Purification of retroviral vectors

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 7/04 (2006.01) C12N 7/02 (2006.01) C12N 15/86 (2006.01) C12N 15/867 (2006.01)

Patent

CA 2181066

A method of purifying retroviral vector particles which comprises culturing retroviral vector producer cells which are capable of generating retroviral vector particles, and obtaining a supernatant containing retroviral vector particles from the culture of retroviral vector producer cells. The supernatant is clarified, and then concentrated. The supernatant then is contacted with a precipitation agent, and the supernatant and precipitation agent are centrifuged. A precipitate is recovered, resuspended, and subjected to a high-speed centrifugation. Retroviral particles then are recovered. The above method provides for increased recovery of retroviral vector particles which may be employed in gene therapy.

L'invention concerne un procédé de purification de particules vectrices rétrovirales, consistant à mettre en culture des cellules productrices de vecteurs rétroviraux pouvant générer les particules vectrices rétrovirales, et à obtenir un supernageant contenant des particules vectrices rétrovirales à partir de la culture des cellules productrices de vecteurs rétroviraux. Le supernageant est ensuite clarifié, puis concentré. Le supernageant est alors mis en contact avec un agent de précipitation, et le supernageant est centrifugé avec l'agent de précipitation. Un précipitat est recueilli, remis en suspension et soumis à une centrifugation à haute vitesse. Les particules rétrovirales sont ensuite recueillies. Le procédé ci-dessus permet de recueillir, en quantités accrues, des particules vectrices rétrovirales qui peuvent être utilisées en thérapie génique.

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