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血液制品層析工藝中TOYOPEARL填料東曹(TOSOH)企業(yè)生命科學(xué)部門PlasmaProductsReviewApplicationonPlasmaProteinContent2ProteinConcentrationinPlasmaIndicationAlbumin40g/LVolumerestorationaftertrauma,shock,burnsAlpha1ProteinaseInhibitor1.5mg/mLHereditaryemphysemaAnti-DIgGTitervariesaRhprophylaxisinpregnancyandchildbirthAntithrombinIII100ug/mLAntithrombinIIIdeficiencyC1-inhibitor170ug/mLHereditaryangioedemaFactorIX10ug/mLHemophiliaBFactorVIII0.5ug/mLFactorVIIdeficiencyFactorXI0.3ug/mLHemophiliaBFactorXIII30ug/mLFactorXIIIdeficiencyFibrinogen3g/LTissuesealantcomponentFibronectin300ug/mLWoundhealingHepatitisBIgGTitervariesaHepatitisimmunityImmunoglobulinGUpto12.5g/LPrimaryandsecondaryimmunedeficiencyMeaslesIgGTitervariesaMeaslesprotectionandtreatmentProteinC4ug/mLNeonatalthrombosisRabiesIgGTitervariesaRabiesriskTetanusIgGTitervariesaTetanusprotectionandtreatmentThrombin150ug/mLbTissuesealantcomponentVaricellaZosterIgGTitervariesaChickenpoxprotectionVonWillebrandFactor10ugVonWillebrand’sdiseaseaTiters(antibodyconcentrations)varyinug/mLranges.bAsprothrombinPlasmaProteinsofCurrentInterest3PlasmaProductsinChina4PlasmaprecipitationsupernatantCryoprecipitateFactorVIIIFractionICentrifugation4oCEthanoltreatment,CentrifugationsupernatantprecipitationFibrinogenFactorIXAbtithrombinIIICohnColdEthanolFractionationProcess(Continued)5precipitationsupernatantFractionIVFractionVEthanoltreatment,CentrifugationsupernatantprecipitationHaptoglobinAbtithrombinIIIAlbuminFractionII+IIIsupernatantprecipitationprecipitationEthanoltreatment,CentrifugationFractionIIGlobulin(forintramuscular)Globulin(forintravenous)EthanoltreatmentEthanoltreatment,CentrifugationCohnColdEthanolFractionationProcess6ProductionCapacityofPlasmaProteinsCapacity;MillionL/YCompanieswithyellowbarhaveappliedToyopearltotheirprocess.7CSLBioplasma,AustraliaCommonwealthSerumLaboratory(CSL)InternationalpharmaceuticalcompanybasedinAustraliaasCSLgroupsAllprocesseswereapplied

bychromatographyforCohnethanolfractionationexceptFraction1in1994Thefourthlargestsupplierofplasmaproteins(2023)GlobalcompanywithmanufacturingplantinUSandEuropeaswell.ConsultanttoChineseplasmaproteinsuppliers8WhyCSLBioplasmaAppliesChromatography?

CSLBioplasmadescribeschromatographyas;HighlycosteffectiveprocessHighproductionefficiency(IVIG;5.5g/L)Highquality(Albumin;99%w/v)Achievableofautomatic,closed-processEasyextractionandseparationofby-products(i.e.otherplasmaproteins)Advancedtechniqueinbio-industry9ProductionProcessforPlasmaProteinsSizeExclusionChromatographyHW-55FIon-ExchangeChromatographyDEAE-650M,QAE-550C,SuperQ-650S,MSP-650M,SP-550CHydrophobicInteractionChromatographyButyl-650M,C,Phenyl-650S,MAffinityChromatographyAF-Formyl650M,AF-HeparinHC650M10ApplicationofPlasmaProducts11PurificationofAlbumin12PurificationofAlbuminbyAIECConditionColumn:ToyoScreenSuperQ-650M(1mL)Eluent:(A)50mMTris-HCl(B)50mMTris-HCl+0.5MNaClGradient:(A)->(B)Linear30minFlowrate:1mL/minDetection:UV(280nm)Sample:HumanAlbumin,Globulin1g/LInjection:100uL13PurificationofAlbuminbyCIECConditionColumn:ToyoScreenSP-650M(1mL)Eluent:(A)50mMAceticbuffer(pH5.0)(B)50mMAceticbuffer+0.5MNaCl(pH5.0)Gradient:(A)->(B)Linear30minFlowrate:1mL/minSample:HumanAlbumin,Globulin1g/LInjection:100uL14AnionexchangerCationexchangerAlbuminGlobulinFractionVLoadingElutionLoadingElutionConsiderationofAlbuminPurification15Chromatographyinplasmafractionation:benefitandfuturetrendsT.Burnouf,J.Chromatgr.B664(1995)3-15.“TOYOPEARLDEAE-650MisusedtoproduceahighlypurifiedFVIIIconcentrate,......”PurificationofFactorVIII16StartingfromCohnfractionation,chromatographyonTOYOPEARL?DEAE-650MresultsinahighlypurifiedclottingfactorfortreatmentofhemophiliaA.MethodislicensedfromOctapharmaandincludestwovirusinactivationsteps:

solvent/detergentandheating.Scaleishundredsofmillionunitsperyear.TOYOPEARL?DEAE-650Mchromatographyistheresinofchoice.PurificationofFactorVIII(Octanate?)1718ConditionColumnsize:6.0mmID*4cmL(1.13ml)Mobilephase:Loading:0.4mg/mlAT-IIIin0.01MTris-HClcontaining0.15MNaClpH7.5LinearVelocity:300cm/minDetection:UV(280nm)ResinAT-IIIbindingcapacity(10%leakage,mg/ml-gel)NewHeparin-650M 3.9HeparinSepharose6FF 2.4Heparin-650M 1.9Figure1Comparisonofbreakthroughcurveat300cm/hComparisonofdynamicAT-IIIbindingcapacityat300cm/hPurificationofAT-III19Conditioncolumnsize:6.0mmID*4cmL(1.13ml)Mobilephase:Loading:0.4mg/mlAT-IIIin0.01MTris-HClcontaining0.15MNaClpH7.5Detection:UV(280nm)ComparisonofdynamicbindingcapacityPurificationofAT-III20PeakAPeakBbufferAbufferBAT-III

PurificationofAT-IIIfromhumanplasmaonHeparin-Toyopearl650M

Columnsize:6.0mmI.D.X4.0cmSample:Humanplasma18mlBuffers:Adsorption,0.02MCitrate+0.12MNaCl

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