噬菌体呈示单链抗体表达载体及小鼠非特异抗体库的构建  

Construction of ExFressing Vector for Phage Display scFv and a MouseUnspecified Antibody Library

作　　者：张卫国[1] 刘喜富[1] 王勇[1] 蒋欣[1] 林晴[1] 陈艾[1] 吴朝伟[1] 黄浩旻 李志华[1] 张屹 黄华梁[1] 

机构地区：[1]中国科学院遗传研究所,北京100101

出　　处：《Acta Genetica Sinica》1999年第2期99-106,共8页

基　　金：863计划资助

摘　　要：用大肠杆菌丝状噬菌体表面呈示技术构建抗体库的方法，为从抗原出发获得特异抗体提供了新的途径。报道的是，首先构建了一个用于噬菌体呈示抗体的噬粒表达载体ｐＦＵＷ８０，它具有既可以进行外分明表达，又可进行附着表达的特点。然后利用设计的一套扩增小鼠抗体重链和轻链可变区基因片段的ＰＣＲ引物，从未免疫小鼠脾细胞中扩增出了抗体重、轻链可变区基因，构建了一个１．２×１０６库容的小鼠非特异性单链抗体库。从这个抗体库中，筛选出了针对人ＩｇＧ的单链抗体噬菌体，并进行了ＥＬＩＳＡ检测和部分序列分析。这一初步结果为今后继续利用这一系统进行研究奠定了基础。he display of antibody gene library on the surface of E.coli filamentous phageoffers a new method of obtaining antibodies against specific antigens.In the paper,itis reported that a phagemid for phage display antibody,named after pFUW80,characterized expressing scFv either secretly of stickily.With a series of designed PCRprimers,heavy-chain and light-chain variable region genes of mice antibodies wereamplified and a mouse unspecified single-chain antibody library with the size of 1.2×106 clones was constructed.From this library,phage was selected out against antigenhuman lgG and detected by ELISA and partial sequence analysis.These primaryresults established the foundation for future research using this system.

关 键 词：抗体库技术 单链抗体 pFUW80 

分 类 号：R392.13[医药卫生—免疫学] Q78[医药卫生—基础医学]