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抗人凋亡相关蛋白TFAR19的单克隆抗体制备及鉴定
作者姓名:Chen Y  Zhang Y  Sun R  Song Q  Di C  Ma D
作者单位:北京大学基础医学院免疫学系,北京
基金项目: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蛋白在调亡细胞中高表达。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。

关 键 词:TFAR19  PDCD5  单克隆抗体  制备  鉴定

Preparation and identification of monoclonal antibodies against human apoptosis-related protein TFAR19
Chen Y,Zhang Y,Sun R,Song Q,Di C,Ma D.Preparation and identification of monoclonal antibodies against human apoptosis-related protein TFAR19[J].Acta Academiae Medicinae Sinicae,2000,22(6):502-504.
Authors:Chen Y  Zhang Y  Sun R  Song Q  Di C  Ma D
Institution:Department of Immunology, School of Basic Medical Sciences, Peking University, Beijing 100083, China. yingyu_chen@yahoo.com
Abstract:OBJECTIVE: 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 screened 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.
Keywords:TFAR19  PDCD5  monoclonal antibody  apoptosiT
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