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当前位置: 首页 > 文章 > 抗克伦特罗单链抗体可溶性表达及特性鉴定 食品科学 2007,28 (12) 332-335
Position: Home > Articles > Soluble Expression and Characterization of Single Chain Fv Against Clenbuterol FOOD SCIENCE 2007,28 (12) 332-335

抗克伦特罗单链抗体可溶性表达及特性鉴定

作  者：
王弘;潘科;杨金易;张宏斌;王捷;武婕;雷红涛;孙远明
单  位：
广州军区广州总医院医学试验科;广东省教育厅食品质量安全重点实验室华南农业大学食品质量与安全研究所
关键词：
克伦特罗;单链抗体;可溶性表达
摘  要：
目的:表达抗克伦特罗(CBL)可溶性单链抗体(scFv)并进行抗原结合活性鉴定。方法:利用呈现scFv的重组噬菌体感染大肠杆菌HB2151,IPTG诱导可溶性scFv表达。渗透休克法提取菌体细胞周质腔中scFv。经SDS-PAGE、Western-Blotting以及间接ELISA法分析检测可溶性scFv的表达,并采用间接竞争ELISA法鉴定scFv的抗原结合活性。结果:细菌培养上清及周质腔中均有抗CBL可溶性scFv表达,其分子量约为29kD;上清液及周质腔中scFv效价分别为1:80,1:1600,能够被CBL竞争性抑制,IC50分别为0.78、0.95ng/ml。结论:可溶性抗CBLscFv在大肠杆菌HB2151中获得成功表达,表达的scFv与CBL具有很好的结合活性,可用于进一步建立CBL相应的免疫学检测方法。
译  名：
Soluble Expression and Characterization of Single Chain Fv Against Clenbuterol
作  者：
WANG Hong1,PAN Ke1,YANG Jing-yi1,ZHANG Hong-bin2,WANG Jie2,WU Jie2, LEI Hong-tao1,SUN Yuan-ming1,* (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)
关键词：
clenbuterol;single chain Fv;soluble expression
摘  要：
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.

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