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Dynamic binding capacity and specificity of 3, 8-diamino-6-phenylphenanthridine-Sepharose support for purification of supercoiled plasmid DNA.

J Chromatogr A.. 2013-07; 
C Caramelo-Nunes, D Bicho, P Almeida, JC Marco, C.T. Tomaz. CICS-UBI-CHealth Sciences Research Centre, University of Beira Interior, Av. Infante D. Henrique, 6200-506 Covilhã, Portugal
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摘要

Affinity chromatography represents a sole technique in purification of different biomolecules. The specific recognition between affinity ligands and target biomolecules has a major role in the specificity of the process. Therefore, choosing the right ligand is a crucial step for the development of a successful purification system. This work describes the application of the DNA intercalator 3,8 - diamino - 6 - phenylphenanthridine (DAPP) as a chromatographic affinity ligand for the specific separation and purification of supercoiled plasmid DNA (pDNA). The support was prepared by coupling DAPP onto an epoxy-activated Sepharose matrix, using mild conditions and resulting in a ligand density of 0.15 mmol DAPP/g de... More

关键词

DAPP-Sepharose; Affinity Chromatography; dissociation constant; dynamic binding capacity; supercoiled pDNA purification.
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