C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/11 (2006.01) A61K 31/711 (2006.01) A61K 39/00 (2006.01) A61K 48/00 (2006.01) C07H 21/04 (2006.01) C12N 15/10 (2006.01)
Patent
CA 2428885
The invention relates to a method of separating extra-chromosomal DNA from RNA. It also relates to DNA produced by the method and pharmaceuticals derived from such DNA, for example, DNA vaccines. The method comprises separating extra-chromosomal DNA from RNA without first digesting the RNA, and comprises: i) lysing cells comprising extra-chromosomal DNA to form a lysate; ii) clarifying the lysate; and iii) subjecting the clarified lysate to tangential flow ultra filtration under conditions, which allow a substantial amount of the RNA to pass through a membrane whilst the extra-chromosomal DNA is retained. The preferred conditions include the use of a diafiltration buffer of low conductivity and diafiltration volumes of over 10 volume equivalents.
L'invention concerne un procédé permettant de séparer l'ADN extrachromosomique de l'ARN. L'invention concerne également de l'ADN produit par ce procédé, ainsi que des produits pharmaceutiques dérivés de cet ADN, par exemple des vaccins par ADN. Ce procédé consiste à séparer l'ADN extrachromosomique de l'ARN sans digestion préalable de l'ARN et consiste à : i) lyser des cellules contenant l'ADN extrachromosomique pour former un lysat, ii) clarifier le lysat et iii) soumettre le lysat clarifié à une ultrafiltration tangentielle dans des conditions telles qu'une importante quantité de l'ARN passe à travers une membrane tandis que l'ADN extrachromosomique est retenu. Les conditions préférées comprennent l'utilisation d'un tampon de diafiltration de faible conductivité et de volumes de diafiltration supérieurs à 10 équivalents de volume.
Charlton Henry
Eon-Duval Alex
Macduff Robert
Glaxo Group Limited
Ogilvy Renault Llp/s.e.n.c.r.l.,s.r.l.
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