抗人凋亡相关蛋白TFAR19的单克隆抗体制备及鉴定  

Preparation and Identification of Monoclonal Antibodies against Human Apoptosis- related Protein TFAR19

作　　者：陈英玉[1] 张颖妹[1] 孙荣华[1] 宋泉声[1] 狄春辉[1] 马大龙[1] 

机构地区：[1]北京大学基础医学院免疫学系,北京100083

出　　处：《中国医学科学院学报》2000年第6期502-504,共3页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金!( 39870427)

摘　　要：目的研制鼠抗人凋亡相关蛋白 TFAR19的单克隆抗体，以进一步探讨 TFAR19蛋白的结构与功能。方法以纯化的重组人 TFAR19蛋白为抗原 ,免疫 BALB/c小鼠，将 TFAR19蛋白免疫的小鼠脾细胞与小鼠骨髓瘤细胞 SP2/0进行细胞融合，经克隆筛选获得鼠抗人 TFAR19蛋白单抗的杂交瘤细胞株，以间接 ELISA、 Western免疫印迹分析等方法进行单克隆抗体的鉴定。结果获得了 3株稳定分泌抗人 TFAR19单克隆抗体的杂交瘤细胞株 (C1,C10,2C12),其分泌的单抗效价高，特异性好，亲和力不一，免疫球蛋白亚类均为 IgG1。 Western免疫印迹分析证明 TFAR19蛋白在凋亡细胞中高表达。结论所获单抗能特异识别原核及真核细胞表达的 TFAR19蛋白，为进一步探讨 TFAR19蛋白的生物学效应提供了条件。To obtain mouse anti- human TFAR19 monoclonal antibodies for further study of the structure and function of the apoptosis- related protein TFAR19. Methods BALB/c mice were immunized with recombinant human TFAR19, hybridoma cells were secreened by cell fusion and subcloning approach. The monoclonal antibodies were identified by ELISA and Western blotting. Results Three hybridoma cells (C1， C10， 2C12） stable in secreting anti- TFAR19 monoclonal antibodies were obtained. The monoclonal antibodies showed high specificity and high titer to TFAR19 with various affinity. All of them belong to IgG1 subclass. Conclusions These monoclonal antibodies could bind specifically to TFAR19 protein.

关 键 词：TFAR19 PDCD5 单克隆抗体 制备 鉴定 

分 类 号：R392－33[医药卫生—免疫学]