Process for producing endotoxin-free or endotoxin-poor...

Details Process for producing endotoxin-free or endotoxin-poor... Process for producing endotoxin-free or endotoxin-poor...

C - Chemistry, Metallurgy

07

H

C07H 21/00 (2006.01) A61K 31/70 (2006.01) A61K 48/00 (2006.01) C12N 15/10 (2006.01) C12N 15/87 (2006.01)

Patent

CA 2182388

A process for the isolation and purification of nucleic acids and/or oligonucleotides for use in gene therapy wherein said nucleic acids and/or oligonucleotides are isolated or purified from an essentially biological source, characterized in that said essentially biological sources are lysed, the frac-tions obtained are optionally freed or depleted from the remainder of said biological sources by per se known mechanical methods, such as centrifugation, filtration; - the fractions thus treated are subsequently treated with affinity chromatographic material or with inorganic chromatographic material for the removal of endotoxins; followed by - isolation of said nucleic acids and/or oligonucleotides on an anion exchanger which is designed such that DNA begins to desorb from the anion exchanger only at an ionic strength corresponding to a sodium chloride solution of a concentration higher by at least 100 mM than one corre-sponding to the ionic strength at which RNA begins to desorb from the anion exchanger material.

Un procédé permet d'isoler et de purifier des acides nucléiques et/ou des oligonucléotides utiles en thérapie génique. Les acides nucléiques et/ou les oligonucléotides sont isolés ou purifiés à partir d'une source essentiellement biologique. Le procédé se caractérise en ce que les sources essentiellement biologiques sont désagrégées, les résidus de sources biologiques sont, le cas échéant, éliminés ou réduits dans les fractions ainsi obtenues, par un procédé mécanique connu en soi, tel que la centrifugation ou le filtrage. Les fractions ainsi traitées sont ensuite traitées avec des matériaux de chromatographie par affinité ou de chromatographie inorganique afin d'éliminer les endotoxines, puis les acides nucléiques et/ou les oligonucléotides sont isolés sur un échangeur d'anions conçu de sorte que l'ADN ne commence à être désorbé par l'échangeur d'anions que lorsque la force ionique d'une solution de chlorure de sodium dépasse d'au moins 100 mM environ la force ionique à laquelle l'ARN du matériau d'échange d'anions commence à être désorbée par l'échangeur d'anions.

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