抗人端粒重复序列结合因子(TRF1^(33-277))单克隆抗体的制备及生物学特性初步鉴定  

Preparation and characterization of monoclonal antibody against human telomeric repeat binding factor 1

作　　者：黄河[1] 施继敏[1] 陈巧芳[1] 罗依[1] 丁伟[1] 楼基余[1] 

机构地区：[1]浙江大学医学院附属第一医院,杭州310003

出　　处：《中华血液学杂志》2002年第12期631-633,共3页Chinese Journal of Hematology

基　　金：国家自然科学基金资助项目 (3 9870 3 3 9)

摘　　要：目的　制备抗人端粒重复序列结合因子片段 (TRF13 3 2 77)单克隆抗体 (单抗 ) ,探讨其生物学特性。方法　以GST TRF13 3 2 77融合蛋白为抗原 ,经细胞融合及ELISA方法筛选获得阳性杂交瘤细胞 ,对其进行染色体核型分析 ,将此单抗作为探针用于Westernblot和免疫组化检测标本。结果　获得1个抗TRF1蛋白的杂交瘤细胞株。以抗TRF13 3 2 77单抗作Westernblot检测正常人及急性白血病患者骨髓、大肠癌组织及近旁正常粘膜组织提取蛋白 ,在相对分子质量 6 0 0 0 0处有一特异性条带 ,与阳性对照纯化TRF1蛋白相一致 ,免疫组化显示该抗体在上皮细胞及骨髓造血细胞胞浆中阳性着色。结论　制备了具有高度特异性的抗TRF13 3 2 77单抗 ;建立了Westernblot和免疫组化检测组织表达TRF1蛋白的方法。Objective To prepare a monoclonal antibody against human telomeric repeat binding factor 1(TRF1) protein and explore its biological characteristics.Methods BALB/c mice were immunized with GST TRF1 33 277 fusion protein for the preparation of monoclonal antibody by hybridoma technique. The obtained antibody was used for clinical assay by Western blot and immunohistochemical staining. Results One strain of hybridoma was obtained. It was confirmed by Western blot that the antibody specifically recognized the 60 kD TRF1 protein. Immunohistochemical staining of the antibody showed that TRF1 protein located in the cytoplasm of epithelial cells and bone marrow cells. Conclusion A TRF1 monoclonal antibody,with high specificity was developed. It is useful for detection of TRF1 protein in tissue specimens.

关 键 词：生物学特性 鉴定 人端粒重复序列结合因子 单克隆抗体 免疫组化 制备 

分 类 号：R730.2[医药卫生—肿瘤]