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.Andtheirunderstandingoft

2、heimmuneresponsewaslimitedtowhattheycouldseeand,basedonthat,whattheycouldhypothesize.BeforeNowWhatisemergingisamorecomplexunderstandingofthesciencethatinvolvesmanyfieldsofscience.MolecularBiologyBiochemistryStructuralBiologyGenomicsProteomicsTransgenicandknockouta

3、nimalmodelsTechnologybasedonImmunologyTechnologywhichboostimmunologyAntibodiesFACS(Analysisandsorting)ELISAELISPOTTetramercellculture(invitroculturesystem)GeneProfiling（parallelsequencing）ProteomicsTransgenicandknockoutanimalsTechnologybasedonImmunologyTechnologyw

4、hichboostimmunology1.AntibodiesRecogniseandbindtomolecules(antigens)onforeignparticles,markingthemfordestruction.Eachantigenmaygenerateseveralantibodiesfordifferentsites(epitopes)onantigen.ProteinssecretedbyB-lymphocytes(typeofwhitebloodcell),invertebrates.170s

5、uppliersinUSPolyclonalantibodyAntigenImmunizationBleedingMonoclonalantibodyFreezeAbpurification*AffinityPurification:ProteinA/G,Antigenpeptideconjugatedonsepharoseoragarosebeads.ProteinAisfromthecellwallofS.aureus，MW42kDaProteinGisfromthecellwallof-hemolyticstrept

6、ococci,Gstrain，MW30~35kDa.ProteinAorProteinGcanbindtotheFcdomainofAb.AbpurificationAntibodyPurificationAffinitychromatographyusesantigen-antibodybindingtopurifyantigensorantibodiesTopurifyaspecificantigenfromacomplexmixtureofmolecules,amonoclonalantibodyisattached

7、toaninsolublematrix,suchaschromatographybeads,andthemixtureofmoleculesispassedoverthematrix.Thespecificantibodybindstheantigenofinterest;othermoleculesarewashedaway.SpecificantigenisthenelutedbyalteringthepH,whichcanusuallydisruptantibody-antigenbonds.Antibodiesca

8、nbepurifiedinthesamewayonbeadscoupledtoantigen(notshown).AffinitychromatographyAntibodyconjugationBeads,sepharose–TargetproteinpurificationFluorochrome–