人源抗A型肉毒毒素单链抗体的体外亲和筛选  

Affinity Selection of Human ScFv Against Botulinum Neurotoxin Serotype A

作　　者：王慧[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1期22-25,共4页Microbiology China

基　　金：国家重点基础研究973项目资助(No.2002CB513205).

摘　　要：以重组制备的A型肉毒毒素保护性抗原为配体,对人源噬菌体免疫抗体文库进行体外定向亲和筛选,获得特异结合子,其中与抗原高亲和力结合的抗体克隆B17基因全长750bp,可编码250个氨基酸,抗体可变区基因同源分析表明,分属VH4和κchainⅡ家族,是一株人源特异单链抗体基因。人源单链抗体B17在大肠杆菌中获得了重组表达,表达产物可以竞争特异肉毒抗毒素马血清与抗原的结合,是国内首次获得的抗A型肉毒毒素保护性抗原的人源单链抗体,可以在肉毒毒素检测和治疗研究中发挥作用。In this work, rare specific binders to botulinum neurotoxin antigen were selected from human phage immunized library through three rounds of panning in vitro. Of them, clone ScFvB17 is of high affinity and specificity to target antigen. B17 is of 750bp encoding 250 amino acid. Analysis results of gene structure showed V-genes of B17 belonging respectively to VH4 orκ chainⅡ family are new antibody Fv genes. More important result showed that recombinant ScFvB17 expressed in E.coli could bind specific for neurotoxin competing with antiserum. Successful selection of human phage antibody library and obtaining specific ScFv against botulinum neurotoxin serotype A provide useful materials for further study on detection of neurotoxin and human engineering therapeutic antitoxin.

关 键 词：A型肉毒毒素 噬菌体文库 人源单链抗体 表达 

分 类 号：R392.1[医药卫生—免疫学]