目的 制備抗銀杏二萜內(nèi)酯葡胺注射液蛋白的單克隆抗體。方法 將銀杏葉總蛋白進行二維電泳，取蛋白點進行質(zhì)譜分析并合成多肽。利用多肽作為免疫原免疫BALB/c小鼠，取其脾細(xì)胞與SP2/0細(xì)胞進行融合，通過間接ELISA方法篩選抗銀杏二萜內(nèi)酯葡胺注射液蛋白的單克隆抗體。結(jié)果 5株單克隆抗體亞類鑒定均為IgM，輕鏈為Kappa鏈。Western blotting實驗證實單抗均能與銀杏葉蛋白反應(yīng)。結(jié)論 單克隆抗體的制備為快速檢測銀杏二萜內(nèi)酯葡胺注射液蛋白的方法建立奠定了基礎(chǔ)。

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.

國家“重大新藥創(chuàng)制”——中藥治療心腦血管疾病有效成分組（群）的創(chuàng)新藥物孵化基地（2011ZX09401-097）