國際肝臟蛋白質(zhì)組學(xué)研究

計劃的實施與毒理學(xué)發(fā)展機遇賀福初/孫啟鴻軍事醫(yī)學(xué)科學(xué)院復(fù)旦大學(xué)生物醫(yī)學(xué)研究院2005.09.21/沈陽報告提要蛋白質(zhì)組學(xué)產(chǎn)生的時代背景“人類蛋白質(zhì)組計劃”的目標(biāo)與意義蛋白質(zhì)組學(xué)與毒理學(xué)/環(huán)境醫(yī)學(xué)曼哈頓原子彈研制計劃人類基因組計劃阿波羅登月計劃人類歷史上的三大科技工程1941.12.6—1945.7.16羅斯福批準(zhǔn),耗資20億美元原子半徑10-10m原子體積10-30m3人體半徑100m人體體積100m3太陽系半徑1012m太陽系體積1034m31990.10.1-2003.4.23克林頓、布萊爾批準(zhǔn)，耗資30億美元1961.5.25—1969.7.20肯尼迪批準(zhǔn),耗資240億美元人類基因組計劃開創(chuàng)了

“基因組時代”基因組學(xué)向蛋白質(zhì)組學(xué)“求助”!

Nature409:747,15Feb.2001

Andnowforproteome

“現(xiàn)在輪到蛋白質(zhì)組”Science291:1221,16Feb.2001ProteomicsinGenomeland“基因組大地中的蛋白質(zhì)組學(xué)”PROTEIN轉(zhuǎn)錄組一種細(xì)胞、組織或生物體所對應(yīng)的全套mRNARNA基因組生物體所擁有的全套染色體上的全部基因DNA蛋白質(zhì)組一種細(xì)胞、組織或生物體所對應(yīng)的全套蛋白質(zhì)PROTEIN功能執(zhí)行體遺傳信息載體基因和蛋白質(zhì)并不存在嚴(yán)格的線性關(guān)系

ORF并不預(yù)示一定存在相對應(yīng)的功能性基因

mRNA水平并非與蛋白質(zhì)的表達(dá)水平對應(yīng)翻譯后修飾及同工蛋白質(zhì)(Isoforms)等現(xiàn)象在基因水平無從表現(xiàn)蛋白質(zhì)與蛋白質(zhì)的相互作用難以在基因水平得以認(rèn)識基因組與轉(zhuǎn)錄組不能取代蛋白質(zhì)組翻譯后修飾相互作用構(gòu)象變化翻譯后拼接移位胞質(zhì)胞核蛋白質(zhì)調(diào)節(jié)的多樣性生命的“萬花筒”蛋白質(zhì)功能的群體性繩墻扇茅蛇樹蛋白質(zhì)作用的整體性探索生命奧秘的直接對象蛋白質(zhì)組生命體的統(tǒng)一性源于基因組生命體的多樣性、復(fù)雜性、功能性、表型源于蛋白質(zhì)組1463-4x1043x109103(1012)

Proteomicsseekstoprovidefunctionalinformationforallproteins.proteomicsismoreofaconceptthanadefinedtechnology,anditreferstoanyprotein-basedapproachthathasthecapacitytoprovidenewinformationaboutproteins

onagenomewidescale.“Proteomicsincludesnotonlytheidentificationand

quantificationofproteinsbutalsothedeterminationof

their:

localization

modifications

interactions

activities

function報告提要蛋白質(zhì)組學(xué)產(chǎn)生的時代背景“國際人類肝臟蛋白質(zhì)組計劃”的目標(biāo)與意義蛋白質(zhì)組學(xué)與毒理學(xué)/環(huán)境醫(yī)學(xué)DNA序列圖≠基因圖人類基因組中1/3以上基因未曾確認(rèn)基因確認(rèn)的基本層次--蛋白質(zhì)水平天書解讀天書Science291:1221,2001全面揭示重大疾病發(fā)生發(fā)展機制的基礎(chǔ)蛋白質(zhì)組單基因病—遺傳病多基因病—腫瘤等

重大疾病發(fā)生發(fā)展機制單一基因單一蛋白基因群蛋白質(zhì)群轉(zhuǎn)錄組蛋白質(zhì)組????發(fā)現(xiàn)大量新型藥靶與新藥的源泉蛋白質(zhì)組最重要的疾病100-150每種疾病相關(guān)的基因10每個基因相關(guān)的蛋白質(zhì)3–10藥靶蛋白總數(shù)

