Imidazole-free purification of His3-tagged recombinant proteins using ssDNA aptamer-based affinity chromatography

被引：15

作者：

Bartnicki, Filip ^{[1
]}

Kowalska, Ewa ^{[1
]}

Pels, Katarzyna ^{[1
]}

Strzalka, Wojciech ^{[1
]}

机构：

[1] Jagiellonian Univ, Fac Biochem Biophys & Biotechnol, Dept Plant Biotechnol, PL-30387 Krakow, Poland

来源：

JOURNAL OF CHROMATOGRAPHY A | 2015年 / 1418卷

关键词：

DNA aptamers; SELEX; Histidine tag; IMAC; Imidazole; CELL NUCLEAR ANTIGEN; IN-VITRO SELECTION; DNA-POLYMERASE; RNA MOLECULES; BACTERIOPHAGE-T4; EXPRESSION; PROTEASE; LIGANDS; VIRUS; YEAST;

D O I：

10.1016/j.chroma.2015.09.055

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Immobilized metal ion affinity chromatography (IMAC) is widely used for the purification of many different His(6)-tagged recombinant proteins. On the one hand, it is a powerful technique but on the other hand it has its disadvantages. In this report, we present the development of a unique ssDNA aptamer for the purification of His(3)-tagged recombinant proteins. Our study shows that stability of the His(3)-tag/H3T aptamer complex can be controlled by the sodium ion concentration. Based on this feature, we demonstrate that H3T aptamer resin was successfully employed for the purification of three out of four tested His(3)-tagged recombinant proteins from an E. coli total protein extract using imidazole-free buffers. Finally, we show that the purity of His(3)-tagged proteins is superior when purified with the help of the H3T aptamer in comparison with Ni-NTA resin. (C) 2015 Elsevier B.V. All rights reserved.

引用

页码：130 / 139

页数：10

共 35 条

[1] Upgrading SELEX Technology by Using Lambda Exonuclease Digestion for Single-Stranded DNA Generation [J].