乙型肝炎表面抗原亚型决定簇“a”单克隆抗体杂交瘤细胞株的建立

本研究中用纯化乙型肝炎表面抗原(HBsAg)“adr”免疫Balb/c小鼠的脾细胞与SP 2/0骨髓瘤细胞在PEG作用下融合。以ELISA和RIA法检测抗体,获得9孔抗-HBs阳性的杂交瘤,经过5次有限稀释,建立4株单克隆细胞。选其中一株SH_1D_9杂交瘤细胞进行体外连续传代,接种小鼠腹腔,抗-HBs滴度在组织培养上清液中为1∶10,000,小鼠腹水为1∶50,000。经特异性、稳定性鉴定,证明其抗-HBs对HBsAg具有持续的和良好的亲和性,正常人血清无交叉反应,经亚型及Ig亚类鉴定为抗-HBs“a”亚型决定簇,IgG型单克隆抗体,杂交瘤细胞染色体平均为104个。本文并对HBsAg免疫Balb/c小鼠采取的不同方案及建立单克隆抗体杂交瘤细胞株的实验方法加以讨论。

The establishment of a monoclonal antibody hybridoma cell line of HBsAg subtype determinant "a"

In this study, the spleen cells of BALB/c mice which were immunised with purified HBsAg/adr were fused with myeloma cells SP2/0 under the influence of PEG. Antibodies were tested with ELISA and RIA, and 9 wells of anti-HBs-positive hybridoma were obtained. After limited dilution for 5 times 4 monoclnal cell lines were established, one of which, SH1 D9 hybridoma cell, was chosen for in vitro serial culture and intraperitoneal inoculation of mice. The titre of anti-HBs in the supernatant of tissue culture was 1: 10,000 and that in the ascitic fluid of mice was 1: 50,000. Its specificity and stability were assessed, and these tests demonstrated that the anti-HBs had long-lasting and satisfactory affinity to HBsAg and gave no false reaction with normal human serum. It was identified to be anti"a" , IgG monoclonal antibody. The chromosomes in the hybridoma cells were found to be 104 in number in average. In this paper we discussed and commented the different programmes of immunization of BALB/c mice with HBsAg and the experimental methods of establishing a hybridoma cell line of monoclonal antibody.