Method of purification of recombinant viral vectors...

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 15/86 (2006.01) C07K 14/82 (2006.01) C12N 7/02 (2006.01) C12Q 1/00 (2006.01) A61K 48/00 (2006.01)

Patent

CA 2214837

The invention provides a method for purifying viral vectors containing therapeutic genes for use in gene therapy. The invention comprises a method of purification from a cell lysate of a recombinant viral vector containing a therapeutic gene, which comprises: a) treating said lysate with an enzymatic agent that selectively degrades both unencapsulated DNA and RNA; b) chromatographing the treated lysate from step a) on a first resin; and c) chromatographing the eluant from step b) on a second resin; wherein one resin is an anion exchange resin and the other is an immobilized metal ion chromatography (IMAC) resin.

Cette invention concerne un procédé de purification de vecteurs viraux contenant des gènes thérapeutiques utilisés en thérapie génétique. Cette invention consiste en un procédé de purification à partir d'un lysat cellulaire d'un vecteur viral recombinant contenant un gène thérapeutique, lequel procédé comporte les étapes suivantes: a) le lysat est traité par un agent enzymatique qui dégrade sélectivement l'ADN et l'ARN non encapsulés; b) le lysat traité en a) est ensuite chromatographié sur une première résine; c) l'éluant obtenu en b) est à son tour chromatographié sur une seconde résine. L'une des résines est une résine permettant l'échange d'anions tandis que la seconde est une résine de chromatographie d'ions métalliques immobilisés (IMAC).

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