# US 11,053,509 B2 · Plant-produced chimaeric Orbivirus VLPs

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

- Number: US 11,053,509 B2
- Document ID: US11053509B2
- Status: Granted
- Kind: B2
- Application number: 16/095,264
- Filed: 2017-04-19
- Published: 2019-05-09
- Granted: 2021-07-06
- Earliest claimed: 2016-04-19
- Assignees: Csir; University of Cape Town; Onderstepoort Biological Products Soc Ltd
- Applicants: Csir; University of Cape Town; Onderstepoort Biological Products Soc Ltd
- Inventors: Albertha René Van Zyl; Ann Elizabeth Meyers; Daria Anna Rutkowska; Edward Peter Rybicki; Hester Catharina Stark; Martha Magaretha O'Kennedy; Nobalanda Betty Mokoena
- Web page: https://patentgrep.com/patent/US11053509B2

## CPC

- C12N 15/8205: {Agrobacterium mediated transformation}
- A61K 35/76: Viruses; Subviral particles; Bacteriophages
- A61K 39/12: Viral antigens
- A61P 31/14: for RNA viruses
- C12N 15/8258: {for the production of oral vaccines (antigens) or immunoglobulins}
- A61K 2039/5258: {Virus-like particles}
- C12N 2720/12134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- 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

## Abstract

This invention relates to a second generation, plant-produced synthetic Orbivirus candidate vaccine. The vaccine comprises a plant produced chimaeric Orbivirus virus like particle (VLP) comprising at least one structural protein from one Orbivirus serotype and at least one structural protein selected from another serotype of the Orbivirus, wherein both structural capsid proteins are from the same Orbivirus species. In particular the invention relates to a vaccine against an Orbivirus, a method of producing chimaeric Orbivirus virus-like particles (VLPs) for use in a method of prevention and/or treatment of an Orbivirus infection, the use of the chimaeric Orbivirus VLPs in the manufacture of a vaccine for an Orbivirus, and a method of preventing and/or treating an Orbivirus infection.

## Claims

### Claim 1

A method of producing a chimaeric African Horse Sickness Virus (AHSV) VLP in a plant cell, the method comprising:
(i) providing codon-optimised nucleotide sequences encoding AHSV VP2, VP3, VP5 and VP7 structural proteins, wherein at least one of the VP2, VP3, VP5 and VP7 structural proteins is selected from a first AHSV serotype and at least one of the VP2, VP3, VP5 and VP7 structural proteins is selected from a second AHSV serotype of the same AHSV species;
(ii) cloning the codon-optimised nucleotide sequences into at least one expression vector adapted to express the structural proteins in a plant cell;
(iii) transforming or infiltrating the plant cell with the at least one expression vector of step (ii);
(iv) co-expressing the VP2, VP3, VP5 and VP7 structural proteins in the plant cell, such that the expressed structural proteins assemble to form the chimaeric AHSV VLP; and
(v) recovering the chimaeric AHSV VLP from the plant cell.

### Claim 2 (depends on 1)

The method of claim 1, wherein in step (i) at least one of the VP2, VP3, VP5 and VP7 structural proteins is selected from a third AHSV serotype.

### Claim 3 (depends on 2)

The method of claim 2, wherein at least one of the VP2, VP3, VP5 and VP7 structural proteins is selected from a fourth AHSV serotype.

### Claim 4 (depends on 1)

The method of claim 1, wherein the structural proteins are transiently expressed in the plant cell.

### Claim 5 (depends on 1)

The method of claim 1, wherein the at least one expression vector includes a promoter and/or other regulatory sequences, operably linked to each nucleotide sequence encoding each structural protein.

### Claim 6 (depends on 1)

The method of claim 1, wherein in step (iii) the at least one expression vector is transformed into the plant cell in a ratio of 1:1:1:1 or a ratio of 1:1:2:1 or a ratio of 2:1:2:1 of the nucleotide sequences encoding VP2:VP3:VP5:VP7.

### Claim 7 (depends on 1)

The method of claim 1, wherein the plant cell is a Nicotiana benthamiana cell.

### Claim 8 (depends on 1)

The method of claim 1, wherein the plant cell is a mutant N. benthamiana dXT/FT tobacco cell, which facilitates mammalian-like or human-like glycosylation of polypeptides.

### Claim 9 (depends on 1)

The method of claim 1, wherein the expression of the AHSV VP2, VP3, VP5 and VP7 structural proteins in the plant cell is mediated by Agrobacterium AGL-1, LBA4404 or GV3101 pMP90.