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1、第21卷3期中国病毒学21（3）：207-2122006年5月VIROLOGICASINICAMay2006抗SARS-CoVN蛋白单克隆抗体的特异性及其*识别抗原表位的初步鉴定1**1111王健伟，王彦斌，常昭瑞，闫克夏，谭文杰，12**11屈建国，夏宁邵，阮力，洪涛（1.中国疾病预防控制中心病毒病预防控制所,北京100052；2.福建省医学分子病毒学研究中心，厦门大学细胞生物学与肿瘤细胞工程教育部重点实验室,厦门361005）SpecificityandEpitopeMappingofFourMonoclonalAntibodiesagainstSARS
2、-CoVNucleocapsidProtein1*1111WANGJian-wei，WANGYan-bin，CHANGZhao-rui，YANKe-xia，TANWen-jie，12*11QUJian-guo，XIANing-shao，RUANLi，HUNGTao(1.NationalInstituteforViralDiseaseControlandPrevention,ChineseCenterforDiseaseControlandPrevention,Beijing100052,China；2.TheResearchCenterforMolecula
3、rVirologyofFujianProvince,TheKeyLaboratoryofMinistryofEducationforCellBiologyandTumorCellEngineering,XiamenUniversity,Xiamen361005,China)Abstract:Inordertocharacterizethespecificityofthemonoclonalantibodies(McAbs)againstSARS-associatedcoronavirus(SARS-CoV)nucleocapsidproteinandtoid
4、entifytheepitopesrecognizedbytheMcAbs，thenucleocapsidproteinsofhumancoronavirusOC43(HCoV-OC43)and229E(HCoV-229E)wasexpressedinE.coli.ThespecificitiesoffourMcAbs(1-1C2,2-2E5,1-1D6,and2-8F11)wereexaminedbyWesternblottingaswellasindirectfluorescenceassay.Twelvedifferentrecombinanttrun
5、catedNproteinswerethenusedtoidentifytheepitopesrecognizedbytheMcAbsbyWesternblotting.Theresultsshowedthat:(1)McAbs1-1C2,2-2E5,and1-1D6recognizedneitherhumancoronvirusesHCoV-OC43andHCoV-229Enortheirnucleocapsidproteins,implyingapossiblespecificityofthese3McAbstoSARS-CoV;(2)Theepitop
6、esrecognizedbyMcAbs2-8F11,2-2E5,and1-1D6werelocatedbetweenthe30thand60thaminoacid(a.a.)residuesoftheSARS-CoVNprotein,whiletheepitopesrecognizedbytheMcAb1-1C2werelocatedbetween170thand184tha.a.residues.TheidentificationofthespecificepitopesofSARS-CoVNproteinisparamountfortheimmunolo
7、gicalcharacterization,thedevelopmentofaccurateimmunologicalassayaswellasforexploringthepathogenesisofSARS-CoV.Keywords：SARS-associatedcoronavirus(SARS-CoV);Nucleocapsidprotein;Monoclonalantibody;Epitope摘要：为了明确抗SARS-CoVN蛋白单克隆抗体的特异性，并鉴定其识别表位，首先在E.coli中表达了人类冠状病毒229E（HCoV-229E）和OC43（HC
8、oV-OC4）N蛋白，用Westernblottin
