Expression and purification of human perforin by sf9 insect cells

Basic & Clinical Medicine ›› 2020, Vol. 40 ›› Issue (1): 54-59.

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Expression and purification of human perforin by sf9 insect cells

CHEN Feng-ye, LIU Meng-yu, ZHOU Ya-bo, LIANG Xiao-yu, HUANG Bo, XIE Jing^*

National Key Laboratory of Medical Molecular Biology, Department of Immolunology, Institute of Basic Medical Sciences CAMS & School of Basic Medicine PUMC, Beijing 100005, China

Received:2019-10-12 Revised:2019-11-22 Online:2020-01-05 Published:2019-12-27
Contact: ^*xiejing@ibms.pumc.edu.cn

Abstract

Abstract: Objective To optimize the purification method of human perforin. Methods Perforin with His tag was expressed by sf9 insect cells, and then protein affinity chromatography was performed by Ni column device. The physicochemical property and biological activity of the purified protein was analyzed by coomassie brilliant blue staining, silver staining and flow cytometry. Results This method was more convenient and economical than traditional protein purification methods like ion exchange chromatography, molecular sieve and hydrophobic interaction chromatography, which providing new ideas for further research of perforin and purification of other protein. Conclusions Perforin can be expressed by sf9 insect cells and is purified by a simple method with biological activity.

Key words: protein purification, perforin, protein activity, pore-formation

CLC Number:

CHEN Feng-ye, LIU Meng-yu, ZHOU Ya-bo, LIANG Xiao-yu, HUANG Bo, XIE Jing. Expression and purification of human perforin by sf9 insect cells[J]. Basic & Clinical Medicine, 2020, 40(1): 54-59.

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Expression and purification of human perforin by sf9 insect cells

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