预览加载中，请您耐心等待几秒...

蓝舌病病毒1型和16型VP2蛋白单克隆抗体的制备及B细胞表位的筛选.docx

预览
1/2
2/2

在线预览结束，喜欢就下载吧，查找使用更方便

下载提示 文本预览

如果您无法下载资料，请参考说明：

1、部分资料下载需要金币，请确保您的账户上有足够的金币

2、已购买过的文档，再次下载不重复扣费

3、资料包下载后请先用软件解压，在使用对应软件打开

蓝舌病病毒1型和16型VP2蛋白单克隆抗体的制备及B细胞表位的筛选 Title:PreparationofMonoclonalAntibodiesagainstBluetongueVirusSerotype1and16VP2ProteinandScreeningofB-CellEpitopes Abstract: Bluetonguediseaseisamajorconcernforlivestockindustryworldwide.Thecausativeagent,bluetonguevirus(BTV),hasmultipleserotypes,withtype1and16beingthemostprevalent.TheVP2proteinofBTVplaysacrucialroleinvirusinfectivityandisoftentargetedforserologicaldiagnosticsandvaccinedevelopment.Thispaperaimstodescribethepreparationofmonoclonalantibodies(mAbs)againstBTVserotype1and16VP2protein,aswellasthescreeningofB-cellepitopesonVP2. Introduction: Bluetonguedisease,causedbybluetonguevirus,affectsruminantsworldwide,resultinginsignificanteconomiclosses.BTVhasmorethan27serotypes,withserotype1and16causingwidespreadinfections.TheVP2proteinofBTVishighlyimmunogenicandisakeytargetforserologicdiagnosisandvaccinedevelopment.MonoclonalantibodiesagainstspecificserotypesandidentificationofB-cellepitopescanprovidevaluabletoolsfordiseasediagnosisandvaccinedesign. Methods: 1.ProductionofrecombinantVP2proteins:TheVP2genesofBTVserotype1and16wereclonedintoanexpressionvectorandexpressedinE.coli.Purifiedrecombinantproteinswereusedasantigensinsubsequentimmunizationandscreeningexperiments. 2.Immunizationofmice:BALB/cmicewereimmunizedwithpurifiedVP2proteinstoinduceanimmuneresponse.Freund'sadjuvantwasusedtoenhancetheimmuneresponse.ImmunizationwasperformedatregularintervalstoensuretheproductionofspecificantibodiesagainstVP2. 3.Cellfusionandgenerationofhybridomas:Spleencellsfromimmunizedmicewerefusedwithmyelomacellstogeneratehybridomas.HybridomaswerescreenedfortheproductionofVP2-specificantibodiesusingELISAandimmunofluorescenceassays. 4.CloningandexpansionofVP2-specifichybridomacells:Positivehybridomacellswereclonedbylimitingdilutiontoobtainmonoclonalantibody-producingcells.Theselectedhybridomaswereexpandedandmaintainedforfurthercharacterization. 5.ScreeningofB-cellepitopes:epitopemappingwascarriedoutusingoverlappingsyntheticpeptidesspanningtheentireVP2pro