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Purification of supercoiled plasmid DNA using chromatographic processes

✍ Scribed by G. N. M. Ferreira; J. M. S. Cabral; D. M. F. Prazeres

Publisher: John Wiley and Sons
Year: 1998
Tongue: English
Weight: 76 KB
Volume: 11
Category: Article
ISSN: 0952-3499
DOI: 10.1002/(sici)1099-1352(199812)11:1/6<250::aid-jmr433>3.0.co;2-1

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✦ Synopsis

The interest in purifying injectable-grade plasmid DNA has increased with the development of gene therapy and DNA vaccination technologies. In this paper we develop a method for purifying a 4.8 kb plasmid based on chromatographic processes. An NaCl gradient was optimized on a Q Sepharose 1 column and plasmid was eluted at 800-820 mM NaCl in a broad peak. Supercoiled plasmid was isolated after a final Sepharcryl S1000 SF 1 gel filtration step. Final plasmid preparation was depleted of proteins and RNA, as revealed by the BCA assay and 1% agarose gel electrophoresis.

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