小鼠抗人LSECtin单克隆抗体的制备与鉴定  

Preparation and characterization of mouse monoclonal antibody against human LSECtin

作　　者：彭贵林 刘静 柳迪 唐丽 PENG Gui-lin;LIU Jing;LIU Di;TANG Li(Institute of Lifeomics,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 102206,China;State Key Laboratory of Proteomics,Beijing Proteome Research Center,National Center for Protein Sciences(Beijing),Beijing 102206,China)

机构地区：[1]军事科学院军事医学研究院生命组学研究所,北京102206 [2]蛋白质组学国家重点实验室北京蛋白质组研究中心国家蛋白质科学中心(北京),北京102206

出　　处：《军事医学》2019年第6期431-435,共5页Military Medical Sciences

基　　金：国家自然科学基金面上项目（31570901）;北京市自然科学基金面上项目（7182035）;中国博士后科学基金面上项目（2018M633740）

摘　　要：目的制备并鉴定小鼠抗人LSECtin(肝、淋巴结窦内皮细胞凝集素)单克隆抗体,为深入研究其功能奠定基础。方法利用CHO细胞制备人LSECtin-His融合蛋白,将其作为抗原免疫BALB/c小鼠,通过电融合获得能产生人LSECtin单克隆抗体的杂交瘤细胞。用LSECtin重组蛋白作为检测原,筛选分泌LSECtin单克隆抗体的杂交瘤细胞株。分别通过流式检测和Western印迹实验(WB),对LSECtin单克隆抗体进行鉴定。结果获得分泌LSECtin单克隆抗体的杂交瘤细胞系。流式细胞术结果显示,共有5株抗体可特异性识别细胞表面表达的LSECtin;有2株可用于WB实验。结论获得了能应用于流式细胞术和WB实验的LSECtin单克隆抗体,为LSECtin的功能性研究奠定了基础。Objective To prepare and characterize mouse monoclonal antibodies against human LSECtin(liver and lymph node sinusoidal endothelial cells lectin)for further research on LSECtin.Methods Human LSECtin-his fusion protein was prepared using CHO cells and immunized into BALB/c mice.Hybridoma cell lines producing the LSECtin monoclonal antibodies,prepared using electrofusion,were screened by LSECtin recombinant protein.LSECtin monoclonal antibodies were characterized by flow detection and Western blotting(WB).Results Hybridoma cell lines that could stably secrete LSECtin monoclonal antibodies were obtained.Flow cytometry showed that the five antibodies could specifically recognize LSECtin expressed on the cell surface.Two antibodies could be applied to WB.Conclusion LSECtin monoclonal antibodies that can be applied to flow cytometry and WB have been obtained,which can facilitate the functional study of LSECtin.

关 键 词：LSECtin 杂交瘤细胞 单克隆抗体 CHO细胞 重组蛋白质类 流式细胞术 

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