【摘要】 目的 制备抗胰岛素单克隆抗体(mAb)。方法 用重组人胰岛素原类似物免疫BALB/c小鼠制备免疫脾细胞，运用淋巴细胞杂交瘤技术将其与小鼠骨髓瘤细胞SP2/0融合。结果 建立了7株分泌抗胰岛素mAb的杂交瘤细胞系，命名为A、B、C、D、E、F和G；7株mAb的Ig亚类分别为IgM、IgM、IgM、IgG2b、IgG1、IgG1和IgG1；在夹心ELISA试验中，7株mAb均能与人、牛和猪胰岛素发生特异性免疫反应；在免疫印迹试验中，7株mAb均能与重组人胰岛素原类似物结合呈现一条蛋白印迹带。结论 7株mAb对人、牛和猪胰岛素均具有免疫反应性，可用于人和动物胰岛素的测定和纯化。

    【关键词】 单克隆抗体(mAb)；胰岛素；重组人胰岛素原类似物

    Development of monoclonal antibodies against insulin by immunization with recombinant human proinsulin analog

    LIU Yu, WANG Yong-shan, SUN Mao-hua, et al.

    Center for Disease Control and Prevention of Nanjing Command, Military Medical Institute of Nanjing Command, Nanjing 210002, China

    【Abstract】 Objective To prepare monoclonal antibody (mAb) against insulinMethods Spleen cells from BALB/c mice immunized with recombinant human proinsulin analog were fused with SP2/0 myeloma cells by using hybridoma techniques.Results Seven hybridomas secreting mAb against insulin were established and designated A, B, C, D, E, F and G. The subclass of the seven mAb was IgM, IgM, IgM, IgG2b, IgG1, IgG1 and IgG1 respectively. The seven mAb could specifically react with insulin derived from human, cattle or pig in sandwich ELISA. The specific reaction of each mAb with recombinant human proinsulin analog was confermed by immunoblotting assay, only one stained protein band appeared on the transferred film.Conclusion The seven mAb are immunological specific to insulin derived from human, cattle or swine, and are available to the detection and purification of insulin. ......