西尼罗病毒NS1蛋白特异性单克隆抗体的制备与鉴定  

Preparation and characterization of monoclonal antibodies against NS1 protein of West Nile virus

作　　者：马健男[1] 杨涛[1] 徐青元[1] 孙恩成[1] 林燕[1] 刘霓红[1] 杨银辉[2] 李文京[3] 步志高[1] 吴东来[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室农业部兽医公共卫生重点开放实验室,哈尔滨150001 [2]军事医学科学院微生物流行病研究所病原微生物生物安全国家重点实验室,北京100071 [3]中国动物疫病预防控制中心,北京100125

出　　处：《中国免疫学杂志》2011年第6期529-532,共4页Chinese Journal of Immunology

基　　金：国家高技术研究与发展计划(863)项目(No.2007AA02Z406);兽医生物技术国家重点实验室自主研究课题(SKLVBP201018)的资助

摘　　要：目的:制备针对西尼罗病毒NS1蛋白的特异性单克隆抗体。方法:以原核表达的重组NS1蛋白为免疫原,免疫BALB/c小鼠并通过常规杂交瘤技术将免疫后小鼠脾细胞与SP2/0细胞进行融合。以真核表达的重组NS1蛋白为检测用抗原,建立间接ELISA检测方法筛选分泌NS1蛋白单克隆抗体的杂交瘤细胞。并利用Western blot和IFA试验对所获得的单克隆抗体进行鉴定。结果:共获得了2株稳定分泌NS1蛋白单克隆抗体的杂交瘤细胞株,分别命名为WN-1C10和WN-3D10,其亚类鉴定分别属于IgG2a和IgG1。这2株单克隆抗体均能与西尼罗病毒NS1蛋白和病毒抗原发生特异性反应,而与日本脑炎病毒无交叉反应。结论:本实验成功制备出针对西尼罗病毒NS1蛋白的特异性单克隆抗体,为我国建立西尼罗病毒与日本脑炎病毒的血清学鉴别诊断方法奠定了基础。Objective：To prepare monoclonal anti/oodles （McAbs） against NS1 protein of West Nile virus （WNV-NS1）. Methods： BALB/c mice were immunized intraperitoneally with the puified NS1 protein expressed by prokaryotie expression system. Then the spleen cells of immunized mouse were fused with SP2/0 myeloma cells by the PEG regent. Meanwhile, an indirect ELISA using the purified NSI protein derived from eukaryotic expression system as antigen was developed to screen antiboxty-produeing hybridomas. Western blot and IFA were applied to characterize the McAbs. Results： Two hybridoma cell strains secreting McAbs against WNV-NS1 protein, designated as WN-1CI0 and WN3D10, were obtained. Conclusion：The results of Western blot and IFA indicate that these two McAbs are specific for WNV-NS1 protein, and don＇t cross-react with Japanese encephalitis virus （JEV）, so they are helpful to develop differential diagnosis between WNV and JEV.

关 键 词：西尼罗病毒 NS1蛋白 单克隆抗体 

分 类 号：R392.12[医药卫生—免疫学]