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目的 制备Tropic180 8基因重组蛋白的单克隆抗体 ,并对其生物学特性进行鉴定。方法 用Tropic180 8基因重组蛋白作为抗原免疫BALB/C小鼠 ,取小鼠脾细胞与小鼠骨髓瘤 (SP2 / 0 )细胞融合 ,经ELISA筛选和有限稀释法获得分泌单克隆抗体的细胞株 ,WesternBlot等方法对其生物学特性进行鉴定。结果 获得 2株识别Tropic180 8基因重组蛋白的单克隆抗体的细胞株Ⅱ 4B、Ⅲ 4C。WesternBlot法显示该抗体特异性地识别Tropic180 8基因重组蛋白 ;ELISA法测定杂交瘤细胞培养上清及体内成瘤后产生的腹水的抗体效价分别为 1∶80、 1∶6 0 0 ;杂交瘤细胞染色体数平均为 90～ 10 0 ;亚类鉴定单抗的重链为小鼠IgG1,轻链为κ型。结论 成功地制备了Tropic180 8基因重组蛋白的单克隆抗体 ,为进一步研究Tropic180 8基因重组蛋白的功能提供了良好的基础。
Objective To prepare a monoclonal antibody against Tropic 180 8 gene and identify its biological characteristics. Methods BALB / C mice were immunized with the recombinant protein Tropic 180 8 as antigen and the mouse spleen cells were fused with mouse myeloma (SP2 / 0) cells. The cell lines secreting monoclonal antibodies were obtained by ELISA and limited dilution. Western Blot and other methods to identify their biological characteristics. Results Two strains of monoclonal antibodies that recognize Tropic1808 recombinant protein Ⅱ 4B, Ⅲ 4C were obtained. Western blotting showed that the antibody specifically recognized the recombinant protein Tropic1808, the antibody titers of the hybridoma cell culture supernatant and ascites produced by the tumor in vivo were 1:80 and 1: 600, respectively; The average number of chromosomes was 90 ~ 100; the heavy chain of subclass identification McAb was mouse IgG1, and the light chain was κ. Conclusion The monoclonal antibody of Tropic 180 8 recombinant protein was successfully prepared and provided a good foundation for further study on the function of Tropic 180 8 recombinant protein.