Biorecognition of supercoiled plasmid DNA isoform in lysine-affinity chromatography

被引:27
|
作者
Sousa, A. [1 ]
Sousa, F. [1 ]
Queiroz, J. A. [1 ]
机构
[1] Univ Beira Interior, CICS, P-6201001 Covilha, Portugal
关键词
Affinity chromatography; Lysine support; Supercoiled plasmid DNA; PURIFICATION; SEPARATION; AGAROSE;
D O I
10.1016/j.jchromb.2009.08.021
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The use of plasmid DNA-based therapeutics relies on procedures that efficiently purify the supercoiled plasmid isoform. The present study describes a new strategy that uses a lysine ligand in affinity chromatography to efficiently separate supercoiled and open circular plasmid DNA isoforms. To achieve higher specificity in this chromatography it is essential to characterize the behaviour of binding/elution of supercoiled isoforms. The results show that the lysine support promotes complex interactions with supercoiled isoform, according to ionic strength and temperature manipulation. (C) 2009 Elsevier B.V. All rights reserved.
引用
收藏
页码:3257 / 3260
页数:4
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