人源性抗HIV-1 V3环噬菌体抗体的基因克隆与序列分析

Gene cloning and sequencing of human phage antibody against HIV-1 V3 loop

以HIV1gp120V3环的合成环肽为抗原,从人的噬菌体抗体组合文库中,筛选出与HIV1V3肽具有结合活性的人源性噬菌体抗体(Fab段)。用ELISA测定其活性,竞争抑制实验证实了该噬菌体抗体的特异性。序列分析表明,重链基因为IgG1亚类,可变区属VHI亚组,与胚系基因DP88的同源性最高,D区为D33,J区为JH5;轻链为κ亚型,可变区属VLIV亚群,D区为DPK22,J链为JK4胚系。

Aphageantibody (Fab)againstHIV 1gp1 2 0V3loopwasclonedbyscreeningahumanimmunoglobulincom binatoriallibrary.TheV3 bindingactivityandspecificityoftheantibodywereassayedbyELISAandcompetitiveinhibi tionELISA .ComparisonoftheFdgeneandlightchaingenewiththatinKABATdatabaseshowedthattheFdbelonged toIgG1subclass.ThevariableregionofheavychainwasVHIsubgroup,derivedfromrearrangedgerm linegeneof DP88.Whilethelightchainbelongedtoκsubtype.ThevariableregionwasVLIVsubgroup,derivedfromthegerm line geneDPK2 2 .