Binding and elution strategy for improved performance of arginine affinity chromatography in supercoiled plasmid DNA purification |
| |
Authors: | Sousa F Prazeres D M F Queiroz J A |
| |
Affiliation: | 1. CICS, Centro de Investiga??o em Ciências da Saúde, Universidade da Beira Interior, 6201‐001 Covilh?, Portugal;2. IBB, Institute for Biotechnology and Bioengineering, Centro de Engenharia Biológica e Química, Instituto Superior Técnico, 1049‐001 Lisboa, Portugal |
| |
Abstract: | 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. |
| |
Keywords: | affinity chromatography arginine–agarose chromatographic retention plasmid purification |
本文献已被 PubMed 等数据库收录! |
|