FOOD SCIENCE ›› 2007, Vol. 28 ›› Issue (12): 332-335.

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Soluble Expression and Characterization of Single Chain Fv Against Clenbuterol

WANG  Hong, PAN  Ke, YANG  Jin-Yi, ZHANG  Hong-Bin, WANG  Jie, WU  Jie, LEI  Hong-Tao, SUN  Yuan-Ming   

  1. 1. Key Laboratory of Food Quality and Safety, Department of Education of Guangdong Province, Research Institute of Food Quality and Safety, South China Agricultural University, Guangzhou 510640, China; 2.Department of Medical Research, Guangzhou Army General Hospital, Guangzhou 510010, China
  • Online:2007-12-15 Published:2011-11-22

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Abstract: Objective: To express and characterize the soluble single chain Fv (scFv) against clenbuterol (CBL). Methods: The soluble anti-CBL scFv was expressed by E.coli. HB2151 infected by recombinant phages after being induced by IPTG and extracted from the periplasmic through osmotic shock. The characterization and antigen-binding activity of the scFv were identified by SDS-PAGE, Western-Blotting, and indirected by ELISA and competitive ELISA. Results: The scFv with MW 29kD is expressed both in supernatant and periplasmic. The titer of scFv in supernatant is 1:160 and that in periplasmic extracts 1:3200. The indirect competitive ELISA showed that the scFv in both supernatant and periplasmic can competitively combine with CBL, whereas IC50 is 0.78 ng/ml and 0.95 ng/ml respectively. Conclusion: The soluble scFv directed against CBL is expressed successfully and shows the ideal binding activity to CBL. The results showed that the corresponding immnunoassay methods can be set up by this scFv.

Key words: clenbuterol, single chain Fv, soluble expression