抗人GCRG213单克隆抗体的制备与鉴定  

Preparation and identification of monoclonal antibody against human GCRG213

作　　者：伍银桥[1] 吴本俨[1] 王刚石[1] 尤纬缔[1] 王卫华[1] 王孟薇[1] 

机构地区：[1]解放军总医院南楼消化科,北京100853

出　　处：《细胞与分子免疫学杂志》2012年第1期46-48,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：军队"十一五"科研基金资助项目(06MA280);国家自然科学基金资助项目(81070296)

摘　　要：目的:制备GCRG213单克隆抗体(mAb)并进行初步鉴定。方法:在大肠杆菌中表达HIS-GCRG213融合蛋白,并以所获蛋白作为免疫原制备鼠mAb。采用ELISA、Westernblot法鉴定抗体的效价及特异性。免疫组化染色观察GCRG213在胃癌和正常组织中的表达。结果:HIS-GCRG213融合蛋白在大肠杆菌中获得高效表达,经常规的细胞融合和筛选获得2株可稳定分泌抗GCRG213的杂交瘤细胞株。ELISA法检测腹水的效价可达到1∶106,Western blot证实该抗体可与重组HIS-GCRG213蛋白特异性结合。免疫组化染色显示GCRG213在胃癌组织中的表达明显强于正常胃黏膜组织。结论:成功地制备出2株抗GCRG213的mAb,为进一步研究GCRG213的生物学功能提供了有效的工具。AIM： To prepare and characterize the monoclonal antibody against human GCRG213. METHODS： The HIS-GCRG213 fusion protein was expressed in E. coll. Mice were immunized with the purified HIS-GCRG213 protein. Hybridoma cell lines secreting monoclonal antibodies against GCRG213 were screened by regular cell fusion and subcloning approach. The titer and specificity of the antibody was characterized by ELISA and Western blot, re- spectively. The expression of GCRG213 was determined using immunohistochemistry technique on paraffin-embedded tissue sections from normal gastric mucosal tissues and advanced gastric cancer. RESULTS： The HIS-GCRG213 fusion protein with relative molecular mass of 20 800 was over expressed in E. coil Two hybridoma cell lines which secreted monoclonal antibody specifically against human GCRG213 fusion protein were successfully obtained. The ascite titers of this monoclonal antibody reached 1 ： 106. Western blot analysis showed that the monoclonal antibody could bind to the recombinant HIS-GCRG213 protein specif- ically. The immunohistochemistry showed that GCRG213 were expressed higher in gastric cancer tissues than in normal ones. CONCLUSION： The monoclonal antibody against human GCRG213 with high titer and specificity has been successfully prepared, which could be utilized as a useful reagent for further studying the biological function of the GCRG213.

关 键 词：胃肿瘤 GCRG213 单克隆抗体 

分 类 号：R392.11[医药卫生—免疫学]