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以重组人红细胞生成素 (r Hu EPO)免疫 BALB/c小鼠 ,通过淋巴细胞杂交瘤技术将BALB/c小鼠脾细胞和 Sp2 /0骨髓瘤细胞融合 ,经 ELISA法筛选及有限稀释法克隆化 ,制备了 2株能稳定分泌抗 r Hu EPO单克隆抗体 (Mc Ab)的小鼠杂交瘤细胞系 G50和 A94。用鼠阊我隆抗体分型试剂盒鉴定 ,其分泌的 Mc Ab亚类均是 Ig G1 。经 ELISA鉴定 ,两株 Mc Ab只与 r Hu EPO特异性结合 ,与 IL- 1、GM- CSF、IFN- α和 IFN- r等细胞因子均无效叉反应。 Western- blot免疫印迹分析表明 ,两株 Mc Ab对 r Hu EPO有较高的亲和力
BALB / c mice were immunized with recombinant human erythropoietin (rhu EPO), BALB / c mouse splenocytes and Sp2 / 0 myeloma cells were fused by lymphocyte hybridoma technique, cloned by ELISA and diluted by limiting dilution Two murine hybridoma cell lines, G50 and A94, that stably secreted anti-Hu Hu monoclonal antibodies (Mc Ab) were prepared. It was identified by the Mycoplasma pneumoniae typing kit and its secreted McAb subclasses were both IgG1. The results of ELISA showed that the two McAbs specifically bound to r Hu EPO and did not cross-react with cytokines such as IL-1, GM-CSF, IFN-α and IFN-γ. Western-blot Western blot analysis showed that the two Mc Abs had higher affinity for rHuEPO