抗HBV前S_2蛋白单克隆抗体的制备及其相应抗原的免疫组化定位研究

Preparation of McAbs to Hepatitis B Virus Pre-s2 Protein and Immunohistochemical Study on its Related Antigen

本文报告用聚合人血清白蛋白(PHSA)亲和层析柱纯化具有前S_2抗原活性的HBsAg颗粒作为免疫原,通过常规融合、筛选和克隆化,获得3株分泌抗前S_2蛋白单克隆抗体杂交瘤细胞系(E8、C9和F3)。3种单克隆抗体对肝组织的免疫组化试验表明,前S_2抗原在肝炎肝硬化和肝癌周围组织中均有不同程度的表达,其染色表型,E8呈局灶胞浆型,C9为弥漫胞浆型,F3为膜型。

Three different monoclonal antibodies ( McAbs ) , designated E8, C9 and F3, were obtained by immunization of BALB/c mice with HBsAg particles with highly active pre-s2-specific antigen purified from, the plasma of chronic HBsAg carrier using the polymerized human serum albumin affinity chromatography column. The specificity of the three antibodies was determined by enzyme-linked immunosorbent assay, immun-ohistochemical test and Western immunoblotting assay, and polyclonal antibodies specific for the pre-S2 antifen were used for suppression and control tests. The results of immunohistochemical study using the three McAbs showed that pre-S2 antigen was expressed on liver tissue hepatitis B and cirrhosis and peripheral tissue of hepatocar-cinoma. The staining phenotypes of the three McAbs ( E8, C9 and F3 ) were focal cytoplasmic, diffuse cytoplasmic and cytoplasmic membrane type, respectively.