人源抗EV71病毒基因工程抗体的研究  

Generation of recombinant human antibodies for EV71 virus

作　　者：孙丽娜[1] 张黎[1] 张福顺[1] 李川[1] 张全福[1] 李德新[1] 梁米芳[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,传染病预防控制国家重点实验室,北京100052

出　　处：《中华实验和临床病毒学杂志》2011年第3期161-163,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：传染病预防控制国家重点实验项目（2008SKLID103）;中国疾病预防控制中心青年科研基金（2009A104）

摘　　要：目的 研制人源抗EV71病毒基因工程抗体.方法 采集多个患儿外周血淋巴细胞,构建人源抗EV71单链（scFv）噬菌体抗体基因文库.用纯化的EV71 VPI蛋白对抗体库进行筛选,将筛出抗体的轻链和重链通过序列测定确定抗体轻重链型别后分别克隆入全抗体表达载体pAC-LCH3R后转染昆虫Sf9细胞,利用杆状病毒/昆虫细胞系统实现全抗体的分泌型表达.结果 成功地获得了1株抗EV71病毒VPI蛋白的人源单克隆抗体,利用ELISA、IFA对所获人源单克隆抗体的功能特性进行鉴定,表达成分泌型IgG全抗体在体外与A型及C4亚型EV71病毒的中和反应结果 显示为阴性.结论 从免疫库中获得了1株人源非中和单抗,为EV71病毒引起的手足口病的鉴别诊断奠定了基础.Objective To obtain recombinant human anti-EV71 antibodies from a EV71 -associated hand-foot-and-mouth disease patient-derived antibody phage library. Methods A combinatorial human scFv library to enterovirus 71 （EV71） virus was constructed using antibody genes harvested from the blood of EV71 virus patients. The library was panned and selected by using purified VP1 protein of EV71 virus with phage display. After that the specific antibody was converted to full human IgG antibody with recombinant baculovirus/insect cell system. Results One unique human scFv antibody specific for EV71 virus VP1 protein was obtained by ELISA, IFA and analysis of the antibody DNA sequence. The specific anti-VP1 human scFv antibody was converted to full human IgG antibody with recombinant baculovirus/insect cell system. The full human IgG antibody was tested in vitro for EV71 virus neutralization, resulting in no neutralizing activity with EV71 A type and EV71 C4 subtype. Conclusion The obtained human anti-EV71 antibodies without neutralizing activity laid the foundation for diagnosis of human EV71 -associated hand-foot-and-mouth disease.

关 键 词：肠道病毒属 肽库 免疫球蛋白类 轻链 免疫球蛋白类 重链 

分 类 号：R3[医药卫生—基础医学]