# US 2020/0216820 A1 · Method for virus propagation

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- Number: US 2020/0216820 A1
- Document ID: US20200216820A1
- Status: Published application
- Kind: A1
- Application number: 16/737,390
- Filed: 2020-01-08
- Published: 2020-07-09
- Earliest claimed: 2019-01-08
- Assignees: GE Healthcare Bio-Sciences AB
- Applicants: GE Healthcare Bio-Sciences AB
- Inventors: Ann-Christin MAGNUSSON; Mats LUNDGREN; Eva BLANCK; Christine SUND LUNDSTROM
- Web page: https://patentgrep.com/patent/US20200216820A1

## CPC

- C12N 7/00: Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof (preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00)
- A61K 39/12: Viral antigens
- A61P 31/14: for RNA viruses
- C12N 5/0686: {Kidney cells}
- C12N 9/88: Lyases (4.)
- C12N 15/86: Viral vectors
- A61K 39/00: Medicinal preparations containing antigens or antibodies (materials for immunoassay G01N33/53)
- C12N 2720/12331: Uses of virus other than therapeutic or vaccine, e.g. disinfectant
- C12N 2720/12334: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
- C12N 2720/12343: viral genome or elements thereof as genetic vector
- C12N 2720/12351: Methods of production or purification of viral material
- 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

The present invention relates to a method for virus propagation. More closely the invention relates to a method for animal component free propagation and production of rotavirus (RV) using recombinant trypsin.

## Claims

### Claim 1

A method for rotavirus (RV) production comprising the following steps: cell cultivation in a bioreactor, infection of the cells with RV by activation with recombinant trypsin, and RV propagation within said bioreactor in the presence of recombinant trypsin for maintainance/propagation.

### Claim 2 (depends on 1)

The method according to claim 1, wherein the cell cultivation is performed in VaccinExpress medium.

### Claim 3 (depends on 1)

The method according to claim 1, wherein the bioreactor is a T-flask or spinner flask.

### Claim 4 (depends on 1)

The method according to claim 1, wherein the bioreactor is a disposable bag on a rocking platform, preferably a Wave bioreactor, or a stirred tank.

### Claim 5 (depends on 1)

The method according to claim 1, wherein the cell cultivation is performed on microcarriers.

### Claim 6 (depends on 1)

The method according to claim 1, wherein the microcarriers are sterilized microcarriers, preferably Cytodex gamma.

### Claim 7 (depends on 1)

The method according to claim 1, wherein the trypsin concentration is about 10 μg/ml for RV activation and about 5 μg/ml for maintenance/propagation.

### Claim 8 (depends on 5)

The method according to claim 5, wherein the trypsin concentration is about 20 μg/ml for RV activation and about 7 μg/ml for maintenance/propagation when microcarriers are used.

### Claim 9 (depends on 1)

The method according to claim 1, wherein the cells are Vero cells.

### Claim 10 (depends on 1)

Use of the RV produced according to claim 1 for formulation of a vaccine against rotavirus caused disease.

### Claim 11 (depends on 1)

Use of the RV produced according to claim 1 as a viral vector for gene therapy.

### Claim 12

Use of recombinant trypsin for virus infection of cells.