抗人DR5单克隆抗体的制备、鉴定及活性分析  

Preparation,characterization,and activity analysis of monoclonal antibodies against human death receptor 5

作　　者：杨东海[1] 庄国洪[1] 刘忠臣[2] 张长弓[3] 何琼[3] 李文珠[1] 

机构地区：[1]厦门大学医学院抗癌研究中心 [2]厦门大学附属中山医院 [3]厦门市波生生物有限公司

出　　处：《免疫学杂志》2009年第6期671-675,共5页Immunological Journal

基　　金：福建省自然科学基金(C0710046)资助

摘　　要：目的制备抗人DR5单克隆抗体(mAb),鉴定其特性,并进行生物学活性分析。方法以纯化的可溶性DR5(sDR5)免疫Balb/c小鼠,杂交瘤技术制备抗人DR5mAb;运用ELISA、SDS-PAGE电泳方法测定抗DR5mAb与sDR5结合的特性;Ig亚类ELISA试剂盒鉴定抗DR5mAb亚类;间接ELISA法检测腹水mAb效价;流式细胞仪检测肿瘤细胞表面DR5的表达水平;流式细胞仪检测抗DR5单克隆抗体(mAb)诱导肿瘤细胞凋亡的功能。结果获得1株可分泌抗DR5mAb的杂交瘤细胞系R150。SDS-PAGE电泳检测证实,获得的R150可特异性地识别DR5;R150的Ig亚类为IgGI(λ型);腹水效价为1×106;通过流式细胞仪可敏感地检测到肿瘤细胞表面DR5的表达水平及R150诱导肿瘤细胞凋亡情况。结论获得1株可分泌抗DR5mAb的细胞系R150,抗体具有效价高、特异性强等特点并能有效诱导肿瘤细胞凋亡,具有较好的应用价值。Objective To prepare monoclonal antibodies (mAbs) against DR5,and characterize their biological properties by using ELISA,flow cytometry,and SDS-PAGE. Methods Balb/c mice were immunized with soluble DR5 (sDR5),and then mAbs was prepared by hybridoma technique. Ig subclass and specificity of mAbs were analyzed by ELISA and SDS-PAGE,respectively. The titres of mAbs in ascitic fluid were determined by indirect ELISA. Flow cytometry was utilized to detect the tumor cell apoptosis-inducing activity of the mAbs a...

关 键 词：死亡受体5 TRAIL 单克隆抗体 细胞凋亡 

分 类 号：R392[医药卫生—免疫学]