日本血吸虫重组蛋白SjGST的表达纯化及单克隆抗体的制备和特性鉴定
Purification of recombinant Schistosoma japonicum protein SjGST and preparation and characterization of the monoclonal antibody
-
摘要: 目的:获取高纯度的日本血吸虫重组GST蛋白(rSjGST),并制备单克隆抗体。方法:纯化rSjGST后,以其作为抗原,免疫Balb/c雌鼠,并用杂交瘤技术制备抗rSjGST的单克隆抗体,以ELISA方法测定抗体的效价,以蛋白质免疫印迹法测定抗体的特异性。结果:纯化出大量的高纯度rSjGST,并且筛选出能够稳定分泌抗rSjGST单抗的杂交瘤细胞株3C12。用试剂盒检测出单抗的亚型为IgG1。结论:依靠大肠埃希菌表达系统,高效表达出rSjGST,成功制备单克隆抗体,为血吸虫病免疫诊断提供了研究基础。Abstract: Objective: To isolate and purify recombinant Schistosoma japonicum (Chinese strain) protein glutathione S-transferase (rSjGST) for preparing the monoclonal antibodies (McAbs) against rSjGST.Methods: Balb/c mice were immunized with purified rSjGST protein.The anti-rSjGST monoclonal antibody was obtained through hybridoma technique.Characterization of the antibody was performed by ELISA and Western blot analysis.Results: The data had shown that high purified rSjGST protein and the hybridoma cell line 3C12 secreting McAbs against rSjGST protein were obtained.The subclass of McAbs was confirmed to be IgG1.Conclusions: The rSjGST protein was induced to express by isopropy-β-D-thiogalactoside in prokaryotic hosts Escherichia coli with high efficiency and its antibodies were harvested.They can be applied to further study on immunodiagnostic methods of Schistosoma japonicum.
下载: