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1、Thepreparationofmonoclonalantibodies生物1092沈威MonoclonalantibodiesMonoclonalantibodies(mAbormoAb)aremonospecificantibodiesthatarethesamebecausetheyaremadebyidenticalimmunecellsthatareallclonesofauniqueparentcell.AntibodyAnantibody,alsoknownasanimmunoglobulin,isalargeY-shapedproteinusedbytheimmunesyst
2、emtoidentifyandneutralizeforeignobjectssuchasbacteriaandviruses.Theantibodyrecognizesauniquepartoftheforeigntarget,termedanantigen.Eachtipofthe"Y"ofanantibodycontainsaparatope(astructureanalogoustoalock)thatisspecificforoneparticularepitope(similarlyanalogoustoakey)onanantigen,allowingthesetwostruc
3、turestobindtogetherwithprecision.抗體(Ig):（Immunoglobulin)抗體分類：IgG、IgA、IgM、IgD、IgE抗體結(jié)構(gòu)Cell-mediatedimmunityHumoralimmuneThebasicprincipleofmonoclonalantibodyNormallymphocytes(suchasmousespleencells)havetheabilitytosecreteantibodies，Butcannotlong-termculturedinvitro，Tumorcells(suchasmyeloma)canlong-te
4、rmculturedinvitro，Butcannotsecreteantibodies。GeorgesJ.F.KohlerCésarMilsteinTheproductiontechnologyofMonoclonalantibodies (Hybridomatechnology)Therearetwoadvantagestousecellfusiontechnologytogetmonoclonalantibodies:Lymphocytetumor:cancontinueProliferation，havenoabilitytoproducespecificantibodies。Lym
5、phocytes:canproducespecificantibodies，can'tcontinueProliferation。PreparationofmonoclonalantibodiesTheselectionandpreparationofCellSpleencells:formthemouse'sspleensuspensionMyelomacells:1、formthesamespecies2、highcelldensity3、inresting4、growfastlyABlymphocytes－－canproducespecificantibodyMousemyelomac
6、ells－－UnlimitedproliferationHybridcellgMethodofcellfusionPhysicalmethodsElectrofusionLaserfusionChemicalfusionBiologicalfusionFilterTheselectiveculturemediumiscalledHATmediumbecauseitcontainshypoxanthine,aminopterin,andthymidine.Thismediumisselectiveforfused(hybridoma)cells.Unfusedmyelomacellscanno
7、tgrowbecausetheylackHGPRT,andthuscannotreplicatetheirDNA.Unfusedspleencellscannotgrowindefinitelybecauseoftheirlimitedlifespan.Onlyfusedhybridcells,referredtoashybridomas,areabletogrowindefinitelyinthemedia
