抗TNF-α单克隆抗体对噬菌体随机12肽库的筛选  

Screening of Phage Displayed Random Dodecapeptide Library for Mimicking Epitopes by Anti-TNF-α Monoclonal Antibody

作　　者：龙剑[1] 谢红付[1] 施为[1] 

机构地区：[1]中南大学湘雅医院皮肤科,湖南长沙410008

出　　处：《医学临床研究》2003年第4期248-251,共4页Journal of Clinical Research

基　　金：湖南省自然科学基金资助项目 (项目编号 :0 1JJY2 0 15 )

摘　　要：目的试从噬菌体随机多肽库中筛选出能与肿瘤坏死因子 α(TNF α)特异性结合的短肽分子。方法采用抗TNF α单克隆抗体作为配基 ,免疫亲和筛选以融合蛋白形式在丝状噬菌体M13 外壳蛋白Ⅲ表达的随机 12肽文库 ,按“吸附 -洗脱 -扩增”的淘洗过程 ,经过 3轮免疫筛选后 ,随机挑取阳性噬菌体克隆用ELISA及斑点ELISA检测其特异性。结果经过 3轮免疫筛选 ,特异性结合的阳性噬菌体克隆富集增加近10 0倍 ,第 3轮筛选后随机挑选 7个阳性噬菌体克隆经ELISA和斑点ELISA检测 ,有 5个克隆能与抗TNF α特异性结合。结论噬菌体随机肽库技术可以应用于分析、鉴定抗TNF α的表位 ,继而为获得亚单位表位水平的诊断试剂和抗TNFObjectivesTo screen out the short peptide molecule capable of combining with tumor necrosis factor α(TNF α) from phage displayed random dodecapeptide library.MethodsUsing TNF α as ligand, a phage displayed random peptide library consisting of 12 amino acid residues was immunoscreened . This library expressed as a fusion protein inside the coat protein Ⅲ of filamentous phage M13 .According to the process of “absorption elution amplification”, the positive clones were obtained by three rounds of biopanning. The specificity of each clone binding to anti TNF α was detected by ELISA and dot ELISA.ResultsThe eluted phages were enriched nearly 100 fold through three rounds of biopanning, seven positive phage clones from the third round biopanning were randomly selected and 5 clones of them could specifically bind to the anti TNF α.ConclusionThe phage display technique can be applied to study the anti TNF α antigenic peptides, and these epitopes can provide the potential for developing immunodiagnostic reagents on a level of subunit expression and preparing of anti TNF α short peptide vaccines. [

关 键 词：抗TNF-Α 单克隆抗体 噬菌体 随机12肽库 筛选 肿瘤坏死因子-Α 

分 类 号：R392－33[医药卫生—免疫学] R373.9[医药卫生—基础医学]