# US 10,369,206 B2 · Clostridium difficile antigens

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- Number: US 10,369,206 B2
- Document ID: US10369206B2
- Status: Granted
- Kind: B2
- Application number: 13/878,150
- Filed: 2011-10-05
- Published: 2013-10-10
- Granted: 2019-08-06
- Earliest claimed: 2010-10-05
- Assignees: The Secretary of State for Health; MicroPharm Limited
- Applicants: Clifford Shone; April Roberts; Helen Ahern; Michael Maynard-Smith; John Landon
- Inventors: Clifford Shone; April Roberts; Helen Ahern; Michael Maynard-Smith; John Landon
- Web page: https://patentgrep.com/patent/US10369206B2

## CPC

- A61K 39/08: Clostridium, e.g. Clostridium tetani
- A61K 38/164: {from bacteria}
- A61K 39/40: bacterial
- A61P 31/04: Antibacterial agents
- C07K 14/33: from Clostridium (G)
- C07K 16/1282: Clostridium (G)
- A61K 2039/505: {comprising antibodies}
- C07K 2317/76: Antagonist effect on antigen, e.g. neutralization or inhibition of binding
- C07K 2319/00: Fusion polypeptide
- 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)
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- 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)

## Abstract

The present invention relates to recombinant Clostridium difficile antigens based on a fusion protein that consists of or comprises a first amino acid sequence and a second amino acid sequence, wherein: a) the first amino acid sequence is provided by an amino acid sequence that has at least 80% sequence identity with an amino acid sequence consisting of residues 500-1850 of a C. difficile Toxin A sequence or residues 1500-1851 of a C. difficile Toxin B sequence; and b) the second amino acid sequence is provided by an amino acid sequence that has at least 80% sequence identity with an amino acid sequence consisting of a long repeat unit located within amino acid residues 1851-2710 of a C. difficile Toxin A sequence or within amino acid residues 1852-2366 of a C. difficile Toxin B sequence; though with the proviso that the fusion protein is not a polypeptide comprising amino acid residues 543-2710 of a C. difficile Toxin A and with the proviso that the fusion protein is not a polypeptide comprising amino acid residues 543-2366 of a C. difficile Toxin B. Also provided is the use of said antigens for the prevention/treatment/suppression of Clostridium difficile infection (CDI), together with methods for generating said antigens, methods for generating antibodies that bind to said antigens, and the use of said antibodies for the prevention/treatment/suppression of CDI.

## Claims

### Claim 1

A protein, comprising
a) an amino acid sequence that comprises amino acid residues 767-2366 of a C. difficile Toxin B sequence; or
b) an amino acid sequence that comprises amino acid residues 1145-2366 of a C. difficile Toxin B sequence;
with the proviso that the protein is not a polypeptide comprising amino acid residues 543-2366 of the C. difficile Toxin B;
wherein the protein can elicit a toxin-neutralizing immune response; and
wherein said C. difficile Toxin B sequences are the C. difficile Toxin B amino acid residue sequence 767-2366 or 1145-2366 of SEQ ID NO:2 or the C. difficile Toxin B amino acid residue sequence 767-2366 or 1145-2366 of SEQ ID NO:4.

### Claim 2 (depends on 1)

The protein according to claim 1, wherein other than the above-defined Toxin B amino acid sequences, said protein does not include any additional Toxin B amino acid sequence.

### Claim 3 (depends on 1)

A method of generating an antibody that binds to a C. difficile Toxin B, the method comprising:
immunizing an animal with the protein according to claim 1 to induce production of an antibody specific for C. difficile Toxin B.

### Claim 4 (depends on 3)

The method according to claim 3, further comprising the step of isolating the antibody specific for C. difficile Toxin B from the immunized animal.

### Claim 5 (depends on 4)

The method according to claim 4, wherein the antibody is isolated by affinity purification.

### Claim 6 (depends on 3)

The method according to claim 3, wherein the animal is horse, sheep, goat, or human.

### Claim 7

A protein, consisting of:
a) amino acid residues 767-2366 of a C. difficile Toxin B sequence; or
b) amino acid residues 1145-2366 of a C. difficile Toxin B sequence;
wherein the protein can elicit a toxin-neutralizing immune response; and
wherein said C. difficile Toxin B sequences are the C. difficile Toxin B amino acid residue sequence 767-2366 or 1145-2366 of SEQ ID NO:2 or the C. difficile Toxin B amino acid residue sequence 767-2366 or 1145-2366 of SEQ ID NO:4.