从人源大容量天然抗体库中筛选到抗禽流感病毒H5N1单链抗体  

Human antibody against avian influenza A (H5N1) virus from human na■ve antibody library

作　　者：孙丽娜[1] 于礼[1] 张黎[1] 李川[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,北京102206

出　　处：《中国人兽共患病学报》2011年第2期86-90,共5页Chinese Journal of Zoonoses

基　　金：青年科学基金项目(No.30801056);传染病预防控制国家重点实验项目(No.2008SKLID103);中国疾病预防控制中心青年科研基金(No.2009A104)联合资助

摘　　要：目的从人源大容量天然抗体库中筛选到抗禽流感病毒H5N1单链抗体。方法运用噬菌体表面呈现技术,用纯化的H5N1禽流感血凝素蛋白HA(A/Anhui/1/2005)对人源大容量天然抗体库进行富集筛选,对获得的阳性克隆进行序列分析,并将其克隆入携带人源Fc段的表达载体,实现scFv-Fc融合抗体的分泌型表达。用ELISA、IFA对所获人源单抗的功能特性进行鉴定。结果经过3轮富集筛选,共获得了82株特异性结合H5N1禽流感病毒HA的人源scFv单链抗体,分别属于5个不同的抗体序列,ELISA、IFA表明从抗体库筛选获得的5株人源单抗与禽流感病毒H5N1及其HA具有较好的特异性,而与甲1型和甲3型人流感病毒无交叉反应。结论从人源大容量天然抗体库中利用高通量的筛选策略在短时间内获得抗禽流感病毒H5N1人源单抗,对于禽流感的紧急预防和治疗具有重要意义。To obtain human antibody against avian influenza A（H5N1） virus from human na■ve antibody library.In our study,using phage display technology,antibodies against purified HA protein（A/Anhui/1/2005） were screening from a human na■ve antibody library.Antibody genes were identified by sequence analysis and then the scFv antibody genes were cloned into vector pCMV-hIgG1-Fc-XP and expressed in 293T cells,and secreted scFv-Fc antibodies were further identified by ELISA and IFA assay.After three rounds of panning,82 human scFv clones which demonstrated specific reactivity with HA（A/Anhui/1/2005） were selected by phage ELISA.Five different antibody genes were identified by sequence analysis of 82 clones.And five human antibodies were found to bind HA（A/Anhui/1/2005） with high stability and specificity without cross-reaction with the other substypes of influenza A viruses（H1N1,H3N2） by ELISA and IFA.Generation of recombinant human antibodies for targeting highly pathogenic avian influenza A（H5N1） virus from human na■ve antibody library in short time bring new prospect for the prophylaxis or treatment of H5N1 virus infection.

关 键 词：禽流感病毒 噬菌体表面呈现 人源大容量天然抗体库 单链抗体 

分 类 号：R373[医药卫生—病原生物学]