Purification and/or concentration of dna by cross-flow...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/10 (2006.01) B01D 61/00 (2006.01) C07H 1/08 (2006.01)

Patent

CA 2277165

The invention concerns a method of purifying and/or concentrating nucleic acids in a solution, the solution containing nucleic acid being guided tangentially past one or a plurality of semipermable membranes, such that the nucleic acid molecules are retained by the membranes and substances having a lower molecular weight can pass through the membranes and/or be adsorbed thereat, so that a purified and/or concentrated nucleic acid solution is obtained. The same method is carried out to separate endotoxins from a nucleic acid preparation. The invention further concerns the use of a cross-flow filtration system for purifying and/or concentrating nucleic acids in a solution and for separating endotoxins from a nucleic acid preparation. The invention also concerns the use of the nucleic acids purified and/or concentrated by the cross-flow filtration system for cloning, transformation, transfection and microinjection into cells, for use in gene therapy processes, DNA vaccination and/or for polymerase chain reaction (PCR).

L'invention concerne un procédé de purification et/ou de concentration d'acides nucléiques dans une solution, selon lequel la solution contenant l'acide nucléique est guidée de manière tangentielle pour passer sur une ou plusieurs membranes semi-perméables, de manière que les molécules d'acide nucléique soient retenues par les membranes et que des substances présentant un poids moléculaire moins important puissent passer à travers les membranes et/ou soient adsorbées au niveau de la membrane. On obtient ainsi une solution d'acide nucléique purifiée et/ou concentrée. On procède de la même manière pour séparer des endotoxines contenues dans une préparation d'acide nucléique. L'invention concerne en outre l'utilisation d'un système de filtration tangentielle pour purifier et/ou concentrer des acides nucléiques dans une solution, ainsi que pour séparer des endotoxines contenues dans une préparation d'acide nucléique. L'invention concerne par ailleurs l'utilisation des acides nucléiques purifiés ou concentrés par filtration tangentielle, à des fins de clonage, de transformation, de transfection, de micro-injection dans des cellules, d'application dans des procédés de thérapie génique, de vaccination par ADN et/ou de réaction en chaîne de la polymérase (PCR).

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