3000-15,000

現(xiàn)有藥物約2000種85％是針對目前已知的500種藥靶6-30倍藥靶有待發(fā)現(xiàn)啟動人類蛋白質(zhì)組計劃勢在必行！探索人體生命奧秘的直接對象全面揭示重大疾病發(fā)生發(fā)展機制的基礎(chǔ)發(fā)現(xiàn)大量新型藥靶與新藥的源泉人類基因組序列及基因注釋人類蛋白質(zhì)組VIP:VeryImportantProteome人類蛋白質(zhì)組計劃的

主要科學(xué)目標(biāo)驗證人類基因組計劃推測的基因，注釋基因組闡明蛋白質(zhì)組的調(diào)控模式并與轉(zhuǎn)錄組進行對比建立蛋白質(zhì)群/組裝體，蛋白質(zhì)復(fù)合物或蛋白質(zhì)機器,即連鎖圖建立人體生理學(xué)、病理學(xué)的蛋白質(zhì)組基礎(chǔ)1463-4x1043x109103(1012)

基因組計劃蛋白質(zhì)組計劃Bodyfluidmostaccessibleforbiomarkerdiscoveryinanimalandhumans.LiverKidneyOrganBodyFluidBrainImmuneOrgans/VesselsIntestineOtherTissuesCSFFecesEndocrine/ParacrineSecretionsBileUrineAir/BALFLymphLungsEyesSkinOralCavitySweatSalivaTearsVenousBloodArterialBloodSerum10%90%SerumProteinsNon-InformativeAbundantProteinsi.e.albumin,IgG,etc.InformativeLowAbundanceProteinsEnrichmentStrategiesforInformativeSerumProteins:SELDI

SerumImmunosubtraction

人類肝臟蛋白質(zhì)組計劃里程碑蛋白表達(dá)譜蛋白連鎖圖亞細(xì)胞定位圖修飾譜人類基因組計劃

里程碑遺傳圖物理圖序列圖InitiativesofHumanProteomeProject(HPP)HPPPHumanPlasmaProteomeProject,USAHLPPHumanLiverProteomeProject,ChinaPSIProteomicsStandardsInitiativeUK

HBPP

HumanBrainProteomeProject,GermanyHAIHumanAntibodyInitiativeSwedenWhyisliver?wwwww.HLPP.HUPO為何研究肝臟?肝臟功能的多樣性與重要性能量——能量轉(zhuǎn)換、儲存物質(zhì)——代謝（活性分子的合成、毒性分子的分解）信息——信息分子的合成與分泌肝臟為其它組織提供能量—可氧化底物組織的活力依賴于高能鍵的連續(xù)生成。肝臟產(chǎn)生大部分脂肪酸，后者是禁食狀態(tài)下能量的基本來源。肝臟是給食狀態(tài)下由過剩糖合成脂肪酸的主要部位。藥物毒物營養(yǎng)物生物異源物生物轉(zhuǎn)化化學(xué)多樣性向體內(nèi)的生物轉(zhuǎn)化體系提出嚴(yán)峻挑戰(zhàn)1.2×107種以上的化學(xué)物(CAService’slist)以每周約8000種的速率增加常用化學(xué)物在63,000種以上大約11,500種，作為食物或藥物制劑添加物攝入其余的50,000種，為潛在的環(huán)境污染物人類發(fā)育過程中造血組織的興替造血系統(tǒng)的發(fā)育必需肝臟的“培育”合成大多數(shù)循環(huán)血漿蛋白合成和分泌血漿蛋白是肝臟實質(zhì)細(xì)胞——肝細(xì)胞的獨有功能（肝細(xì)胞占肝臟總細(xì)胞數(shù)的60%及肝臟質(zhì)量的約80%）最有特征的血漿蛋白是白蛋白，在大多數(shù)哺乳動物中占總血漿蛋白的55-60%凝血酶、抗凝因子、溶栓酶等肝臟再生機體再生能力最強的器官終生保持旺盛的再生能力肝臟中包含的“組”(-ome)Metabolicgenomics→Metabolome

