BTN3A3单克隆抗体的制备与鉴定

摘要

目的 制备BTN3A3单克隆抗体并鉴定.方法 用含小鼠IgG2a Fc标签的BTN3A3重组蛋白BTN3A3-mFc作为抗原免疫BALB/c小鼠,通过细胞融合获得杂交瘤细胞.用含人IgG1 Fc标签的BTN3A3重组蛋白BTN3A3-hFc作为检测原,筛选分泌BTN3A3单抗的杂交瘤细胞株.利用Western印迹法和间接免疫荧光对上述单抗进行鉴定.结果 共获得7株可稳定分泌BTN3A3单抗的杂交瘤细胞株.ELISA、Western印迹法和间接免疫荧光结果显示上述抗体可特异性地识别BTN3A3.结论 获得可应用于ELISA、Western印迹法及免疫荧光的BTN3A3单抗,为BTN3A3的功能性研究奠定了基础.%Objective To prepare and identify monoclonal antibodies against BTN 3A3.Methods BALB/c mice were immunized with the BTN3A3 recombinant protein BTN3A3-mFc containing the mouse IgG2a Fc tag as an antigen, and hybridoma cells were obtained by cell fusion .BTN3A3 recombinant protein BTN3A3-hFc containing a human IgG1 Fc tag as a detection target was used to screen hybridoma cell lines secreting monoclonal antibodies against BTN 3A3.The above monoclonal antibodies were identified with Western blot and indirect immunofluorescence .Results Seven hybridoma cell lines that could stably secret BTN 3A3 monoclonal antibodies were obtained . ELISA, Western blot and indirect immunofluorescence showed that the above antibodies specifically recognized BTN 3A3.Conclusion BTN3A3 monoclonal antibodies that can be applied to ELISA , Western blot or immunofluorescence have been obtained , which can contribute to the functional study of BTN3A3.