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用多肽固相合成仪合成人癌基因产物p185HER-2(即p185)胞内区的肽段,以之与沙门氏裸菌偶联制成免疫原,经脾内、腹腔、静脉途径免疫Balb/c小鼠,获得具有高抗体活性的脾细胞,后者与SP2/0融合,筛选出5株杂交瘤细胞系并对其分泌的单克隆抗体(McAb)特性进行了初步分析,ELISA检测腹水效价为10-4~10-6,用多肽和沙门氏裸菌分别进行中和试验,多肽抑制率在50%以上,而沙门氏裸菌则不能中和抗体反应。测定其中两株McAb亲和常数(Ka)分别为2.5×107M-1,5.0×108M-1。表位分析结果表明这两株McAb针对同一抗原表位
Peptide solid-phase synthesizer was used to synthesize the intracellular domain of human oncogene product p185HER-2 (p185), which was conjugated with Salmonella spp. To make immunogen. Balb / c mice were immunized intraperitoneally, intraperitoneally and intravenously. c mice. The spleen cells with high antibody activity were obtained, which were fused with SP2 / 0. Five hybridoma cell lines were screened and the characteristics of monoclonal antibodies secreted by them were analyzed. The price was 10-4 ~ 10-6. The neutralization test was carried out with the polypeptides and Salmonella spp. Respectively. The inhibition rate of the polypeptide was above 50%, but the Salmonella spp. Could not neutralize the antibody reaction. The McAb of two McAbs were determined to be 2.5 × 107M-1 and 5.0 × 108M-1, respectively. Epitope analysis showed that the two McAbs directed against the same epitope