浙大 高级免疫学技术 英文课件

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1、免疫学相关技术（TechnologiesinImmunology）王迪浙江大学免疫学研究所wangdi80@gmail.comYearsago,immunologiststypicallyspentthebulkoftheirtimeatthelaboratorybench.Theirresearchinvolvedpeeringintoamicroscopeandprobablycharacterizingthedifferentcellsfromabloodsample.Andtheirunderstandingoftheimmuneresponsewaslimitedto

2、whattheycouldseeand,basedonthat,whattheycouldhypothesize.BeforeNowWhatisemergingisamorecomplexunderstandingofthesciencethatinvolvesmanyfieldsofscience.MolecularBiologyBiochemistryStructuralBiologyGenomicsProteomicsTransgenicandknockoutanimalmodelsTechnologybasedonImmunologyTechnologywhichboo

3、stimmunologyAntibodiesFACS(Analysisandsorting)ELISAELISPOTTetramercellculture(invitroculturesystem)GeneProfiling（parallelsequencing）ProteomicsTransgenicandknockoutanimalsTechnologybasedonImmunologyTechnologywhichboostimmunology1.AntibodiesRecogniseandbindtomolecules(antigens)onforeignpartic

4、les,markingthemfordestruction.Eachantigenmaygenerateseveralantibodiesfordifferentsites(epitopes)onantigen.ProteinssecretedbyB-lymphocytes(typeofwhitebloodcell),invertebrates.170suppliersinUSPolyclonalantibodyAntigenImmunizationBleedingMonoclonalantibodyFreezeAbpurification*AffinityPurifica

5、tion:ProteinA/G,Antigenpeptideconjugatedonsepharoseoragarosebeads.ProteinAisfromthecellwallofS.aureus，MW42kDaProteinGisfromthecellwallof-hemolyticstreptococci,Gstrain，MW30~35kDa.ProteinAorProteinGcanbindtotheFcdomainofAb.AbpurificationAntibodyPurificationAffinitychromatographyusesantigen-ant

6、ibodybindingtopurifyantigensorantibodiesTopurifyaspecificantigenfromacomplexmixtureofmolecules,amonoclonalantibodyisattachedtoaninsolublematrix,suchaschromatographybeads,andthemixtureofmoleculesispassedoverthematrix.Thespecificantibodybindstheantigenofinterest;othermoleculesarewashedaway.Spe

7、cificantigenisthenelutedbyalteringthepH,whichcanusuallydisruptantibody-antigenbonds.Antibodiescanbepurifiedinthesamewayonbeadscoupledtoantigen(notshown).AffinitychromatographyAntibodyconjugationBeads,sepharose–TargetproteinpurificationFluorochrome–