抗A型肉毒毒素人源单链抗体融合蛋白的重组设计  

Recombinant Design of Fc Fusion Protein of Human ScFv Against BoNTa

作　　者：王慧[1] 史晶[1] 孟露露[1] 李佩真[1] 白瑜[1] 荫俊[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100071

出　　处：《微生物学通报》2005年第4期11-14,共4页Microbiology China

基　　金：国家重点基础研究973项目资助(No.2002CB513205);十五总后指令性课题(No.IL033)

摘　　要：以获得的抗A型肉毒毒素单链抗体为模板,进行融合改构,将人IgG1的Fc片段连接到ScFv的C端,在大肠杆菌中实现抗体融合蛋白ScFv-Fc的表达,表达量30%以上,蛋白以包含体形式存在,经过体外变复性的抗体融合蛋白ScFv-Fc,进行Protein G Sepharose柱亲和层析纯化,纯度达90%～95%.体外活性检测结果表明,重组抗体融合蛋白ScFv-Fc可以特异结合A型肉毒类毒素抗原,其相对亲和力近似于母本单链抗体,其稳定性高于母本单链抗体.Human ScFv against botulinum neurotoxin serotype A （BoNTa） was modified by fusing human IgGl Fc to C terminal of ScFv. ScFv-Fc fusion protein was expressed at high level over 30% of total host cell proteins in E. coli Recombinant protein existed in inclusion body form. Renatured ScFv-Fc was purified to 90% - 95% by Protein G Sepharose column. In vitro ScFv-Fc could bind specific to toxiod BoNTa in ELISA. Recombinant ScFv-Fc had similar relative affinity to parent ScFv and had improved stability.

关 键 词：A型肉毒毒素 单链抗体融合蛋白 相对亲和力 稳定性 

分 类 号：R392.1[医药卫生—免疫学]