Ph dependent ion exchange matrix and method of use in the...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/10 (2006.01) B01J 39/26 (2006.01) B01J 41/20 (2006.01) C07H 21/00 (2006.01)

Patent

CA 2372054

pH dependent ion exchange matrices are provided, with methods for making such matrices, and methods for using such matrices to isolate a target nucleic acid, as such as plasmid DNA, chromosomal DNA, or RNA from contaminants, including proteins, lipids, cellular debris, or other nucleic acids. Each pH dependent ion exchange matrix of this invention comprises at least two different ion exchange functional groups, one of which is capable of acting as an anion exchanger at a first pH, and the other of which is capable of acting as a cation exchanger at a second, higher pH. The matrix has an overall neutral charge in a pH range between the first and second pH. The pH dependent ion exchange matrices of the present invention are designed to bind to the target nucleic acid at a pH wherein the overall charge of the matrix is positive, and to release the target nucleic acid as the pH of the surrounding solution is increased. The target nucleic acid can be released from the pH dependent matrix in little or no salt and at about a neutral pH. The matrices and methods of this invention enable one to isolate a target nucleic acid in very few steps, without the use of hazardous chemicals. Target nucleic acids isolated using the pH dependent ion exchange matrices according to the present invention can be used immediately without further extraction or isolation.

Cette invention concerne des matrices échangeuses d'ions dépendant du pH, leurs procédés de fabrication et des méthodes d'utilisation desdites matrices pour isoler un acide nucléique cible tel qu'un ADN plasmide, un ADN chromosomique ou un ARN de substances contaminantes dont des protéines, des lipides, des débris cellulaires ou d'autres acides nucléiques. Chacune des matrice échangeuses d'ions dépendant du pH selon l'invention comprend au moins deux groupes fonctionnels différents dont l'un est capable d'agir comme échangeur d'anions à un premier pH, l'autre comme échangeur de cations à un second pH, plus élevé. Entre le premier et le second pH, la matrice a une charge globale neutre. Les matrices échangeuses d'ions dépendant du pH selon l'invention sont conçues pour fixer l'acide nucléique cible à un niveau de pH où la charge globale de la matrice est positive, et pour libérer ce même acide au fur et à mesure que le pH de la solution environnante augmente. L'acide nucléique cible peut être libéré de la matrice dépendant du pH avec peu ou pas de sel et pour un pH pratiquement neutre. Grâce aux matrices et aux méthodes selon l'invention, il est possible d'isoler un acide nucléique cible en quelques opérations, sans recours à des produits chimiques dangereux. Les acides nucléiques isoles au moyen de matrices échangeuses d'ions dépendant du pH peuvent être utilisés immédiatement, sans autre opération d'extraction ou d'isolement.

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