人源单克隆抗人免疫缺陷病毒1型抗体Fab段基因的获得

应用噬苏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－１ｇｐ１２０和ｇｐ１６０为固相抗原,经过多轮筛选,从中获得了多株抗ＨＩＶ－１ｇｐ４１、ｇｐ１２０和ｇｐ１６０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

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

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.