抗戊型肝炎病毒重组蛋白单克隆抗体的制备和初步应用

Preparation and Application of Monoclonal Antibody Against Recombinant Hepatitis E Virus Protein

目的:制备抗-戊型肝炎病毒的单克隆抗体,并将其用于分析戊型肝炎病毒不同毒株结构蛋白的抗原表位。方法:采用来自墨西哥株(Mexicanstrain)的戊型肝炎病毒重组蛋白(p166Mex)免疫Balb/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞进行融合,经酶联免疫吸附试验(ELISA)法筛选阳性克隆,并将获得的单克隆抗体与戊型肝炎病毒缅甸株(Burmastrain)和美国株(USAstrain)的重组蛋白(p166Bur、p166US)进行交叉反应测定。结果:最终获得4株能稳定分泌抗-p166Mex的杂交瘤细胞株,即D8G10、E5E12、D4A3、B7E6。其中D8G10,E5E12和B7E6细胞株的培养上清液,还能分别与p166Bur和p166US重组蛋白发生阳性反应。结论:利用已获得的抗-p166Mex单克隆抗体,初步确定3种不同的戊型肝炎病毒重组蛋白(p166Bur、p166US、p166Mex)含有一种共同的抗原表位。

Objective:To prepare McAbs against HEV and study the antigenic epitopes of different HEV strains.Methods:The recombinant protein(p166Mex)from Mexican prototype virus was immunized to Balb/c mouse,and the spleen cells of the immunized mouse were fused with SP2/0 myeloma cells.The positive clones which produced McAbs against p166Mex were screened by ELISA.The cross reactions of the McAbs were tested with the recombinant proteins(p166Bur、p166US、p166Mex)from different HEV strains.Results:Four hybridoma cell lines (D8G10、E5E12、D4A3、B7E6) secreting specific McAbs against p166Mex were chosen.All McAbs,but one,specific for p166Mex were strongly reactive with p166Bur and p166US.Conclusion:With McAbs directed against p166Mex,we suggested that there is a common antigenic epitope among the three different recombinant proteins of HEV.