抗SARS-CoV人源Fab抗体的表达及鉴定  

Expression and Identification of Human Fab Antibody Against Spike Protein of SARS-CoV

作　　者：李郁[1] 陈江浩[2] 包国强[3] 张思河[1] 宋斐[1] 谢丽[1] 杨红[2] 

机构地区：[1]第四军医大学细胞工程研究中心,西安710032 [2]西京医院,西安710032 [3]唐都医院,西安710032

出　　处：《科学技术与工程》2006年第11期1470-1473,共4页Science Technology and Engineering

基　　金："973"项目(2003CB514127)资助

摘　　要：为进一步研究抗SARS-CoV刺突蛋白S(Spike)的中和抗体2g7,对其进行了原核表达,将2g7转入琥珀突变的非抑制性菌株(SupE-)的大肠杆菌Top10F'中进行可溶性表达。用HPLC纯化后,SDA-PAGE显示2g7的轻重链均存在,ELISA,Westernblot检测显示2g7能与SARS-CoV的S1蛋白结合。BIAcore测定其与抗原的亲和力为2.67×10-8mol。竞争ELISA显示该抗体能与SARS-CoV刺突蛋白S(Spike)结合并阻断该蛋白与其受体ACE2的结合。实验结果表明:利用免疫噬菌体抗体库能在小库容的情况下筛选获得高亲和性的人源抗体,并可以在大肠杆菌中获得高效表达,这一实验结果为有效性地防治病毒性疾病提供了思路。To further investigate function of human Fab 2g7 against spike protein of SARS-CoV, expressed in E.coli. Genes encoding 2g7 were transferred to amber suppressor free strain（SupE-） Top10F＇to be translated to soluble protein. Fab 2g7 was purified by High Performance Liquid Chromatography （HPLC）. Result of SDSPAGE showed that expressed product contained light chain（LC） and heavy chain（HC）. It can bind to S1 protein of SARS-CoV assayed by ELSIA and Western blot. Compete ELISA showed that 2g7 can block spike protein of SARS-CoV binding to its receptor ACE2. Equilibrium dissociation constant （Kd） of 2g7 was 2.67×10^-8mol measured by BIAcore. Results above indicate that human antibody of high affinity can be obtained by screening immune phage antibody library with small capacity. They also suggest new approaches to prevent and treat virus disease.

关 键 词：严重急性呼吸综合征(SARS) 刺突蛋白 噬菌体抗体 FAB 

分 类 号：R511[医药卫生—内科学]