日本血吸虫重组信号蛋白14-3-3的纯化及单克隆抗体的制备  

Purification of recombinant Schistosoma japonicum signaling protein 14-3-3 and the monoclonal antibody preparation

作　　者：曹亮[1] 王弦[2] 谢小强[2] 徐振山[3] 宋礼华[3] 

机构地区：[1]安徽大学生命科学学院,合肥230039 [2]安徽医科大学生物化学与分子生物学教研室,合肥230032 [3]安徽安科生物工程(集团)股份有限公司,合肥230088

出　　处：《生物学杂志》2009年第2期73-75,共3页Journal of Biology

基　　金：国家高技术研究发展计划(863计划)资助项目(编号:2004AA2Z3570)

摘　　要：纯化日本血吸虫(中国大陆株)重组信号蛋白(rSj14-3-3),并制备其单克隆抗体。以纯化后的rSj14-3-3蛋白为抗原免疫Balb/c小鼠,用杂交瘤技术制备抗rSj14-3-3的单克隆抗体,并通过ELISA方法和Western blotting测定抗体的效价与特异性。获得了大量高纯度的rSj14-3-3蛋白;筛选出了能够稳定分泌抗rSj14-3-3单抗的杂交瘤细胞株3H6。单抗亚型为IgG1。实验依靠大肠杆菌表达系统高效表达了rSj14-3-3蛋白,并利用该蛋白制备了单克隆抗体,可用于今后血吸虫病免疫诊断的实验研究。To purify recombinant Schistosoma japonicurn （Chinese strain） signaling protein 14-3-3 and to prepare the monoclonal antibodies （McAbs） against rSj14-3-3, Balb/c mice were immunized with purified rSj14-3-3 protein. The anti-rSj14-3-3 monoclonal antiody was obtained by using hybridoma technique. Characterization of the antibodies was performed by enzyme linked immunosorbent as- say （ELISA） and Western blotting analysis. Results showed that high purified rSj14-3-3 protein and the hybridoma cell line 3H6 secreting monoclonal antibodies （McAbs） against rSj14-3-3 were obtained. The subclass of McAbs was found to be IgG1. The rSj14-3-3 protein is expressed in E. coli cells efficiently and its antibodies were obtained. They can be used in further study about immunodiagnostic methods of Schistosomiasis japonica.

关 键 词：日本血吸虫 rSj14—3—3蛋白 单克隆抗体 

分 类 号：Q785[生物学—分子生物学]