禽流感病毒M1蛋白型特异性表位分析

目的对流感病毒基质蛋白1(M1)型特异性表位进行定位研究。方法采用针对禽流感病毒M1蛋白的型特异性单克隆抗体,淘选M13噬菌体展示的12肽随机肽库,进行M1蛋白表(拟)位分析。采用ELISA、竞争性ELISA、免疫印迹分析不同噬菌体拟位与单克隆抗体的免疫反应性。结果筛选获得共有序列NxPHLR,定位于M1蛋白222-224位(HPN)、229-230位(LR)氨基酸区域。含有NxPHLR基序的噬菌体拟位能与单克隆抗体发生特异性结合,且此结合能被天然病毒抗原抑制或阻断,表明拟位多肽真实模拟了天然病毒蛋白与单克隆抗体结合的抗原决定簇或表位。结论M1蛋白222-230位氨基酸(HPNSSARLR)序列构成了流感病毒型特异性表位。

Mapping of the type-specific epitope in matrix protein 1(M1)of avian influenza virus

To analyze the type-specific epitope mapping or localization of matrix protein 1(M1)in avian influenza virus,the type-specific monoclonal antibodies(Mab) against M1 protein were used to screen a 12-mer random peptide phage display library for epitope mapping,and the immuno-reactivities of different phage mimotopes were analyzed by using ELISA,competitive ELISA and Western blotting. It was demonstrated that the consensus sequence NxPHLR screened was found to be located at the 222-224 position(HPN)and 229-230 position(LR)of M1 protein. The minotopes with NxPHLR motif could bind with Mab against M1 protein and this binding could be blocked or inhibited by the natural viral antigen,indicating that the mimotopes truly mimicked the Mab-binding determinants or epitopes on the natural viral antigen. It is concluded that the amino acid sequence at 222-230 position(HPNSSARLR)of M1 protein constitutes the type-specific epitope of avian influenza virus.