Journal
BIOMEDICAL CHROMATOGRAPHY
Volume 23, Issue 2, Pages 160-165Publisher
WILEY
DOI: 10.1002/bmc.1097
Keywords
affinity chromatography; arginine-agarose; chromatographic retention; plasmid purification
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Funding
- Portuguese Foundation for Science and Technology [PTDC/EQU-EQU/65492/2006, SFRH/ BD/24184/2005]
- Fundação para a Ciência e a Tecnologia [PTDC/EQU-EQU/65492/2006, SFRH/BD/24184/2005] Funding Source: FCT
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New interesting strategies for plasmid DNA (pDNA) purification were designed, exploiting affinity interactions between amino acids and nucleic acids. The potential application of arginine-based chromatography to purify pDNA has been recently described in our work, however, to achieve higher efficiency and selectivity in arginine affinity chromatography, it is essential to characterize the behaviour of binding/elution of supercoiled (sc) isoforms. In this study, two different strategies based on increased sodium chloride (225-250 mM) or arginine (20-70 mM) stepwise gradients are described to purify sc isoforms. Thus, it was proved that well-defined binding/elution conditions are crucial to enhance the purification performance, resulting in an improvement of the final plasmids yields and transfection efficiency, as this could represent a significant impact on therapeutic applications of the purified sc isoform. Copyright (c) 2008 John Wiley & Sons, Ltd.
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