小鼠抗—HBs的单克隆抗独特型抗体杂交瘤细胞系的建立及鉴定

用纯化的小鼠单克隆抗-HBs(Ab1)免疫同系BALB/c小鼠,取免疫小鼠脾细胞与Sp2/0骨髓细胞融合,获得两株(MA1、MA3)持续分泌抗-HBs的单克隆抗独特型抗体(抗-1d,Ab2)杂交瘤细胞系。对MA3做了较全面的鉴定。MA3的Ig类型为IgM,核型分析结果染色体数为106条。通过ELISA竞争抑制和中和抑制试验表明,MA3所分泌的Ab2既能被不同来源的抗-HBs所中和,又能被HBsAg所竞争,且都呈现剂量依赖关系。将纯化的MA3 IgM免疫同系小鼠,诱导出具有抗-HBs活性的抗-抗-独特型抗体(Ab3)。因此认为MA3所分泌单克隆抗体(McAb),可模似HBsAg,具有类似HBsAg的免疫原性,是一种带有HBsAg表位内影像的抗-HBs的单克隆抗-1d。

Production and Identification of Mice-mice Monoclonal Anti-idiotypic Antibody against Anti-HBs

In this paper , two hybridomas (MA1, MA3) secreting monoclonal anti-idiotypic antibodies (anti-Id) against anti-HBs were prepared by fusing SP2/0 with spleen cells from BALB/c mice immunized with syngeneic monoclonal anti-HBs to the a determinates purified by salting out and ion exchange chromatography. Competitive inhibition test and neutralization test using ELISA showed that MA3(Ab2) completely inhibited the binding of horse anti-HBs with HBsAg and could be neutralized by anti-HBs from various sources. Anti-anti-Id (Ab3) was successfully induced by peritoneally injecting purified Ab2 in BALB/c mice. Ab3 induced by MA3 specifically reacted with HBsAg. These data revealed that monoclonal anti-Id against anti-HBs produced by MA may mimic the antigenic determinant of HBsAg, possess the immunogenicity of HBsA3 and is the anti-Id bearing the internal image of HBsAg epitope.