Method of producing adenoviral vector stocks

C - Chemistry – Metallurgy – 12 – N

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C12N 7/00 (2006.01) C12N 5/00 (2006.01) C12N 15/861 (2006.01)

Patent

CA 2400746

The present invention provides a method of producing an adenoviral vector stock by providing a culture of cells permissive for growth of adenoviral vectors, wherein the cells are in a medium, culturing the culture under conditions to permit growth of the cells, perfusing fresh medium trough the culture for a period of about 1-6 hours, in an amount of at least about two times the volume of medium in the culture, while the density of the cells in the medium is about 40-70 % of the density of cells obtained in the medium when the growth of the culture is in the stationary phase, contacting the culture with adenoviral vectors under conditions permissive for the infection of the cells, culturing the infected cells to replicate the adenoviral vectors, and harvesting the infected cells to obtain an adenoviral vector stock. A method of obtaining a synchronous culture of cells is also provided.

La présente invention porte sur un procédé de production d'une population de vecteurs adénoviraux qui consiste à obtenir une culture de cellules autorisant la prolifération des vecteurs adénoviraux, ces cellules se trouvant dans un milieu, mettre en culture la culture dans des conditions autorisant la prolifération des cellules, perfuser un milieu frais dans la culture sur une durée d'environ 1 à 6 heures, la densité des cellules dans le milieu représentant environ entre 40 à 70 % de la densité des cellules obtenues dans le milieu lorsque la croissance de la culture est en phase stationnaire, mettre en contact la culture avec des vecteurs adénoviraux dans des conditions autorisant l'infection des cellules, mettre en culture les cellules infectées afin de répliquer les vecteurs adénoviraux et recueillir les cellules infectées pour obtenir une population de vecteurs adénoviraux. L'invention porte également sur un procédé d'obtention d'une culture synchrone de cellules.

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