherein the polypeptide is produced.

### Claim 19 (depends on 16)

A method of making a polypeptide, the method comprising culturing the cell of claim 16 under conditions suitable for the cell to express the polypeptide, wherein the polypeptide is produced.