Prokaryotic Expression and Purification of Single-chain Variable Fragment（scFv）Against Staphylococcus aureus

doi:10.13560/j.cnki.biotech.bull.1985.2016.11.022

Abstract

Abstract: This work aims to clone and express single-chain variable region fragment（scFv）of Staphylococcus aureus in Escherichia coli,and to obtain the target protein with scFv activity. A plasmid containing target scFv gene was constructed,then transformed into a prokaryotic expression strain to be induced,and the activity of the harvested protein was identified. As results：（1）Recombinant plasmid pCold I-scFv was successfully constructed,and transformed into the expression strain of Escherichia coli.（2）After induction,the target proteins mainly exist in pellet in the form of inclusion bodies. （3）Inclusion body was dissolved successfully using 4 mol/L urea. （4）Purified and renatured target protein by column chromatography and dialysis was favorable. （5）The refolded protein showed specific binding activity with S. aureus in ELISA experiment. Conclusively,the target protein with S. aureus antibody activity was successfully acquired by plasmid construction and prokaryotic expression,inclusion body dissolving and refolding.

Key words: soluble expression, inclusion, protein purification, protein refolding, active protein

LI Jing-quan, XU Yong-ping, WANG Xi-tao, LI Yuan, WANG Li-li, LI Xiao-yu. Prokaryotic Expression and Purification of Single-chain Variable Fragment（scFv）Against Staphylococcus aureus[J]. Biotechnology Bulletin, 2016, 32(11): 194-201.

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