人源抗狂犬病毒单克隆抗体Fab段基因的获得和表达  

Generation of Human Monoclonal Antibody Fab Fragment to Rabies Virus by Phage Display Technology

作　　者：张世珍[1] 梁米芳[1] 董关木[2] 郑海法[2] 侯云德[1] 

机构地区：[1]中国预防医学科学院病毒学研究所病毒基因工程国家重点实验室,北京100052 [2]中国药品生物制品检定所,北京100050

出　　处：《病毒学报》2001年第1期11-16,共6页Chinese Journal of Virology

摘　　要：运用噬菌体表面呈现 (phagedisplay)技术获得了人源抗狂犬病毒糖蛋白基因工程单克隆抗体Fab段基因及其表达。从狂犬病毒PM株Vero细胞疫苗免疫的人抗凝血中分离获得外周淋巴细胞 ,提取细胞总RNA ,通过RT -PCR方法 ,用一组人IgGFab基因特异性引物 ,从合成的cDNA中扩增了一组轻链和重链Fab段基因 ,将轻链和重链先后克隆入噬菌体载体 pComb3,成功地建立了抗狂犬病毒噬菌体抗体库 ,并用纯化的狂犬病毒颗粒和几株抗狂犬病毒鼠单克隆抗体 (单抗 )捕捉狂犬病毒抗原的方法 ,对此抗体库进行富积筛选表达 ,成功地获得了抗狂犬病毒的人源单抗Fab段基因及其在大肠杆菌中的有效表达。对其中一株单抗G10进行了较为系统的分析 ,发现它与一株鼠源中和性狂犬病毒糖蛋白特异性单抗存在竞争 ,证实该单抗能识别狂犬病毒糖蛋白。其序列资料分析表明 。Human monoclonal antibody (MAb) Fab fragments to the rabies virus have been developed by using phage display technology for the first time. The genes of antibodies to rabies virus were amplified from the donors vaccinated by the vaccine of PM strain produced in Vero cell line. The combinatorial phage antibody libraries were constructed by inverting the genes of light chain and Fd chain into pComb3 vector. The libraries were panned and selected by using purified rabies virus antigen of aG strain with phage displaying. The MAbs′ capture method was also used in the screening process. Three recombinant human Fab antibodies to rabies virus were obtained finally in a short period. Among the three antibodies, Fab antibody G10 was identified against rabies virus glycoprotein by competition with a mouse MAb to GP. The variable region genes of the heavy and light chains of the Fab antibodies to rabies virus were sequenced and identified as human Fab gene.

关 键 词：狂犬病毒 噬菌体 表面呈现 人源基因工程抗体 FAB段 单克隆抗体 

分 类 号：R373.9[医药卫生—病原生物学] S852.659[医药卫生—基础医学]