抗银杏二萜内酯葡胺注射液蛋白的单克隆抗体制备

Preparation of monoclonal antibody against proteins of ginkgo diterpene lactones meglumine injection

目的制备抗银杏二萜内酯葡胺注射液蛋白的单克隆抗体。方法将银杏叶总蛋白进行二维电泳,取蛋白点进行质谱分析并合成多肽。利用多肽作为免疫原免疫BALB/c小鼠,取其脾细胞与SP2/0细胞进行融合,通过间接ELISA方法筛选抗银杏二萜内酯葡胺注射液蛋白的单克隆抗体。结果 5株单克隆抗体亚类鉴定均为IgM,轻链为Kappa链。Western blotting实验证实单抗均能与银杏叶蛋白反应。结论单克隆抗体的制备为快速检测银杏二萜内酯葡胺注射液蛋白的方法建立奠定了基础。

Objective To obtain the monoclonal antibodies against the protein of ginkgo diterpene lactones meglumine injection. Methods After total proteins of ginkgo leaf running two-dimensional electrophoresis, the protein spots were digged and analyzed by mass spectrometry （MS）. According to the result of MS, the polypeptide was synthesized and used to immunize the BALB/c mice. The indirect ELISA was utilized to develop monoclonal antibody after cell fusion between SP2/0 cells and spleen cells from the immunized BALB/c mice. Results The immunoglobulin subtypes of five monoclonal antibodies were IgM and light chain was Kappa identified with a commercial capture-ELISA kit. Western blotting analysis also indicated that the monoclonal antibodies were specific to the protein of ginkgo leaf. Conclusion The experiment would be helpful for the establishment of detection method specific to the proteins of ginkgo diterpene lactones meglumine injection in the future study.