Method of preparation of pharmaceutically grade plasmid dna

Details Method of preparation of pharmaceutically grade plasmid dna Method of preparation of pharmaceutically grade plasmid dna

C - Chemistry, Metallurgy

12

N

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

Patent

CA 2536931

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 aux fins de prélèvement d'ADNp. L'invention concerne également des étapes de purification supplémentaires facultatives, notamment une filtration de lysat, une chromatographie échangeuse d'anions, une chromatographie par affinité de triplex et une chromatographie par interaction hydrophobe. Ces étapes de purification facultatives peuvent être combinées avec la lyse continue, afin de produire un produit d'ADNp très purifié et sensiblement exempt d'ADNg, d'ARN, de protéine, d'endotoxine et d'autre contaminants.

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