title: Binding and elution strategy for improved performance of arginine affinity chromatography in supercoiled plasmid DNA purification.
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.
authors.affiliation: CICS, Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, 6201-001 Covilhã, Portugal.
authors.forename: F
authors.initials: F
authors.lastname: Sousa
authors.forename: D M F
authors.initials: DM
authors.lastname: Prazeres
authors.forename: J A
authors.initials: JA
authors.lastname: Queiroz
mesh_terms: D001120:Arginine / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D004278:DNA, Superhelical / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics; D010957:Plasmids; D012685:Sepharose / Q000737:chemistry; D012965:Sodium Chloride / Q000737:chemistry
chemical_list: D004278:DNA, Superhelical; D012965:Sodium Chloride; D012685:Sepharose; D001120:Arginine