单克隆抗体-沈威

《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