Method for virus propagation
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.
1. A method for rotavirus (RV) production comprising the following steps: cultivating cells in a bioreactor, infecting the cells with RV by activation with recombinant trypsin, and propagating the RV within said bioreactor in the presence of recombinant trypsin for maintenance/propagation, wherein the trypsin concentration is about 10 μg/ml for RV activation and about 5 μg/ml for maintenance/propagation.
2. The method according to claim 1 , wherein the cell cultivation is performed in VACCINEXPRESS medium.
3. The method according to claim 1 , wherein the bioreactor is a T-flask or spinner flask.
4. 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.
5. The method according to claim 1 , wherein the cell cultivation is performed on microcarriers.
6. The method according to claim 1 , wherein the microcarriers are sterilized microcarriers, preferably Cytodex gamma.
7. A method for rotavirus (RV) production comprising the following steps: cultivating cells in a bioreactor, infecting the cells with RV by activation with recombinant trypsin, and propagating the RV within said bioreactor in the presence of recombinant trypsin for maintenance/propagation, wherein the cell cultivation is performed on microcarriers, and wherein the trypsin concentration is about 20 μg/ml for RV activation and about 7 μg/ml for maintenance/propagation.
8. The method according to claim 1 , wherein the cells are Vero cells.
9. Use of the RV produced according to claim 1 for formulation of a vaccine against rotavirus caused disease.
10. Use of the RV produced according to claim 1 as a viral vector for gene therapy.