Method for obtaining retroviral packaging cell lines...

C - Chemistry – Metallurgy – 12 – N

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C12N 5/10 (2006.01) C12N 15/86 (2006.01) C12N 15/867 (2006.01) A61K 48/00 (2006.01)

Patent

CA 2238436

This invention provides a method for obtaining a recombinant retroviral packaging cell capable of producing retroviral vectors as well as the recombinant packaging cell obtained by the method. Also provided is a method of producing recombinant retroviral particles obtained by introducing into the packaging cells obtained according to the methods disclosed herein, a recombinant retroviral vector and propagating the resulting producer cells under conditions favorable for the production and secretion of retroviral vector supernatant. The retroviral supernatants produced by these methods also is claimed herein. This invention further provides a method for screening retroviral vector supernatant for high transduction efficiency and methods for producing retroviral vector supernatant for transducing cells with high efficiency in gene therapy applications.

L'invention concerne un procédé permettant de produire une cellule d'encapsidation rétrovirale de recombinaison capable de produire des vecteurs rétroviraux ainsi que la cellule d'encapsidation de recombinaison produite selon ce procédé. Elle porte aussi sur un procédé de production de particules rétrovirales de recombinaison que l'on produit en introduisant dans les cellules d'encapsidation obtenues selon les procédés de l'invention, un vecteur rétroviral de recombinaison et en provoquant la propagation des cellules productrices résultantes dans des conditions favorables pour la production et la sécrétion de surnageant vectoriel rétroviral. Les surnageants rétroviraux produits selon ces procédés sont également décrits. L'invention se rapporte également à une méthode qui consiste à analyser l'efficacité de transduction élevée du surnageant vectoriel et à des procédés de production très efficaces de surnageant vectoriel rétroviral pour la transduction de cellules dans des applications de thérapie génique.

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