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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。
Multiple phage antibody library technology was used to screen out multiple anti-HIV human monoclonal antibodies. Antibody light chain heavy chain variable region gene was amplified by reverse transcriptase polymerase chain reaction (RT PCR) from HIV-1 peripheral blood lymphocytes and inserted into vector pCOMB3 to construct a phage antibody library. The monoclonal antibodies against HIV 1 gp41, gp12 0 and gp16 0 were obtained by multiple rounds of screening using HIV 1 gp41, gp12 0 and gp16 0 as solid phase antigens, respectively. Anti-HIV-specific phage antibodies were highly enriched with antibody library screening, and the rate of anti-gp41-positive clones increased from 5% in round 2 to 87% in round 6. Indirect ELISA, competitive inhibition ELISA and Dotblot assay showed that the obtained anti-HIV 1 phage antibody had higher antigen binding activity and specificity. The results show that: The application of this technology to obtain human antiviral genetically engineered monoclonal antibodies not only saves time and effort, but also to obtain any monoclonal antibody of interest, and easy access to high-affinity antibodies.