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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 prepare monoclonal antibodies against ginkgo diterpene lactam meglumine injection protein. Methods Two-dimensional gel electrophoresis of Ginkgo biloba total protein was carried out. Protein spots were analyzed by mass spectrometry and peptides were synthesized. BALB / c mice were immunized with the polypeptide as immunogen. The spleen cells were fused with SP2 / 0 cells and the monoclonal antibodies against ginkgo diterpene lactone meglumine injection were screened by indirect ELISA. Results The five monoclonal antibody subtypes were identified as IgM and the light chain as Kappa chain. Western blotting experiments confirmed that monoclonal antibody can react with ginkgo leaf protein. Conclusion The preparation of monoclonal antibodies provides a basis for the rapid determination of ginkgo diterpene lactam meglumine injection protein.