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应用两种在pTrcHis上表达的融合蛋白rp15 0C和rp5 2C免疫BALB/c小鼠后 ,获得 2株分泌抗pTrcHis载体蛋白单克隆抗体的杂交瘤细胞 1C4和 3C10。其中 1C4为IgG3 ,腹水ELISA效价为 1∶2× 10 6;3C10为IgM ,腹水ELISA效价为 1∶6 0 0。实验表明 ,1C4分泌的抗体用于pTrcHis表达蛋白的检测具有很好的特异性 ,对工程菌体蛋白无非特异性反应。辣根过氧化物酶HRP标记 1C4后 ,用于组装HCMVIgM检测试剂盒 ,检测HCMVIgM阳性血清 ,得到了满意的结果
After immunizing BALB / c mice with two fusion proteins rp15 0C and rp5 2C expressed on pTrcHis, two hybridoma cells 1C4 and 3C10 secreting anti-pTrcHis carrier protein monoclonal antibodies were obtained. Among them, 1C4 was IgG3, ascites ELISA titer was 1 × 2 × 10 6, 3C10 was IgM, and ascites ELISA titer was 1: 600. Experiments show that, 1C4 secretion of antibodies for the detection of pTrcHis expression of protein has a very good specificity, non-specific non-specific engineering bacteria protein reaction. HRP-labeled horseradish peroxidase 1C4, for the assembly of HCMVIgM detection kit, detection of HCMVIgM-positive serum, and obtained satisfactory results