抑制和激活蛋白1(Rap1)单克隆抗体的制备和鉴定  

Preparation and identification of monoclonal antibody against repression and activation protein 1

作　　者：孟麟[1] 寿成超[1] 廉沈沂[1] 

机构地区：[1]北京大学肿瘤医院暨北京市肿瘤防治研究所,生物化学与分子生物学实验室,恶性肿瘤发病机制及转化研究教育部重点实验室

出　　处：《细胞与分子免疫学杂志》2015年第3期397-401,共5页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金青年项目(81301747)

摘　　要：目的制备并鉴定人端粒相关蛋白抑制和激活蛋白1(Rap1)的单克隆抗体。方法用重组原核人Rap1蛋白免疫BALB/c小鼠;细胞融合后,间接ELISA筛选阳性杂交瘤,建立分泌抗人Rap1蛋白单克隆抗体(m Ab)的杂交瘤细胞株,并进行效价测定和特异性鉴定。结果制备并获得1株稳定分泌抗人Rap1蛋白m Ab的杂交瘤细胞株。ELISA测定腹水效价高达1∶10 000以上。Western blot法、免疫沉淀和免疫荧光染色证实该m Ab能特异性识别人Rap1蛋白并可用于上述不同检测。结论成功制备获得了亲和力高、特异性好的抗人端粒相关蛋白Rap1的m Ab。Objective To prepare and identify monoclonal antibody （rnAb） against human telomere-associated repression and activation protein 1 （Rap1）. Methods BALB/c mice were immunized with recombinant Rap1 protein. The spleen ceils of the immunized mice were fused with Sp2,/0 cells. The positive clones were confirmed and selected by indirect ELISA for titer determination and specificity identification. Results One hybridoma cell strain secreting specific mAb against Rapl was obtained, namely 6F11. ELISA showed that the titer of the ascites was 1：10 000. Western blotting, immunoprecipitation and immunofluorescence experiments demonstrated that the mAb could specifically recognize and bind Rap1. Conclusion We have prepared mAb against human telomere-associated protein Rap1. The Rapl mAb has a good binding ability and specificity.

关 键 词：抑制和激活蛋白1(Rap1) 单克隆抗体 鉴定 

分 类 号：Q813.2[生物学—生物工程] R392.11[医药卫生—免疫学]