抗人免疫缺陷病毒1型gp120人源单链抗体的表达  

Expression of human ScFv against human immunodeficiency virus type 1 gp120

作　　者：何红霞[1,2] 貌盼勇[1,2] 朱雷[1,2] 侯俊 洪世雯[1,2] 阎锡蕴 

机构地区：[1]解放军302医院病毒研究室 [2]中科院微生物所

出　　处：《中华实验和临床病毒学杂志》1998年第3期233-236,共4页Chinese Journal of Experimental and Clinical Virology

基　　金：国家自然科学基金

摘　　要：目的研究人源抗人免疫缺陷病毒１型（ＨＩＶ－１）ｇｐ１２０单链抗体（ＳｃＦｖ）。方法以人工合成的ＨＩＶ－ｌｇｐ１２０Ｖ３环多肽为抗原，利用噬菌体抗体库技术，筛选含有抗－ｇｐ１２０ＳｃＦｖ基因的噬菌体，提取质粒，转化大肠杆菌ＨＢ２１５１，表达可溶性ＳｃＦｖ。结果经ＳＤＳ－ＰＡＧＥ和Ｗｅｓｔｅｒｎｂｌｏｔ分析，表达产物分子量为２８ｋＤ左右，且具有ｃ－ｍｙｃ活性；ＥＬＩＳＡ和Ｄｏｔｂｌｏｔ结果表明，可溶性ＳｃＦｖ具有较好的抗原结合活性和较强的特异性；竞争性ＥＬＩＳＡ实验结果进一步证明表达产物的特异抗－ｇｐ１２０活性。结论该技术便捷有效，可大量获得人源抗ＨＩＶ抗体片段。In order to generate human antibody against HIV-1, using synthetic peptide from HIV-1 gp120 v3 loop as antigen, we screened the recombinant phage antibodies by immunoaffinity and selected positive clones carrying anti-gp120 ScFv gene. The phage DNA was extracted and transformed to E.coli HB2151 that allowed to express soluble ScFv proteins by inducing of IPTG. The results of SDS-PAGE and Western blot showed that the molecular weight of the expressed product is about 28KD and exhibited anti-c-myc activity. ELISA and Dot blot analysis displayed that the soluble ScFv specifically bound to gp120 and had a higher specificity and affinity. Competitive ELISA further demonstrated that the product was specific to gp120.

关 键 词：HIV-1 外膜糖蛋白 GP120 人源单链抗体 表达 

分 类 号：R512.910.5[医药卫生—内科学] R392－33[医药卫生—临床医学]