Method for preparing a plasmid dna

C - Chemistry – Metallurgy – 12 – N

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C12N 15/10 (2006.01) A61K 31/70 (2006.01) A61K 48/00 (2006.01) C07H 1/06 (2006.01) C07H 21/00 (2006.01) C07K 14/47 (2006.01) A61K 38/00 (2006.01)

Patent

CA 2266123

The invention concerns a method for preparing a plasmid DNA using a wet cell biomass comprising after resuspension of said biomass the following steps:, alkaline lysis, high ionic strength acidification, elimination of solubles, endotoxin and contaminating RNA reduction, filtering gel chromatography and conditioning. The invention also concerns a pharmaceutical composition containing a plasmid DNA and its use for gene therapy.

La présente invention concerne un procédé de préparation d'un ADN plasmidique à partir d'une biomasse cellulaire humide comprenant après la resuspension de ladite biomasse, une étape de lyse alcaline, une étape d'acidification à force ionique élevée, une étape d'élimination des insolubles, une étape de réduction des endotoxines et des ARN contaminants, une étape de chromatographie de gel de filtration et une étape de conditionnement. Elle a également pour objet une composition pharmaceutique comprenant un ADN plasmidique ainsi préparé et son utilisation à des fins de thérapie génique.

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