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.
基金
863计划资助
