Method for purifying plasmid dna

C - Chemistry – Metallurgy – 12 – N

Patent

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C12N 1/06 (2006.01) C12N 15/10 (2006.01) C12Q 1/68 (2006.01)

Patent

CA 2559368

This invention provides a process for the continuous alkaline lysis of a bacterial suspension in order to harvest pDNA. It further provides for optional additional purification steps, including lysate filtration, anion exchange chromatography, triplex affinity chromatography, and hydrophobic interaction chromatography. These optional purification steps can be combined with the continuous lysis in order to produce a highly purified pDNA product substantially free of gDNA, RNA, protein, endotoxin, and other contaminants.

L'invention concerne un procédé de lyse alcaline continue d'une suspension bactérienne en vue de cultiver de l'ADNp. Elle concerne également des étapes de purification supplémentaires facultatives, notamment la filtration de lysat, la chromatographie échangeuse d'anions, la chromatographie par affinité de triplex et la chromatographie hydrophobe. Ces étapes de purification facultatives peuvent être combinées à la lyse continue en vue de produire un produit d'ADNp hautement purifié sensiblement dépourvu d'ADNg, d'ARN, de protéines, d'endotoxines, et d'autres contaminants.

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