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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。
In the present study, spleen cells of Balb/c mice were immunized with purified hepatitis B surface antigen (HBsAg) “adr” and SP 2/0 myeloma cells were fused with PEG. The antibodies were detected by ELISA and RIA, and 9-well anti-HBs positive hybridomas were obtained. After 4 limiting dilutions, 4 monoclonal cells were established. One strain of SH_1D_9 hybridoma cells was selected for continuous passage in vitro and inoculated into the abdominal cavity of mice. The titer of anti-HBs in the tissue culture supernatant was 1:10,000 and the mouse ascites was 1:50,000. After specificity and stability identification, it was proved that anti-HBs had a sustained and good affinity for HBsAg, normal human serum had no cross reaction, and subtypes and Ig subclasses identified as anti-HBs “a” subtype Clusters, IgG-type monoclonal antibodies, hybridoma cells have an average of 104 chromosomes. This article discusses the different protocols adopted by HBsAg-immunized Balb/c mice and the experimental methods for the establishment of monoclonal antibody hybridoma cell lines.