Protein tech-PPT 20110425

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1、ProteomicstechnologybaseonMSDept.ofBioMedicalResearchbgi-med@genomics.cnApr.2011Proteomicspropertiessample：morecomplex,noamplificationinstrument：tandemmassspectrometrybioinformatics：basedondatabaseClassicalproteomicspipelineSamplepreparationseparatio

2、nMSanalysisbioinformatics©JuergenCox,MPIBiochemistrySeparationtechnologybasedon2D-PAGE→highthroughputproteomicsbasedonLCLC2DEFunctionalitiesofLC(liquidchromatography)IntensityIntensitypeptideapeptidebpeptidecpeptideaMinMinpeptidebpeptidecTimeTimePeptide

3、solutiongoingthroughatubePeptidesolutiongoingthroughatubewithoutcolumnmaterialwithcolumnmaterialIdeasbyLiangPengIllustrationofthefunctionalitiesofLC,separationandenrichment(Ministhe(imaginary)minimalintensitythatcanbedetectedbymassspec)5MUDIPmassspectro

4、metrymassspectrometryinletIonsourceAnalyzerDetector1.IontrapIT1.GC1.ESI2.TimeofflightTOF2.LC2.MALDI3.quadrupoleQTandemMSGenerationofMSandMS/MSspectra©JimmyEng,ISBMassspecgeneratesMSandMSspectrainintertwiningtimepoint.10BGIMSPlatformQTRAPOrbitrapvelosmaX

5、isultraflexProducerABSCIEXThermoScientificBrukerBrukerSource-ESI-tripleQ-ESI-LTQ-OrbitrapESI-Q-TOF-MSMALDI-TOF-TOFAnalyzerLITResolution＞12,000＞100,000＞60,000≥40,000Accuracy≤700ppm<2ppm<5ppm≤3ppm•QCApplicationMRMHighaccuracyMSiTRAQ•MSimageQTRAP5500,ABSCI

6、EXOrbitrapvelos,ThermoScientificmaXisQ-TOF,BrukerultrafleX,BrukerThesearchingengineschema:MarkWDuncan,RuediAebersold&RichardMCaprioli，NatureBiotechnologyVolume:28,Pages:659–664Yearpublished:(2010)ProteomeprofilingQuantitativeproteomicsModificationproteo

7、micsTargetproteomicsProjectprocessReceivedsample,firstfundinitiateprojectSamplepreparationproteinextractionandtest-BrandforddataandSDS-PAGEfigureenzymolysisandseparationsteps.MSanalysisBioinformaticsanalysisProjectreportProteomeprofilingProteinprof

8、ilingstudyaimstolaunchmaximumamountofproteinidentificationfromthecomplexsampleextract,includingtissue,plasma,bodyfluidetc.Itcanprovidenecessaryfundamentalknowledgeforproteomicsanalysis.15workflowProteinExperiment：Bioinformaticsan