Takebayashi T
Hokkaido Igaku Zasshi. 1982 Nov;57(6):667-78.
Somatic cell hybrids which produced antibodies against HBsAg were established by fusing the splenocytes of BALB/c mice immunized with purified HBsAg and the myeloma cell line P3X63Ag8 using polyethylene glycol 1,000. Mouse ascites were obtained by inoculating 13 hybridoma clones, and 8 of them produced high PHA titer antibodies of 10(2) to 10(6) against HBsAg. These antibodies in ascites were analysed by way of Sephadex G-200 gel filtration, micro-Ouchterlony method and 2ME, DTT treatment. According to these analyses, one of these antibodies has belonged to IgM and the other 7 to IgG. By RPHA titration using tannic acid-treated SRBC coated with these antibodies, it was suggested that 2 clonal antibodies were against the common antigenic components of HBs subtypes and the other 5 against the component r. According to the results of RPHA test and PHA suppression test using these clonal antibodies and 116 samples of HBsAg positive sera in Hokkaido, one subtype of HBsAg was further classified into several groups. These antibodies will be valuable for many virological and serological investigation, for instance, for the detailed antigenic analyses of HBsAg and for searching a source of HBs infection containing feto-maternal infection.
用聚乙二醇1000将经纯化的乙肝表面抗原(HBsAg)免疫的BALB/c小鼠脾细胞与骨髓瘤细胞系P3X63Ag8融合,建立了产生抗HBsAg抗体的体细胞杂种。通过接种13个杂交瘤克隆获得小鼠腹水,其中8个产生了效价为10(2)至10(6)的高PHA效价抗HBsAg抗体。用Sephadex G - 200凝胶过滤法、微量双向免疫扩散法及2 - 巯基乙醇(2ME)、二硫苏糖醇(DTT)处理法对腹水中的这些抗体进行分析。根据这些分析,其中一种抗体属于IgM,另外7种属于IgG。用包被有这些抗体的经鞣酸处理的绵羊红细胞进行反向被动血凝试验(RPHA)滴定,结果表明2种克隆抗体针对HBs亚型的共同抗原成分,另外5种针对成分r。根据使用这些克隆抗体及北海道116份HBsAg阳性血清进行的RPHA试验和PHA抑制试验结果,将一种HBsAg亚型进一步分为若干组。这些抗体对于许多病毒学和血清学研究将是有价值的,例如,用于HBsAg的详细抗原分析以及寻找包括母婴感染在内的HBs感染源。