Expression of functional single-chain variable domain fragment antibody (scFv) against mycotoxin zearalenone in Pichia pastoris

被引:0
作者
Hyun-Joo Chang
Sung-Wook Choi
Hyang Sook Chun
机构
[1] Korea Food Research Institute,
来源
Biotechnology Letters | 2008年 / 30卷
关键词
Codon-optimization; ScFv; Surface plasmon resonance; Zearalenone;
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学科分类号
摘要
A synthetic gene coding for single-chain variable domain fragment antibody against mycotoxin zearalenone (scFv-ZEN) has been designed, constructed and expressed in Pichia pastoris. The native scFv-ZEN sequence was optimized to Pichia preference codon usage. The expression level of codon-optimized scFv-ZEN was slightly higher than that of native scFv-ZEN, and its maximum yield reached 328 mg total protein/l in flask culture. The binding activities of two selected clones to ZEN using surface plasmon resonance analysis were comparable or better than that of monoclonal antibody. Our results demonstrate the potential of soluble scFv-ZEN for developing a rapid and affordable immunoassay for detection of ZEN in food and feedstuff.
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页码:1801 / 1806
页数:5
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共 74 条
[1]  
Backmann N(2005)A label-free immunosensor array using single-chain antibody fragments Proc Natl Acad Sci USA 102 14587-14592
[2]  
Zahnd C(2000)Fermentation, purification, and efficacy of a recombinant vaccine candidate against Protein Expr Purif 18 327-337
[3]  
Huber F(2006) neurotoxin type F from J Agric Food Chem 54 815-822
[4]  
Bietsch A(2007)Codon optimization of J Agric Food Chem 55 8353-8358
[5]  
Plückthun A(2004)1 gene for improving expression in Protein Expr Purif 37 18-26
[6]  
Lang H-P(2006) and biochemical characterization of the purified recombinant LIP1 lipase Protein Expr Purif 47 249-257
[7]  
Güntherodt H-J(2005)Stable isotope dilution analysis of the J Biotechnol 120 38-45
[8]  
Hegner M(2002) mycotoxin zearalenone Microchem J 72 315-321
[9]  
Gerber C(1999)An optimized fermentation process for high-level production of a single-chain Fv antibody fragment in Biotechnol Lett 21 561-567
[10]  
Byrne MP(1995)Codon optimization, expression, and characterization of an internalizing anti-ErbB2 single-chain antibody in Food Technol 49 120-128