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AssigneeCrucell Holland B.V.
InventorsShuyuan Zhang, Capucine Thwin, Zheng Wu, Toohyon Cho, Shawn Gallagher
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The present invention addresses the need to improve the yields of viral vectors when grown in cell culture systems. In particular, it has been demonstrated that for adenovirus, the use of low-medium perfusion rates in an attached cell culture system provides for improved yields. In other embodiments, the inventors have shown that there is improved Ad-p53 production with cells grown in serum-free conditions, and in particular in serum-free suspension culture. Also important to the increase of yields is the use of detergent lysis. Combination of these aspects of the invention permits purification of virus by a single chromatography step that results in purified virus of the same quality as preparations from double CsCl banding using an ultracentrifuge.
A process for preparing adenovirus, the process comprising:
A process for preparing adenovirus, the process comprising:
A method for producing adenovirus that includes culturing producer cells and infecting the cultured producer cells with an adenovirus, wherein the improvement comprises infecting said producer cells with the adenovirus when the cells in culture are between late-log phase of growth and stationary phase of growth.
In a method for producing adenovirus that includes culturing producer cells and infecting the cultured producer cells with an adenovirus, wherein the improvement comprises infecting said producer cells with the adenovirus when the cells in culture are between mid-log phase of growth and stationary phase of growth and the producer cells are essentially homogeneous with respect to the phase of cell growth.
Dependencies are inferred from the claim text.