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以人血清载脂蛋白AⅡ为抗原,免疫BALB/C小鼠,将脾细胞与NS-1骨髓瘤细胞融合,经HAT选择性培养,特异性筛选及克隆化,获得1-C1、2-C2两株分泌抗人血清载脂蛋白AⅡ单克隆抗体杂交瘤细胞株。免疫双扩散确定它们分泌抗体的亚型均为IgG1;吸收试验及ELISA交叉试验表明单克隆抗体的特异性;ELISA测定其培养上清中抗体效价都为1:100,诱生腹水效价分别为1:10000和1:5000;两株杂交瘤细胞染色体均数分别为96.34±6.06、95.01±5.38;采用Protein A Sepharose 4B层析柱纯化了1-C1杂交瘤细胞培养上清中的单克隆抗体。
BALB / C mice were immunized with human serum apolipoprotein AⅡ as antigen, spleen cells were fused with NS-1 myeloma cells, selectively cultured and selectively cloned by HAT to obtain 1-C1,2-C2 Two monoclonal antibodies secreting anti-human serum apolipoprotein A II hybridoma cell lines. The double immunodiffusion assay showed that the subtype of antibody secreted by them was IgG1. Absorption test and ELISA cross-test showed the specificity of monoclonal antibody. The titer of antibody in culture supernatant was 1: 100 by ELISA, respectively, 1: 10000 and 1: 5000 respectively. The average number of chromosomes in two hybridoma cells was 96.34 ± 6.06 and 95.01 ± 5.38, respectively. The monoclonal antibodies in 1-C1 hybridoma cell culture supernatant were purified by Protein A Sepharose 4B column antibody.