# US 8,465,742 B2 · Anti-cobra toxin antibody fragments and method of producing a VHH library

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- Number: US 8,465,742 B2
- Document ID: US8465742B2
- Status: Granted
- Kind: B2
- Application number: 13/132,161
- Filed: 2009-12-04
- Published: 2012-01-05
- Granted: 2013-06-18
- Earliest claimed: 2008-12-05
- Assignees: University of Guelph
- Applicants: J. Christopher Hall; Gabrielle Richard; Michael D. McLean
- Inventors: J. Christopher Hall; Gabrielle Richard; Michael D. McLean
- Web page: https://patentgrep.com/patent/US8465742B2

## CPC

- C07K 16/18: against material from animals or humans
- A61P 39/02: Antidotes
- A61K 2039/505: {comprising antibodies}
- C07K 2317/22: from camelids, e.g. camel, llama or dromedary
- C07K 2317/56: variable (Fv) region, i.e. VH and/or VL
- C07K 2317/565: Complementarity determining region [CDR]
- C07K 2317/569: Single domain, e.g. dAb, sdAb, VHH, VNAR or nanobody®
- 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
- Y10S 530/858: insects; venom
- 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)
- Y10S: TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS

## Abstract

A method of constructing a VHH library from an immunized camelid, using whole venom or an extract thereof There is also provided VHH antibody fragments isolated from a library produced in hyperimmunized llama These VHH antibody fragments were sequenced, and specifically bind α-cobratoxin.

## Claims

### Claim 1

An isolated VHH antibody fragment obtained from a VHH library prepared by a method comprising:
(1) immunizing a camelid with whole venom or an extract thereof;
(2) isolating nucleic acid sequences encoding the variable heavy fragment from the immunized camelid; and
(3) transforming a suitable host with the nucleic acid sequences to prepare a recombinant VHH library comprising VHH antibody fragments, wherein the VHH antibody fragments comprise a heavy chain complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO:5; a heavy chain complementarity determining region 2 (CDR2) comprising the amino acid sequence of SEQ ID NO:9; and a heavy chain complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO:11.

### Claim 2 (depends on 1)

The isolated VHH antibody fragment according to claim 1 that can bind to one or more proteins in a venom.

### Claim 3 (depends on 1)

The isolated VHH antibody fragment according to claim 1, wherein the venom is snake venom.

### Claim 4 (depends on 3)

The isolated VHH antibody fragment according to claim 3, wherein the VHH antibody fragment binds to α-cobratoxin.

### Claim 5

An isolated VHH antibody fragment comprising a heavy chain complementarity determining region 1 (CDR1) comprising the amino acid sequence of SEQ ID NO:5; a heavy chain complementarity determining region 2 (CDR2) comprising the amino acid sequence of SEQ ID NO:9; and a heavy chain complementarity determining region 3 (CDR3) comprising the amino acid sequence of SEQ ID NO:11.

### Claim 6

An isolated VHH antibody fragment comprising the amino acid sequence of SEQ ID NO:1.

### Claim 7 (depends on 5)

A pharmaceutical composition comprising an effective amount of the VHH antibody fragment according to claim 5 in admixture with a suitable diluent or carrier.

### Claim 8 (depends on 5)

A kit comprising an effective amount of the VHH antibody fragment according to claim 5 together with ancillary agents and instructions for use thereof.

### Claim 9 (depends on 8)

The kit according to claim 8, wherein the instructions for use thereof comprise treating a subject exposed to a venom and/or treating envenomation in a subject and/or neutralizing a venom in a subject that has been exposed to the venom.

### Claim 10 (depends on 6)

A pharmaceutical composition comprising an effective amount of the VHH antibody fragment according to claim 6 in admixture with a suitable diluent or carrier.

### Claim 11 (depends on 6)

A kit comprising an effective amount of the VHH antibody fragment according to claim 6 together with ancillary agents and instructions for use thereof.

### Claim 12 (depends on 11)

The kit according to claim 11, wherein the instructions for use thereof comprise treating a subject exposed to a venom and/or treating envenomation in a subject and/or neutralizing a venom in a subject that has been exposed to the venom.

### Claim 13 (depends on 5)

The isolated VHH antibody fragment according to claim 5 that can bind to one or more proteins in a venom.

### Claim 14 (depends on 5)

The isolated VHH antibody fragment according to claim 5, wherein the venom is snake venom.

### Claim 15 (depends on 5)

The isolated VHH antibody fragment according to claim 5, wherein the VHH antibody fragment binds to α-cobratoxin.