抗HIV-1 gp160独特型阳性抗体的序列分析及表达

为了确定从噬菌体抗体文库中筛选出的抗体的属性和方便目的基因的表达及其产物的纯化，对两株具有“１Ｆ７”独特型的抗ＨＩＶ－１ｇｐ１６０抗体基因进行了序列分析并构建了可溶性表达载体．发现３Ｂ株含有完整的Ｆａｂ段，１Ｄ株只有重链Ｆｄ段．序列测定表明两株克隆的Ｆｄ段基因完全相同，其可变区ＶＨ属于ＶＨⅠ亚群，而３Ｂ株的“轻链”序列与已知的人的κ和λ轻链无同源性．用从另外的Ｆａｂ抗体文库中筛选出来的３株抗乙肝表面抗原抗体的轻链与３Ｂ的重链重组，并选择一个ＨＩＶ－１ｇｐ１６０特异性较好的重组抗体株，命名为３Ｂｓ．构建了１Ｄ株与３Ｂｓ株的可溶性表达载体，免疫印迹实验证实了具有“１Ｆ７”独特型的抗ｇｐ１６０独特型阳性抗体的表达．

Analysis and Expression of Anti HIV 1 Idiotypic Antibody

In order to classify the phagemid antibodies and to purify them simply,two human anti HIV 1 idiotypic antibodies were analyzed by restriction enzymatic mapping and DNA sequencing,and soluble expression vector was constructed.Phagemid 1D was found only containing a Fd segment.Both of the Fd segments were the same.The VH gene was derived from VHⅠ subgroup,JH from HUMJH6.The D segment contained D D fusion.The “light chain” of 3B did not belong to any of κ or λ chain families.After reconstitution the Fd with other λ light chains of anti HBsAg Fab,a new clone named 3Bs was sorted out which exhibited tighter binding ability of gp160 than to HBsAg.Soluble antibodies of 3Bs and 1D were expressed in the bacterial culture.The specificity of the antibodies was proved by immunoblot assay.