一株鼠抗人PD-1单克隆抗体的制备及其生物学特性鉴定  

Preparation of a mouse anti-human PD-1 monoclonal antibody and identification of its biological characteristic

作　　者：曹莎莎[1] 徐永芳[1] 郑小翠[1] 葛彦[1] 居颂文[2] 居颂光[1] CAO Sha-sha XU Yong-fang ZHENG Xiao-cui GE Yan JU Song-wen JU Song-guang(Department of Immunology, School of Biology and Basic Medical Sciences, Soochow University, Suzhou Jiangsu 215123 Digestive Research Center, North District of Suzhou Municipal Hospital, Suzhou Jiangsu 215008,China)

机构地区：[1]苏州大学医学部免疫学系,江苏苏州215123 [2]苏州市立医院北区消化研究中心,江苏苏州215008

出　　处：《江苏大学学报（医学版）》2016年第1期17-20,共4页Journal of Jiangsu University：Medicine Edition

基　　金：国家自然科学基金资助项目(31370887;81373149;31200661);苏州市科技发展计划项目(SYS201363)

摘　　要：目的:制备鼠抗人PD-1单克隆抗体并对其生物学特性进行鉴定。方法:以稳定表达人PD-1分子的基因转染细胞株L929/PD-1免疫BALB/c小鼠,采用流式细胞术筛选分泌鼠抗人PD-1单抗的杂交瘤细胞株。用Ig G亚型快速定性试纸法、肿瘤细胞株表面PD-1分子识别谱检测、抗体效价测定、竞争结合抑制实验和蛋白质印迹等方法对所获PD-1单抗的生物学特性进行分析鉴定。结果:获得1株持续稳定分泌PD-1单抗的杂交瘤细胞株,命名为6E1。单抗6E1的重链为Ig G1、轻链为κ。单抗6E1对肿瘤细胞株U937、Thp-1、Jurkat细胞表面PD-1分子的识别谱与商品化PD-1单抗MIH4一致。单抗6E1与商品化抗体MIH4具有相似的识别位点。蛋白质印迹检测结果表明单抗6E1可特异性识别PD-1分子。结论:成功获得了1株鼠抗人PD-1单克隆抗体。Objective： To prepare mouse anti-human PD-1 monoclonal antibodies （mAbs） and identify its biological characteristics. Methods： BALB/c mice were immunized with the gene transfected cell line L929/PD-1. The anti-PD-1 mAb was identified with mouse IgG rapid isotyping analysis, recognition profile assay, antibody titer evaluation, competitive inhibition test and Western blotting. Results： A hybridoma cell line that secreting anti-PD-1 mAb was successfully obtained, named 6E1. The heavy chain of mAb was IgG1, and its light chain was K chain. It could detect PD-1 molecules expressed on U937, Thp-1 and Jur- kat cell lines, similar to the commercialization PD-1 mAb （MIH4）. The competitive inhibition assay showed that mAb 6E1 bind to the similar epitope with mAb MIH4. Western blotting results demonstrated that 6E1 could recognize PD-1 protein specificity. Conclusion： Mouse anti-human PD-1 monoclonal anti- body was successfully established.

关 键 词：PD-1 PD-L1 单克隆抗体 

分 类 号：R392.1[医药卫生—免疫学]