分泌载脂蛋白AII单克隆抗体杂交瘤的建立

Establishment of hybridoma cell lines secreting anti-human apolipoprotein All monoclonal antibodies

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。

Human serum apolipoprotein AII (ApoAII) was used as the antigen to immu-nize BALB/C mice. Then the splenocytes from the immunized BALB/C mice werefused with NS--1 myeloma cells. Two hybridoma cell lines (1-C1 and 2-C2)secreting anti--human serum ApoAII monoclonal antibodies were obtained throughselective culture in HAT medium, specific screening and cloning. Immunogloblinsubclasses of the McAb secreted by two hybridoma cell lines were determined asIgG1 with immunodouble diffusion. Antibody titers in culture supernatanta of 1-C1and 2-C2 were both 1:100, while those in ascites fluids induced by the two celllines were 1:10000 and 1:5000 respectively. The McAbs showed the apecificity ofApoAII in absorption test and ELISA crossing test.The chromosome number of thetwo cell lines were 96.34±6.06 and 95.01±5.38 respectively. McAb from 1-C1 cellline culture supernatants was purified with Protein A Sepharose 4B column.