抗人DR5功能性抗体的研制  

Characterization of a novel agonstic anit-human death receptor5 monoclonal antibody

作　　者：刘广超[1] 李淑莲[1] 卢峰[1] 白慧玲[1] 张军[1] 杜耀武[1] 赵粤萍[1] 都景芳[1] 马远方[1] 

机构地区：[1]河南大学细胞与分子免疫学重点实验室

出　　处：《河南大学学报（医学版）》2007年第1期16-20,共5页Journal of Henan University:Medical Science

基　　金：河南省杰出人才创新基金项目(0321001800);河南省医学科技创新人才工程项目(2002119)

摘　　要：目的:研制抗人DR5功能性单克隆抗体。方法:采用杂交瘤技术制备抗人DR5的mAb;MTT方法筛选分泌有细胞毒活性mAb的杂交瘤细胞;亲和层析方法纯化mAb;夹心ELISA法测定mAb亚型;Western blotting和斑点ELISA法检测mAb的抗原表位类型;间接ELISA法检测mAb的特异性;观察抗体诱导细胞凋亡的形态学特征。结果:获得1株合成分泌有细胞毒活性抗体的杂交瘤细胞,其分泌的mAb命名为mDRA-6,为Ig G1,其抗原表位类型为构象表位,其特异性识别hDR5,与hFas、hDR4等无交叉反应。mDRA-6对Jurkat细胞具有细胞毒作用,经mDRA-6处理后,Jurkat细胞表现出细胞凋亡的形态学特征。结论:获得一种抗人DR5功能性抗体mDRA-6,在以TRAIL/DR5系统进行肿瘤治疗和探讨DR5的功能结构域研究方面具有广泛应用前景。Objective： To prepare anti-human DR5 monoclonal antibody （mAb） with apoptotic activity. Methods： mAb was prepared by hybridoma technique. Tumoricidal anti-human DR5 mAb was screened by MTT assay, mAb was purified by protein G affinity chromatography. IgG subclass was analyzed by Sandwich-ELISA. The antigen epitope type of mAb was determined by western blotting and Dot-ELISA. mAb speciality was analyzed by indirect-ELISA, the morphological changes of Jurkat cells treated with mDRA-6 were observed. Results： A hybridoma was obtained, which secrets anti mDR5 with intrinsic cytotoxicity named as mDRA-6, mDRA-6 belongs to IgG1. The epitope type was of conformation and mDRA-6 was specially identified as rDR5. Jurkat cells treated with mDRA-6 showed typic apoptostic morphological features, such as loss of membrane asymmetry, condensation of the cytoplasm and nucleus, and apoptotic body formation. Conclusion： An novel agonistic mAb against hDR5 has been prepared successfully, and it may be a useful tool in investigating tumor＇s therapy by using DR5 as target molecule and exploring the functional domain of DR5.

关 键 词：死亡受体5 细胞凋亡 单克隆抗体 TRAIL 

分 类 号：R392.11[医药卫生—免疫学]