人源抗狂犬病毒单克隆抗体Fab段基因的获得和表达

运用噬菌体表面呈现（phage display）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RTPCR方法，用一组人IgG Fab基因4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，该单抗为一株新的抗狂犬病毒人源基因工程抗体。

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

Human monoclonal antibody (MAb) Fab fragments to the rabies virushave 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.