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ID:43486784
大小:794.00 KB
页数:22页
时间:2019-10-08
《单克隆抗体-沈威》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
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.抗体(Ig):(Immunoglobulin)抗体分类:IgG、IgA、IgM、IgD、IgE抗体结构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
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