抗克伦特罗噬菌体单链抗体库的构建、筛选及鉴定  

Construction,Screening and Identification of Phage scFv Library Against Clenbuterol

作　　者：潘科[1] 王弘[1] 张宏斌[2] 王捷[2] 雷红涛[1] 杨金易[1] 孙远明[1] 

机构地区：[1]华南农业大学食品质量安全广东省教育厅重点实验室,广东广州510640 [2]广州军区广州总医院医学实验中心,广东广州510010

出　　处：《华南理工大学学报（自然科学版）》2005年第11期51-54,共4页Journal of South China University of Technology(Natural Science Edition)

基　　金：国家自然科学基金资助项目(30400354;20477012);广东省自然科学基金资助项目(04300502)

摘　　要：利用噬菌体展示技术构建克伦特罗(CBL)单链抗体(scFv)库,从中筛选CBL特异性噬菌体scFv,从而成功扩增出抗CBL的VL,VH基因片段并采用重叠延伸PCR拼接为全长的scFv基因片段,抗体库库容约为1.6×104,经4轮吸附—洗脱—扩增的富集,采用酶联免疫吸附测定(ELISA)法筛选到6个具有CBL结合活性的噬菌体scFv,为进一步大量表达CBL单链抗体奠定了基础.Phage single-chain Fv （scFv） libraries with a size of about 1.6 x 104 were constructed for clenbuterol （CBL） by the phage display technology, and then, specific anti-CBL scFv was screened. Thus, the variable heavy （VH） and light （VL） anti-CBL genes were successfully amplified and scFv gene was assembled by overlapping PCR. After 4 times of enriched procedure in the order of adsorption-elution-amplification, 6 CBL-positive phage scFv clones were screened by ELISA. This research lays a foundation for the further expression of anti-CBL scFv.

关 键 词：克伦特罗 单链抗体 噬菌体展示 

分 类 号：R155[医药卫生—营养与食品卫生学]