Apparatus and method for preparative scale purification of...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/68 (2006.01) C07H 19/00 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C12P 19/34 (2006.01)

Patent

CA 2498518

Apparatus and methods are described for pharmaceutical grade manufacture extrachromosomal nucleic acids from cell lysates using flotation to separate and eliminate undesired insoluble cellular debris including chromosomal DNA from the lysates. A gas is introduced to controllably generate bubbles that reduce the density of the cell debris and create a buoyant flocculent phase that can be readily separated from, and thus provide, a substantially clarified fluid lysate phase that is enriched in extrachromosomal DNA but substantially depleted of cellular proteins and chromosomal DNA.

L'invention concerne un appareil et des procédés de fabrication de qualité pharmaceutique d'acides nucléiques extrachromosomiques à partir de lysats cellulaires utilisant la flottation pour séparer et éliminer les débris cellulaires insolubles indésirables, y compris l'ADN chromosomique provenant des lysats. On a produit un gaz pour générer de manière contrôlée des bulles réduisant la densité des débris cellulaires et pour créer une phase flottante de foculation prête à être séparée, et de ce fait obtenir, une phase de lysat fluide sensiblement clarifié enrichie en ADN extrachromosomique, mais sensiblement vidée de protéines cellulaires et d'ADN chromosomique.

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