Methods for cultivating cells, propagating and purifying...

C - Chemistry – Metallurgy – 12 – N

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C12N 7/01 (2006.01) A61K 39/145 (2006.01) C12N 7/00 (2006.01) C12N 7/02 (2006.01)

Patent

CA 2738022

A novel serum-free cell culture medium and methods for cultivating MDCK cells are presented. In particular, non-tumorigenic MDCK cells. Methods for producing influenza viruses (e.g., particularly cold- adapted, and/or temperature sensitive, and/or attenuated influenza viruses) that eliminate the need for a cell culture medium exchange step are provided. The novel medium and methods are useful to grow influenza viruses, in cell culture to high titer. The present invention further provides purification methods for purifying influenza viruses with high overall recovery of live virus and result in levels of host cell DNA (HCD), host cell protein (HCP) and non-specific endonuclease (e.g., Benzonase), which are below the specifications required by regulatory agencies.

La présente invention concerne un nouveau milieu de culture cellulaire exempt de sérum et des procédés de culture de cellules MDCK, en particulier des cellules MDCK non tumogéniques. La présente invention concerne aussi des procédés de production de certains virus de la grippe, tels que les virus de la grippe particulièrement adaptés au froid, et/ou sensibles à la chaleur, et/ou atténués, qui évitent d'avoir à changer de milieu de culture cellulaire. Le milieu et les procédés de l'invention conviennent particulièrement à la culture des virus de la grippe, avec un titre élevé de culture cellulaire. L'invention concerne également des procédés de purification permettant de purifier les virus de la grippe avec un niveau globalement élevé de récupération du virus vivant, donnant des niveaux d'ADN de cellule hôte ou "HDC" (Host Cell DNA), de protéine de cellule hôte ou "HCP" (Host Cell Protein) et d'endonucléase non spécifique telle que la Benzonase inférieurs aux spécifications imposées par les organismes de régulation.

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