人源单克隆抗人免疫缺陷病毒1型抗体Fab段基因的获得  

Selection of Human Monoclonal Antibody Fab Fragments to Human Immunodeficiency Virus Type 1 from Phage Antibody Library

作　　者：何红霞[1] 貌盼勇[1] 侯俊[1] 朱雷[1] 洪世雯[1] 王琰[2] 

机构地区：[1]解放军第302医院 [2]海军总医院,北京100037

出　　处：《病毒学报》2000年第4期317-321,共5页Chinese Journal of Virology

基　　金：国家自然科学基金!资助 (396 0 0 132 )

摘　　要：应用噬菌体抗体库技术有效地筛选出了多株抗HIV 1人源单克隆抗体。以逆转录聚合酶链反应 (RT PCR)从HIV 1感染者外周血淋巴细胞中扩增抗体轻重链可变区基因 ,插入载体 pCOMB3 ,建立噬菌体抗体库。分别以HIV 1gp41、gp12 0和 gp16 0为固相抗原 ,经过多轮筛选 ,从中获得了多株抗HIV 1gp41、gp12 0和 gp16 0的单克隆抗体Fab段基因。抗HIV特异性噬菌体抗体随抗体库的筛选高度富集 ,抗gp41阳性克隆率由第 2轮的 5 %增加到第6轮的 87%。间接ELISA、竞争抑制ELISA和Dotblot检测结果表明 ,获得的抗HIV 1噬菌体抗体具有较高抗原结合活性和特异性。研究结果显示 :应用该技术获得人源抗病毒基因工程单克隆抗体既省时省力 ,又可获得任意目的单抗基因 ,且易获得高亲和力的抗体。Human monoclonal antibody Fab fragments to HIV 1 have been prepared by using phage display technology. The human IgG Fab genes of heavy and light chains were amplified from an individual with HIV 1 infection. The combinatorial phage antibody library was prepared by inserting both heavy and light chain Fab genes into phagemid vector pCOMB3 and followed by the help of helper phage infection. The library was selected by panning phages, which expressed the specific human antibody Fabs on their surfaces. Six rounds of panning against coated HIV 1 gp41, gp120, and gp160, respectively were performed and showed specific enrichment of phage antibodies. The specific human Fab fragments to HIV 1 were selected and exhibited higher affinity. The specific bindings of the Fab antibodies to HIV 1 were verified by their specific reactions to HIV 1 antigen in ELISA, competitive inhibition ELISA, and Dot blot. The results provided potential promise for future use of phage display library in generation of human monoclonal antibody to HIV 1, even in prophylaxis or therapy of HIV infection.

关 键 词：噬菌体抗体库 人源单克隆抗体 人免疫缺陷病毒1 

分 类 号：R512.91[医药卫生—内科学]