用噬菌体展示肽库技术筛选甲肝病毒抗原模拟表位  

Hepatitis A virus mimotope mapping by phage display peptide library

作　　者：曹经瑗[1] 李建东[1] 郑惠惠[1] 毕胜利[1] 李德新[1] 

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206

出　　处：《中华实验和临床病毒学杂志》2012年第6期456-459,共4页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的用噬菌体展示肽库技术筛选甲型肝炎（甲肝）病毒抗原模拟表位，为病毒抗原决定簇定位探索可行方法。方法用纯化的抗甲肝病毒单克隆抗体，对噬菌体展示12肽库进行3轮“吸附一洗脱一扩增”筛选，随机挑取10个克隆，用酶联免疫吸附法（ELISA）对噬菌体克隆进行抗原性鉴定、竞争抑制鉴定及DNA序列测定分析，推导出展示肽氨基酸序列并与甲肝病毒（HAV）代表株结构蛋白氨基酸序列比较。结果lO个噬菌体克隆ELISA检测全为阳性，9个具有一致序列，与HAVHMl75株结构蛋白中和活性表位之一：VPl157—171区具有类似序列，另一株噬菌体克隆在HAVHMl75中未发现类似序列，结果表明这些展示肽可能是HAV抗原模拟表位。结论用噬菌体展示肽库技术筛选得到了HAV模拟表位，为开展病毒模拟表位研究打下了基础。Objective A 12 mer phage display peptide library was used to identify hepatitis A virus mimotopes of antigenic determinants, to provide the feasibility of virus epitope mapping by using this approach. Methods Using purified anti-hepatitis A virus monoclonal antibody as affinity selective molecule, phage display peptide library was biopanned and positive clones were selected by ELISA, competition assay and I）NA sequencing. Results 10 ELISA positive clones were chosen for DNA sequencing, and the displayed peptide sequences were deduced. 9 of them showed identical nucleotide sequence, and similarity in their amino acid sequence with VP1 of HAV HM175 was found, but no sequence homology was found between the other phage clone and the capsid proteins of HAV. Those peptides may behave as mimotopes of HAV. Conclusion The mimotope of HAV was selected by using phage display peptide library screening. The results provide the potential of this method to search for the mimotopes of the virus.

关 键 词：肝炎病毒 甲型 肽库 表位 

分 类 号：R3[医药卫生—基础医学]