基于基因免疫的抗HBVM蛋白单克隆抗体的制备

Preparation and Characterization of Monoclonal Antibody Against HBV M Protein by Gene Immunization

目的 以基因免疫制备抗乙型肝炎病毒M蛋白的单克隆抗体。方法 构建表达M蛋白的重组质粒pcDNA3 PreS2 /S ,免疫雌性BALB/c小鼠 ,以杂交瘤技术制备单克隆抗体。 结果 目的基因片段PreS2 /S(875bp)正确插入质粒 pcDNA3,重组质粒转染细胞上清、基因注射肌肉、脾脏局部均可检测到M蛋白表达。免疫小鼠后以杂交瘤技术获得了一株分泌抗HBVM蛋白的杂交瘤细胞株 2D3。

Purpose To generate McAb against HBV M protein by gene immunization. Methods Eukaryotic expression recombinant pcDNA3-PreS2/S containing HBV M protein coding sequence was constr-ucted.Female BALB/c mice were immunized with the plasmid for preparation of specific McAb. Results The 875 bp PreS2/S gene was correctly inserted into pcDNA3.M protein could be detected in the supernat-ant of transfeced cells,local injected sites of musle and spleen.One hybridoma cell line 2D3 that could consistently produce anti-M protein monoclonal antibodies were obtained. Conclusions Monoclonal antibodies against HBV M protein could be produced by gene immunization.