Nucleic acid affinity of clustered-charge anion exchange adsorbents: Effects of ionic strength and ligand density

被引:12
作者
Chen, Wen-Hsiang [1 ]
Fu, Joseph Y. [1 ]
Kourentzi, Katerina [1 ]
Willson, Richard C. [1 ,2 ]
机构
[1] Univ Houston, Dept Chem Engn, Houston, TX 77204 USA
[2] Univ Houston, Dept Biol & Biochem, Houston, TX 77204 USA
关键词
Ion-exchange chromatography; Clustered charge; Nucleic acids; Adsorption; Ligand density; SUPERCOILED PLASMID DNA; THERMODYNAMIC DRIVING FORCES; TAR RNA; CYTOCHROME B(5); COMPACTION AGENTS; CHROMATOGRAPHY; ARGININE; PURIFICATION; SELECTIVITY; BINDING;
D O I
10.1016/j.chroma.2010.11.024
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In previous work we demonstrated the improved protein-binding capacity and selectivity of ion-exchange adsorbents displaying a "clustered" rather than random, distribution of surface charges. For example, anion-exchange adsorbents displaying 5 mM of positive charge in the form of 1 mM penta-argininamide show much higher affinity and capacity for alpha-lactalbumin than do adsorbents displaying the same 5 mM total charge in the form of single dispersed argininamide charges. We also found that clustered adsorbents selectively favor proteins with inherent charge clustering. In the present work. "clustered" penta-argininamide adsorbents showed DNA binding capacity comparable to that of conventional dispersed adsorbents with 10-100-fold higher ligand density. We also observed that at moderate ionic strength the DNA affinity of all adsorbents tested increased with salt while RNA affinity decreased, so that selectivity for DNA over RNA was enhanced as salt concentration increased. (C) 2010 Elsevier B.V. All rights reserved.
引用
收藏
页码:258 / 262
页数:5
相关论文
共 31 条
[1]   Metal chelate affinity precipitation of RNA and purification of plasmid DNA [J].
Balan, S ;
Murphy, J ;
Galaev, I ;
Kumar, A ;
Fox, GE ;
Mattiasson, B ;
Willson, RC .
BIOTECHNOLOGY LETTERS, 2003, 25 (13) :1111-1116
[2]   SEPARATION OF NEUTRAL PROTEINS ON ION-EXCHANGE RESINS [J].
BOARDMAN, NK ;
PARTRIDGE, SM .
BIOCHEMICAL JOURNAL, 1955, 59 (04) :543-552
[3]   STERIC MASS-ACTION ION-EXCHANGE - DISPLACEMENT PROFILES AND INDUCED SALT GRADIENTS [J].
BROOKS, CA ;
CRAMER, SM .
AICHE JOURNAL, 1992, 38 (12) :1969-1978
[4]  
CALISKAN G, 2005, PHYS REV LETT, V95, P4
[5]  
Clifford D., 1983, REACT POLYM, V1, P77, DOI [DOI 10.1016/0167-6989(83)90040-5, 10.1016/0167-6989(83)90040-5]
[6]   Base flexibility in HIV-2 TAR RNA mapped by solution 15N,13C NMR relaxation [J].
Dayie, KT ;
Brodsky, AS ;
Williamson, JR .
JOURNAL OF MOLECULAR BIOLOGY, 2002, 317 (02) :263-278
[7]   Enhanced protein affinity and selectivity of clustered-charge anion-exchange adsorbents [J].
Fu, Joseph V. ;
Balan, Sindhu ;
Potty, Ajish ;
Nguyen, Van ;
Willson, Richard C. .
ANALYTICAL CHEMISTRY, 2007, 79 (23) :9060-9065
[8]   GIBBS FREE-ENERGY OF ADSORPTION FOR BIOMOLECULES IN ION-EXCHANGE SYSTEMS [J].
GERSTNER, JA ;
BELL, JA ;
CRAMER, SM .
BIOPHYSICAL CHEMISTRY, 1994, 52 (02) :97-106
[9]   PRESENCE OF A PREFERRED ANION-EXCHANGE BINDING-SITE ON CYTOCHROME B(5) - STRUCTURAL AND THERMODYNAMIC CONSIDERATIONS [J].
GILL, DS ;
ROUSH, DJ ;
WILLSON, RC .
JOURNAL OF CHROMATOGRAPHY A, 1994, 684 (01) :55-63
[10]   ADSORPTION HETEROGENEITY AND THERMODYNAMIC DRIVING FORCES IN ANION-EXCHANGE EQUILIBRIA OF CYTOCHROME-B(5) [J].
GILL, DS ;
ROUSH, DJ ;
WILLSON, RC .
JOURNAL OF COLLOID AND INTERFACE SCIENCE, 1994, 167 (01) :1-7