# US 10,117,923 B2 · Production of an immunogen using a plant virus

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- Number: US 10,117,923 B2
- Document ID: US10117923B2
- Status: Granted
- Kind: B2
- Application number: 15/607,189
- Filed: 2017-05-26
- Published: 2017-09-14
- Granted: 2018-11-06
- Earliest claimed: 2012-11-07
- Assignees: Vacplanta Limited
- Applicants: Vacplanta Limited
- Inventors: Petr Alexeevich Ivanov; Tatiana Vladimirovna Gasanova; Natalia Vitalievna Petukhova
- Web page: https://patentgrep.com/patent/US10117923B2

## CPC

- A61K 39/12: Viral antigens
- C07K 14/005: from viruses
- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- C12N 15/8258: {for the production of oral vaccines (antigens) or immunoglobulins}
- A61K 2039/5256: {expressing foreign proteins}
- A61K 2039/5258: {Virus-like particles}
- C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation
- C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2770/00022: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C12N 2770/00023: Virus like particles [VLP]
- 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)
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided herein is a recombinant rod-shaped viral particle comprising a fusion protein comprising a coat protein for a rod-shaped plant virus and a epitope from a human and/or human pathogen, where the epitope is disposed on the outer surface of the viral particle. In certain cases, the amino acid sequence of the coat protein is at least 40% identical to the amino acid sequence of the Tobacco Mosaic Virus (TMV) coat protein.

## Claims

### Claim 1

A recombinant rigid, rod-shaped chimeric viral particle capable of replication in a plant cell comprising:
a fusion protein comprising:
a coat protein that is at least 80% identical to the amino acid sequence of a wild type Tobacco Mosaic Virus (TMV) coat protein, and
the influenza A virus M2e epitope of SEQ ID NO: 1, except that the positions corresponding to cys17 and cys19 of SEQ ID NO:1 in the epitope are each independently substituted by a serine or alanine; and
wherein said epitope is inserted into said coat protein between Ser155 and Gly156 of the TMV coat protein.

### Claim 2 (depends on 1)

The viral particle of claim 1, wherein said particle has been produced by a plant cell.

### Claim 3 (depends on 2)

The viral particle of claim 2, wherein said particle has been produced by a Nicotiana benthamiana plant.

### Claim 4 (depends on 1)

A method of producing an immune response, comprising:
administering a recombinant rigid, rod-shaped viral particle of claim 1 to an animal, thereby causing the animal to produce antibodies that recognize said epitope.

### Claim 5 (depends on 4)

The method of claim 4, wherein said administering protects against future infection by said human and/or animal pathogen.

### Claim 6 (depends on 4)

The method of claim 4 wherein said viral particle is administered to said animal in conjunction with an adjuvant.

### Claim 7 (depends on 4)

The method of claim 4, wherein the administering is parenterally, enterally or orally.

### Claim 8 (depends on 1)

The viral particle of claim 1, wherein the amino acid sequence of said coat protein is identical to the TMV-U1 coat protein.

### Claim 9 (depends on 1)

The viral particle of claim 1, wherein the positions corresponding to cys17 and cys19 of SEQ ID NO:1 in the epitope are each substituted by a serine.

### Claim 10 (depends on 1)

The viral particle of claim 1, wherein the positions corresponding to cys17 and cys19 of SEQ ID NO:1 in the epitope are each substituted by an alanine.