(代謝組)Energygenomics→Energome

(能量組)Pharmacogenomics→Pharmacome

(藥理組)Toxicogenomics→Toxicome

(毒理組)Regeneratinggenomics

→Regenerome

(再生組)全球及中國肝炎的流行病學(xué)統(tǒng)計

全球:3.5億攜帶者中國:1.8億攜帶者死亡:23萬人/年疾病負(fù)擔(dān):500億元人民幣治療手段:抗病毒,保肝降 酶,抗纖維化,免疫治療等缺點:病毒易反彈,病 情易反復(fù)肝炎向肝癌的惡性轉(zhuǎn)化

難以遏制全世界現(xiàn)有3.5億慢性乙肝病毒（HBV）攜帶者，占世界人口的5%，亞洲和非洲HBV攜帶率為8-15%

HBV攜帶者中50-70%病毒復(fù)制活躍，為慢性乙肝

慢性乙肝病人肝硬化發(fā)生率為2-20%，代償性肝硬化發(fā)展成失代償性肝硬化為20-23%，發(fā)展成肝癌的占6-15%中國的慢性乙肝病人中，約25-40%最終將死于肝硬化或合并肝癌HBV攜帶者最終死于相關(guān)肝病的危險性，男性為50%，女性為15%

全球:100萬人/年中國:50萬人/年死亡:約45萬人/年疾病負(fù)擔(dān):400億元人民幣治療手段:手術(shù)切除(只有 15%-20%)治療效果:術(shù)后易轉(zhuǎn)移,易復(fù) 發(fā),五年存活率 40-50%全球及中國肝癌的流行病學(xué)統(tǒng)計原發(fā)性肝癌的治療水平亟待突破世界上每年有100萬新發(fā)原發(fā)性肝癌病人，其中40-50%在我國

我國原發(fā)性肝癌發(fā)病率和死亡率均位居第二位

近20年來我國肝癌的發(fā)病率上升近40%原發(fā)性肝癌一般起病較隱匿，早期缺乏典型臨床表現(xiàn)，初診大多已屬中晚期

初診肝癌病人中，只有15-20%的病人具備手術(shù)條件這些病人術(shù)后5年生存率僅為40-50%

肝癌是致死率最高的惡性腫瘤之一重大肝病的發(fā)生發(fā)展無法歸咎于少數(shù)幾個基因或蛋白質(zhì)所呈現(xiàn)的功能狀態(tài)和信號通路從蛋白質(zhì)組層面上“全景式”揭示肝臟疾病的生理病理機制是解決重大肝病的根本出路肝炎病毒肝臟/肝細(xì)胞肝炎肝硬化/纖維化原發(fā)肝癌肝癌轉(zhuǎn)移多因素、多步驟的發(fā)病機制肝臟是人類蛋白質(zhì)組計劃的首批目標(biāo)！Prometheusstolefirefromthegodsandgaveittomortals.PrometheusBoundHeraclesLiberatePrometheusHLPP—ModernPrometheusMythPrometheus

LiverEagleLiverdiseases

HeraclesHLPP人類肝臟蛋白質(zhì)組計劃

HumanLiverProteomeProject(HLPP)國際HLPP共同體Generateanintegrativeapproachleadingtoacomprehensivefunctionalmapoftheliver

Expandliverproteometoits“PHYSIOME”and“PATHOME”todramaticallyacceleratethedevelopmentofdiagnostics,preventionandtherapeuticstowardsitsdiseases

Developstandardoperatingprocedures(SOPs)forotherHUPOInitiativesVISIONHUPOWorkshop,Bethesda,April28-29,2002

BroadinterestsandsupportsforinitiatingHLPP

HUPOLiverProteomeWorkshop,Beijing,October22-24,2002

GettingconsensusviewforscientificobjectivesofHLPP

HUPO1stWorldCongress,Versailles,November21-24,2002

Co-chairsofHLPP:Drs.FuchuHe,JohnBergeronandChristianBréchot

HLPPPlanningCommittee:17members;May2003ProposalsaboutWorkingPlanofHLPP

Dr.JohnBergeron:Jan31,2003(Management);

Dr.FuchuHe:Feb17,2003(ScientificStrategy)Dr.FuchuHe:Mar22,2003(ActionPlan);

Dr.ChristianBrechot:Mar31,2003(Sampling)

