hPPARγ2单克隆抗体的制备与鉴定  

Preparation and identification of hPPARγ2 monoclonal antibody

作　　者：李燕[1] 郝志勇[2] 陈晓光[1] 

机构地区：[1]中国医学科学院药物研究所,北京100050 [2]中国医学科学院基础医学研究所,北京100050

出　　处：《中国免疫学杂志》2010年第6期533-537,556,共6页Chinese Journal of Immunology

基　　金：国家自然科学基金;北京市自然科学基金资助项目

摘　　要：目的:制备鼠源抗hPPARγ2单克隆抗体,并对其特异性进行鉴定。方法:利用已建立的pMD18-T/hPPARγ2亚克隆扩增出hPPARγ2前1017bpcDNA。利用pET原核表达系统表达了含有hPPARγ2前339个氨基酸(C-Histag)融合蛋白,并进行了纯化。其次利用原核表达的重组hPPARγ2蛋白制备了特异性抗hPPARγ2单克隆抗体1B4、3H2、3H10、10D6和10D8。通过ELISA和Western blot及细胞免疫化学的方法对抗体进行了特异性鉴定。结果:成功制备了鼠抗hPPARγ2单克隆抗体,并证明这5株单抗均为hPPARγ2的特异性抗体。结论:制备的杂交瘤细胞株能稳定分泌高特异性的hPPARγ2抗体。这些系统的建立和鼠抗hPPARγ2单克隆抗体的制备为后续的针对hPPARγ2作用及机制研究提供了良好的工具。Objective：To prepare and identify the monoclonal antibody against hPPARγ2.Methods：cDNA of hPPARγ2 was obtained from pMD18-T/hPPARγ2.hPPARγ2 was expressed in prokaryocyte and purified successfully.The anti-hPPARγ2 monoclonal antibodies （mAbs） 1B4,3H2,3H10,10D6 and 10D8 were produced by immunization with purified hPPARγ2.The specificity of the antibodies was identified by ELISA,Western blot assay and immunocytochemistry.Results：Five novel murine monoclonal antibodies only against hPPARγ2 were harvested,and they were all specific to hPPARγ2.Conclusion：The antibodies obtained provide more useful tools for following research and settling the basis for screening new drugs and mechanism.

关 键 词：hPPARγ2 单克隆抗体 制备 特异性 

分 类 号：R392.33[医药卫生—免疫学]