利用噬菌体随机肽库技术分析丙肝患者血清中抗HCV抗体的表位  

Epitope mapping of anti-HCV antibodies from patients serum

作　　者：柯屾[1] 戚中田[2] 温新宇[2] 施明[3] 胡美茹[3] 沈倍奋[3] 

机构地区：[1]解放军总医院基础医学研究所,北京100853 [2]第二军医大学微生物学教研室,上海200433 [3]军事医学科学院基础医学研究所,北京100850

出　　处：《军医进修学院学报》2002年第4期283-285,共3页Academic Journal of Pla Postgraduate Medical School

基　　金：国家自然科学基金资助重点项目 ( 39830 330 ) ;国家"86 3"计划资助项目 ( 10 2 0 9 0 3 0 4)

摘　　要：目的 :分析患者血清中抗HCV抗体的抗原表位。方法 :用ProteinA亲和层析柱从患者血清中纯化、制备抗HCV多克隆抗体 ;以此对噬菌体表面展示的随机 6肽库进行亲和筛选。结果 :经具有良好富集效果的三轮筛选后 ,从第三轮挑选出的 12个克隆进行结合试验 ,发现 8个克隆只与HCV抗体有较强的结合力而不与正常人血清反应 ;测序表明阳性克隆的外源肽具有一定的相似性。用阳性噬菌体克隆检测 2 0例患者血清 ,有程度不等的阳性反应。结论 :用多克隆抗体从噬菌体随机肽库中筛选得到了有一定功能的模拟表位。Objective:epitope analysis of anti-HCV antibodies isolated from patients serum. Methods: Anti-HCV Abs were purified from patients sera using an affinity chromatography column which constructed with sepharose 4B and a recoment HCV multi-epitope antigen. These Abs were used for biopanning of a phage-displayed random peptide library of 6 amino acid residues. Results:After 3 rounds of effective screening, twelve clones were selected for binding test with Abs from patient′s and normal sera. Eight clones was proved to specifically react with the sera from patient. The positive phage clones could react with different patients sera and not react with normal sera. Conclusion:It is possibility that the sequences in the short peptide are the mimic of HCV epitopes that can be recognized by anti-HCV Abs.

关 键 词：噬菌体随机肽库 丙肝 血清 抗HCV抗体 表位 

分 类 号：R512.63[医药卫生—内科学]