HLPPOffice,March18,2003HUPOLPPWorkshop,Bethesda,July17-18,2003

NIHparticipatingHUPO2ndWorldCongress,Montreal,Oct.8-12,2003

Dr.FuchuHewasappointedastheexecutiveChairofHLPP(2003-2005)HUPOHLPPSatelliteMeeting,Montreal,Oct.12,2003SetupExecutiveCommittee&9Sub-committees,ActionPlanatPilotPhaseFrenchliversampleswerecollectedanddistributedamong8labs,May,2004HUPOHLPPSatelliteMeeting,Beijing,Oct.24,2004ChinesepartofHLPPwasofficiallylaunched,Nov.,2004Chineseliversampleswerecollectedanddistributedamong7labs,Feb.,2005MISION表達(dá)譜修飾譜定位圖連鎖圖樣品庫抗體庫數(shù)據(jù)庫HLPP的科學(xué)目標(biāo)---肝臟蛋白質(zhì)組的“太陽系”3-D圖ORF組肝臟蛋白質(zhì)組與

肝臟生理、病理的銜接肝臟蛋白質(zhì)組代謝組毒理組藥理組再生組肝炎組肝癌組能量組作為分泌器官，肝臟合成大多數(shù)的循環(huán)血漿蛋白

肝臟轉(zhuǎn)錄組“肝臟蛋白質(zhì)組計劃”和“血漿蛋白質(zhì)組計劃”的整合肝臟蛋白質(zhì)組血漿蛋白質(zhì)組?基因組肝臟蛋白質(zhì)組肝臟轉(zhuǎn)錄組基因組、轉(zhuǎn)錄組和蛋白質(zhì)組的集成證實推測基因?qū)Ρ日{(diào)控模式

重要功能蛋白質(zhì)肝病藥物靶標(biāo)肝病診斷標(biāo)志物人類肝臟蛋白質(zhì)組肝炎病毒肝臟/肝細(xì)胞感染肝炎肝硬化/纖維化原發(fā)肝癌肝癌轉(zhuǎn)移重大肝病預(yù)警、診斷、預(yù)防、治療的關(guān)鍵環(huán)節(jié)“東亞病夫”“肝病大國”1億多肝炎病毒攜帶者1000億多/年醫(yī)療費用SOPsSpecimenBankingPlatformsWorkingteamGlobalCollaborationExpressionProfileORFeome

AntibodiesBioinformatics…

ModificationProfileSubcellularLocalizationProtein-ProteinInteractionAntibodiesBioinformaticsIntegrateproteomewithtranscriptomeCompareproteomeswithinhealthanddiseasedliverTimeLinesPilotPhase(2003-2005)PhaseII(2006-2010)OutlineBackgroundProgressoftheHLPP-ProgressreportonexpressionprofilingInitiationoftheCNHLPPNextworkProgress-ExpressionprofilingEvaluationoftechnologiesBioinformaticsset-upSamplingandpreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissuesProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissuesMajorconclusionsfromthevaluationoftechnologies–16standardproteins/7labsHighmolecularweightandlowabundanceproteinsarelessdetectableby2DE.Proteinswithonlythreeorderofmagnitudecouldbeidentifiedin2DE,butproteinswithfourorderofmagnitudecouldbeidentifiedinshotguntechnology.Morepeptides(redundant)wereidentifiedforhighMWproteinsinshotguntechnology.Somesimilarproteinswereidentified(notproteingroup),suggestingthecriteriafordistinguishingsimilarproteinsshouldbemorestrict.ProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissuesHLPPExperimentalDatabaseMWSDataClient…GUI，WebServices,APIFilesInterchange,DMBSDumpDatasynchronizeDatasubmissionExperimentreports,data,files…

HLPPDataMirrorMWSDataClientMWSDataClientMWSDataClientMWSDataClientMWSServerAdministratorHLPPDataMirrorSubmissionPackagesRepositoryHLPPProjectManagementDatabaseHLPPProjectManagementSystemHLPPParticipatingLaboratoriesHLPPDataCenterTheHLPPDataManagementSystem

ArchitectureBasedonMyWorkSpaceSystem(MWS)CurrentStandarddraftofHLPPSampleInformationProteinextractionConcentrationMeasurementSeparationGel-based2DEGel1D-IEFGel1D-SDSLC-basedRPLCSCXLCSAXLCSECTrapDesaltingDigestionIonsourceMALDIAutoflexUltraflexABI4700ESIQstarQTofMicroLCQLTQMassspectrometryTOFTOFTOFUltraflexABI4700QTOFQstarQTofMicroITMSLCQLTQPeaklistgenerationABI4700QStarAutoflexTurboSequestMasslynxFlexAnalysisPeaklistpreprocessingGPSpeaklistpreprocessProtein/peptideidentificationMascotSequestProteinlistgenerationAutoflexGPSBiotoolsQstarBuildSummaryDTASelectBioworksAutoQuestProteinidentificationisbasedontheanalysisofpeptidesgeneratedbyproteolyicdigestion.Whenthedetectedpeptidesmatchmanyproteinsandcannotidentifyauniqueprotein,theresultwillbepresentedintheformofagroupofpossibleproteins.

