Single-step Purification of Molecular Chaperone GroEL by Expanded Bed Chromatography

Abstract

Abstract: Expanded bed adsorption (EBA) is an integrative downstream processing technique for the
purificationof biological substances directly from unclarified feedstock. In this study,
molecular chaperone GroEL, an importantprotein folding helper both in vivo and in vitro,
was purified by the single-step EBA technique from the unclarifiedhomogenate of recombinant
E. coli cells. Compared with packed bed adsorption, the EBA technique provideda single-step
approach to yield an electrophoretic purity of GroEL. After the homogenate loading and
columnwashing in the expanded bed mode, the GroEL protein was recovered by stepwise salt-
gradient elution in packed-bed or expanded-bed modes, respectively. The expanded-bed
elution mode was found as efficient as the packed-bedmode in the purification of GroEL from
cell disruptate.

Key words: expanded bed adsorption, molecular chaperone, GroEL, purification

摘要： Expanded bed adsorption (EBA) is an integrative downstream processing technique for the
purificationof biological substances directly from unclarified feedstock. In this study,
molecular chaperone GroEL, an importantprotein folding helper both in vivo and in vitro,
was purified by the single-step EBA technique from the unclarifiedhomogenate of recombinant
E. coli cells. Compared with packed bed adsorption, the EBA technique provideda single-step
approach to yield an electrophoretic purity of GroEL. After the homogenate loading and
columnwashing in the expanded bed mode, the GroEL protein was recovered by stepwise salt-
gradient elution in packed-bed or expanded-bed modes, respectively. The expanded-bed
elution mode was found as efficient as the packed-bedmode in the purification of GroEL from
cell disruptate.

关键词: expanded bed adsorption;molecular chaperone;GroEL;purification

TONG Xiaodong, YANG Zheng, DONG Xiaoyan, SUN Yan. Single-step Purification of Molecular Chaperone GroEL by Expanded Bed Chromatography[J]. .

佟晓冬, 杨征, 董晓燕, 孙彦. 利用膨胀床吸附技术单步纯化分子伴侣—GroEL[J]. CIESC Journal.

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