用噬菌体随机肽库筛选SARS冠状病毒S蛋白单克隆抗体2C5的模拟表位  

Identification of Mimotope Peptides Which Bind to the SARS-CoV Spike Protein Specific Monoclonal Antibody 2C5 with Phage-Displayed Peptides Library

作　　者：华荣虹[1] 吴东来[1] 童光志[1] 王云峰[1] 田志军[1] 周艳君[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,哈尔滨150001

出　　处：《生物工程学报》2006年第5期701-706,共6页Chinese Journal of Biotechnology

基　　金：黑龙江省博士后基金资助(No.HPD2003045)。~~

摘　　要：SARS-CoVS蛋白特异的单克隆抗体2C5具有病毒中和作用。以单克隆抗体2C5为筛选靶分子,筛选噬菌体展示随机7肽库。经三轮淘洗后随机挑选20个噬菌体克隆进行ELISA分析和序列测定。在10个ELISAOD值大于0.2的阳性噬菌体克隆中,有8个噬菌体克隆展示有共同的7肽序列TPEQQFT。展示有该序列的噬菌体克隆能竞争抑制SARS-CoVS蛋白抗原与单抗2C5的结合。结果表明TPEQQFT为单克隆抗体2C5的模拟表位。该结果可对进一步研究S蛋白结构与功能和设计SARS疫苗有一定的参考意义。To identify the epitope of SARS-CoV spike protein specific neutralizing monoclonal antibody （MAb） 2C5. The antibody was used as target and three rounds of bio-panning were conducted with phage-display peptide library. After the third panning, 20 phage-plague clones were randomly picked and analyzed for the binding ability with the MAb 2C5 by ELISA. The display sequence analysis demonstrated that among the twenty phage clones, eight clones displayed the same seven-peptide TPEQQFT. All these eight phage-clones showed strongest binding activity with 2C5 in phage ELISA analysis. Furthermore, phages displaying peptide TPEQQFT could specifically inhibit the binding of MAb 2C5 with SARS-CoV spike protein. The results demonstrated that TPEQQFT is a mimic epitope peptide containing neutralizing MAb 2C5. This study may provide information for further structural and functional analysis of spike protein and development vaccine for severe acute respiratory syndrome.

关 键 词：SARS冠状病毒(SARS-CoV) S蛋白 单克隆抗体 噬菌体随机肽库 模拟表位 

分 类 号：Q789[生物学—分子生物学]