Chromatographic purification of plasmid DNA for clinical applications (gene therapy)

J J Romanova; I I Salafutdinov; N M Zamalyutdinova; A A Rizvanov

doi:10.23868/gc121649

Genes & Cells ›› 2012, Vol. 7 ›› Issue (3) : 142 -145. DOI: 10.23868/gc121649

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Chromatographic purification of plasmid DNA for clinical applications (gene therapy)

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Abstract

For the successful application of plasmid vectors in genetherapy protocols it is necessary to develop methods forpurification of highly homogeneous preparations of recombinantDNA that do not contain contaminants, primarily chromosomalDNA, bacterial proteins, RNA and endotoxins. In the courseof our study we performed optimization of the purification ofplasmid supercoiled DNA by three chromatographic steps froman alkaline lysate of bacterial strain of E. coli. We determinedan optimal conditions for alkaline lysis step in order to increasethe yield and minimize the duration of the plasmid purificationby gel filtration.

Keywords

recombinant DNA plasmid / gel filtration / anion exchange chromatography / salting-out chromatography / gene therapy

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J J Romanova, I I Salafutdinov, N M Zamalyutdinova, A A Rizvanov. Chromatographic purification of plasmid DNA for clinical applications (gene therapy). Genes & Cells, 2012, 7(3): 142-145 DOI:10.23868/gc121649

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