DefinitionofProteinGROUP

CriteriaforproteinidentificationIontrapXCorr(highest)≥1.9(1),2.2(2),3.75(3);DeltaCn≥0.1;RSp≤4.ABI-TOF/TOFProteinscore≥59(Rank1forthespot)Q-TOForQ-STARProteinscore>thresholdPeptidescore>“thescoreforidentity”or29MALDI-TOFProteinscore>thresholdTheproteinsinfirstreport(multi-proteinforthemixture)ProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandpreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissuesEvaluationoftheSOPsforthesubcellularfractionationObject:FindtheoptimummethodofpretreatmentSample:LivertissuesofC57miceSubcellular:Mitochondria,Golgi,PM,Nucleus,ER,CytoplasmSamplepretreatmentFreshtissuesFrozenhomogenisedtissuesFrozentissuesPurity(Westernblotting):Freshtissues>frozenhomogenizedtissues>frozentissuesTheyieldofproteins:Freshtissues>frozenhomogenizedtissues>frozentissuesIntegrity(TEM):Freshtissues>frozenhomogenizedtissuesSummaryProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissuesItisagiantambitiousobjectiveandgivesagreatchallengetoembryonicproteomicsItisnecessarytocarryoutapreviewbeforetheformallaunchingofthisprojectScience302:1316,Nov.21,2003Nature425:441,Oct.2,2003AverageThroughputofthePlatform2DESystem

12Gel/3DaysAuto-SpotDigestSystem

1152spots/DayMSandMS/MSSystem

500-1000Spots/DayTheflowchartofCCPITfortheproteinexpressionprofileofhumanfetalliverTheareascalesbeforeandafterisoformprocessingingroupandproteinlevelsrespectively.Inproteinlevel,theredcircularityareasrepresentthenumberofconfirmedproteinsandtheyellowannulusareasrepresentthenumberofpossibleproteins.Non-isogroups19%Extensivelyconfirmed81%Groups41%Confirmed59%Non-isoforms43%Isoforms57%contribute

545

additionalproteins

and

eliminate763

groupscontribute

545

additionalproteins

and

eliminate1335

possible

proteinsIn

Protein

LevelIn

Group

Levelconfirmed:1950possible:

2593extensivelyconfirmed:2495non-isoformpossible:

1258ACBChromosomaldistributionofHFLproteome-encodinggenesFunctionalModulesinplasmamembraneProgressEvaluationoftechnologiesBioinformaticsset-upSamplingandPreparationPreviewwithhumanfetalliverPrimaryanalysisofFrenchlivertissues

ReferenceLabsforHLPPExpressionProfilingProject

FuchuHe&XiaohongQian(BeijingInstituteofRadiationMedicine)RongZeng(ShanghaiInstituteforBiologicalSciences)PengyuanYang(FudanUniversity)SiqiLiu(BeijingGenomicsInstitute,ChineseAcademyofSciences)LauraBeretta(FHCRC,USA)RichardJ.Simpson(RoyalMelbourneHospital,Australia)

AngelikaGorg(TechnischeUniversitatMunchen,Germany)MarkBaker/JunhongSun(APAF,Australia)OutlineBackgroundProgressoftheHLPPInitiationoftheCNHLPPNextworkCNHLPPOverviewThefirstphase:2004-2005Fundingfromcentralgovernment:10millionUSdollars ChineseMOST:jointlyfundedbyNationalProgramon KeyBasicResearchProjects(973),NationalHigh-tech R&DProgram(863),andNationalKeyTechnologiesR&D Programmeininthefirstphase.LocalandInstitutionalfunding:3.5millionUSdollars8subprojects70institutes,universities,andcompaniesinvolvedLong-termsupportasanationalproject(2006~2020)CelebrationofCNHLPPOfficialLaunching(BeijingInstituteofRadiationMedicine)KeylabsBeijingInstituteofRadiationMedicineConsultingcommittee(5)ORFeome&PPISubproject,ZeguangHanChairFuchuHeHeadquartersBPRCModificationProfilesubproject,YunCaiExpressionProfileSubproject,XiaohongQianStructuralBiologySubproject,WeiminGongBioinformaticssubproject,YixueLiNewTechnologysubproject,YukuiZhangAntibodysubproject,Qi-HongSunLocalizationMappingSubproject,XueminZhangMorethan10workshopshavebeenheldBriefprogressofsomesubprojects

