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Specific recognition of supercoiled plasmid DNA in arginine affinity chromatography
Authors:Sousa F  Matos T  Prazeres D M F  Queiroz J A
Institution:a Centro de Investigação em Ciências da Saúde (CICS), Universidade da Beira Interior, 6201-001 Covilhã, Portugal
b Institute for Biotechnology and Bioengineering (IBB), Centro de Engenharia Biológica e Química, Instituto Superior Técnico, 1049-001 Lisboa, Portugal
Abstract:Arginine chromatography was used to fully separate supercoiled and open circular plasmid DNA (pDNA) isoforms. The results show that the arginine matrix promotes multiple interactions with pDNA, including not only electrostatic and hydrophobic but also biorecognition of nucleotide bases by the arginine ligand. The strong interactions occurring with DNA backbone provide stability, conducting to high effectiveness of arginine support to bind pDNA at low ionic strength. The specific interaction of arginine with sc pDNA could be due to the ability of arginine matrix to be involved in complex interactions that are partly dependent on the conformation of the DNA molecule.
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