小鼠肝炎病毒N蛋白单克隆抗体的制备及鉴定  

Preparation and identification of monoclonal antibodies against recombinant N protein of mouse hepatitis virus

作　　者：周洁 赵莉[1,2,3] 胡建华 高诚 

机构地区：[1]上海实验动物研究中心 [2]上海市实验动物质量监督检验站 [3]中国科学院上海药物研究所,上海201203

出　　处：《细胞与分子免疫学杂志》2012年第8期841-843,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：上海市科委科技创新行动计划(08140901200)

摘　　要：目的:制备并鉴定小鼠肝炎病毒(MHV)N蛋白的单克隆抗体(mAb)。方法:以Bac-to-Bac杆状病毒表达系统表达的重组N蛋白为免疫原,免疫BALB/c小鼠,应用常规杂交瘤技术将免疫小鼠脾细胞与Sp2/0骨髓瘤进行融合,用间接ELISA方法筛选分泌N蛋白mAb的杂交瘤细胞系。并用Western blot和IFA方法对所获得的mAb进行鉴定。结果:共获得4株稳定分泌抗N蛋白mAb的杂交瘤细胞系,其亚类鉴定2株为IgG2a,1株为IgG2b,1株为IgG1,轻链均为κ型。经鉴定4株mAb均能与重组N蛋白发生特异性反应。结论:成功获得了特异性抗MHV N蛋白的mAb,为进一步研究N蛋白的结构功能及建立诊断学方法奠定了基础。AIM： To prepare and characterize the monoclonal antibodies（mAbs） against N protein of mouse hepatitis virus（MHV）.METHODS： BALB/c mice were immunostimulated with the purified recombinant N protein expressed by Bac-to-Bac baculovirus expression system.Then the spleen cells of the immunized mice were fused with Sp2/0 myeloma cells by conventional hybridoma techniques.An indirect ELISA using recombinant N protein as antigen was established to screen antibody-producing hybridoma cell lines.Western blotting and IFA were applied to characterize the mAbs.RESULTS： We obtained four hybridoma cell lines secreting mAbs against recombinant N protein.The subclasses of two mAbs were IgG2a,one IgG2b and the other IgG1,and light chains were both Kappa.Western blotting showed that the mAbs specifically recognized certain antigenic epitope of N protein of MHV.CONCLUSION： mAbs against N protein of MHV with high activity and specificity were prepared successfully,which provides a potential basis for further researches on N protein function and diagnostic methods.

关 键 词：小鼠肝炎病毒 重组N蛋白 单克隆抗体 

分 类 号：R392.11[医药卫生—免疫学] R373.21[医药卫生—基础医学]