ProteinmodificationProteinlocalizationProteinstructureNewproteomicstechnologiesAnalysisofproteinphosphorylationbycombinationofIMAC,PhosphatasewithBiologicalMassSpectrometrym/zPeptidemixtureProteinsCandidatephosphopeptidesIdentificationofphosphorylatedsitesDIGESTIMACMALDI-TOFMSAnalysisPhosphatasedigestMS/MSAnalysisandDatabaseSearchingm/zPhosphopeptideConfirmationm/zMetastableionsofphosphopeptidesO60716GSLApSLDpSLRKP15924GLPSPYNMSpSAPGpSRO75379NLLEDDpSDEEEDFFLRRApSpSKGGGGYTC*QSGSGWDEFTKO95210HSpSWGDVGVGGSLKP16157RDpSRDVDEEKELLDFVPKP02671ADpSGEGDFLAEGGGVRP35221TPEELDDpSDFETEDFDVRP02730

YQpSSPAKPDSSFYKQ13813WRpSLQQLAEERYQSSPAKPDSSFpYKQ15019IYHLPDAEpSDEDEDFKEQTRYQSSPAKPDSpSFYKQ16828SNIpSPNFNFMGQLLDFERRYQSSPAKPDpSSFYKQ99959RLEISPDpSpSPERAHYTHSDYQYSQRP04792GPpSWDPFRLPEEWSQWLGGSSWPGYVRPLPPAAIEpSPAVAAPAYSRQ9H3Z4SLpSTSGESLYHVLGLDKTr:O95810SLEETLHTVDLpSpSDDDLPHDEEALEDpSAEEKVEESRP05386KEEpSEEpSDDDM*GFGLFDKEEpSEEpSDDDMGFGLFDTr:Q8NEN5pSQpSLPTTLLSPVRP08559YHGHpSMSDPGVSYRTr:Q9HAP4ESEALPEKEGEELGEGERPEEDAAALELpSpSDEAVEVEEVIEESRP11277TpSPVSLWSRLpSpSSWESLQPEPSHPYTr:Q9NQA7WLDEpSDAEMELRIdentifiedphosphopeptidesequencesfromHFLProteinisolationandpurificationbylectin(Glycosylation)Co-localizationandlocomotionoftheGFPfusionproteinandDsRedDsRed-MemGreenmergeGFPGFP-fusedPrTNF-

treatedGFP-fusedmutantTNF-

treatedGFP-fusedPrStructuralproteomicsCloned900genesfromliverExpressedproductsofmorethan100genesDeterminedthestructuresof3proteinsbyX-rayDeterminedthestructuresof2proteinsbyNMRDiphosphoglyceratemutase(DPGM)X-rayProgrammedcelldeath5(PDCD5)NMRBeijing(National)ProteomeResearchCenterChineseProteomeDevelopmentCenterTheAntibodyBankofHumanLiverProteomeProjectEstablishment,characterization,andapplicationofantibodiesagainsthumanliverproteins

Qi-HongSunBeijingInstituteofRadiationMedicine(BIRM)BeijingProteomeResearchCenter(BPRC)AntigensMyelomacellsHybridomacellsmAbsDiagnosticsTherapeuticsProteomicsResearch

ProfilingIdentificationQuantificationLocalizationModificationInteractionFunctionLargeantibodycollectionforproteomicsAntibodyDatabaseAntibodyBankSamples5000liverproteinsHLPPAntibodyBankAntibodychips10%ofHLPPAntibodyBankStandardsanddatabaseofHUPOHLPP/HAIMicroarrayandotherantibody-basedtechnology90%10%HLPPAntibodyBankatPilotPhaseAntigenpreparation

Fractionatedliverproteins-Unknown/native/multi-antigensRecombinantproteinsSynthesizedpeptidesAntibodygenerationHybridomacelllines–mAbs(murine/rabbit)Polyclonalantibodies–IgG(Rabbit)/orIgY(chicken)AntibodycharacterizationandapplicationClas