人源抗HAV噬菌体抗体的筛选及基因序列分析

Selection of human phage antibodies against HAV and DNA sequence analysis

目的获得人源性甲型肝炎病毒（ＨＡＶ）抗体。方法应用抗体捕获的ＨＡＶ，从本实验室构建的噬菌体抗体组合文库中筛选与ＨＡＶ病毒有结台活性的人源噬菌体抗体（Ｆａｂ段），用夹心ＥＬＩＳＡ和竞争抑制实验测定其活性及特异性。结果筛选出与ＨＡＶ病毒有结台活性和特异性的人源抗体。序列分析表明重链基因为ＩｇＧＩ亚类，可变区属ＶＨＩ亚群，是ＤＰ８８、Ｄ３—３和ＪＨＳ胚系基因片段的重排产物；轻链基因属ＶＸｌｌｌ亚群，是ＤＰＸ２２和Ｊｋ４胚系基因的重排产物。结论应用抗体捕获抗原的方法，能从该抗体库筛选出ＨＡＶ抗体，也说明该抗体库的构建是成功的。

Objective To obtain the anti-HAV antibodies. Methods The antibody--capturing antigenstrategy was used in panning antibody library with impure antigen. The HAV--binding activity and speCificity of ptiagr antibodies were assayed by sandwich ELISA and competitive inhibition ELISA. ResultsPhagr antibodies(Fab) against HAV were screened from a human immunoglobulin combinatorial library.Fd and light chain gmes were wnuenced. Comparison of the Fd bine and light chain gene with KABATdatabase showed that the heavy chain belonged tO IgGI subclass, and its variable region was derived fromrearranged farm--line ace of DP88. D3--3 and JHS; while the light chain was a member of VkIII family,the farm-line gene was DPk22 and Jot. Conclusion Phals antibodies against HAV were selected from theantibody library by using the antibody--capturing antigen strategy. The result also suggested that theantibody library was condruct6d successfuny