新填料应用Capto系列及其

内Capto系列填料及其应多模式新型填料及其应新型亲和填料及其应新型凝胶过滤预装Capto系列填料及其应CaptoCapto系列CaptoCapto下游处 表达量越来越高，由以前的mg/l到现在的Capto什么是CaptoTM是一 商CaptoTMCaptoTMCaptoTMCaptoTM琼脂糖基架的优安全性 部门所认Capto高流速琼脂Chemicalmodificationof

Sepharose™4FastO2OOOHO2OOOH OO

(High

Sepharose6FastFlow(Mediumporosity)(Highporosity)

CaptoFamily(Mediumporosity)ROOOROOOOOOOOR灵活的操作窗“Increasing“IncreasingtheWindowCapto结构特通过提高内表面积提高载

改善质量传ConventionalPorous GraftedLayerofaHydrophilicCapto离子交换填CaptoQ,CaptoSandCapto

3Capto™S3BaseHighflowagarose90µm(average)

SurfaceAnimal-free

CaptoQQuaternaryNH+NOORRNNORCaptoDEAE高动态载量更宽的工作范围，更小占地面BedheightFlowvelocityina1mdiametercolumn(cm/h)Sepharose6FastMeasuredflowvelocityforCaptoQat3barwith20cmbedheightinFineLINE™100:850BedheightFlowvelocityina1mdiameterSepharose6FastMeasuredflowvelocityforCaptoQat3barwith45cmbedheightinFineLINE100:600Capto™产品的优ReducedPlateHeight(h)1123Columnefficiencydataforthreecolumnpackingsat20cmbedCapto填料 支完善 支持文件Capto系列阴离子填料—CaptoCaptoDEAE具有非常明显的弱离子交换的属性HNN+OORR++NORpH~5:绝大部分的基团质子化后代带有正电增加pH:团逐渐去质子pH1011有的弱阴离子交换基团失去质子不带电荷，只剩下CaptoDEAE与CaptoQ洗脱条Capto™Capto™Capto5–0.25ml/min0-40%BunderCaptoQ与CaptoDEAE如何选择一般情况下在开始阶段推荐使用强离子交换填(CaptoQ，CaptoDEAE.由于CaptoDEAE在高pH下有去质子化的作用，减弱与蛋白的结CaptoQ/SPImpRes--快流速高分辨率40μmCaptoHiScreenQHiScreenHiScreenQHiScreenCaptoQHiScreenQCapto系列阳离子填CaptoS90µm120mgHighbindingcapacityMediumresolution

CaptoSPImpRes40µm70mg~50mgMAb/mLMediumbindingHigh

CaptoS5090mg100mgMAb/mLHighbindingcapacityHighresolutionLaunched

Launched

LaunchedSep.CaptoSImpAct是目前具有最高载量DBCatdifferentresidence

DBCfordifferentMAbsandDBCDBC10%breakthrough

mAbA,pH5.0mAbA,pH6.0mAb CaptoSPEshmuno™Fractogel™Capto™SColumn:Tricorn™Sample:MAbB(9.4mg/mL)instartbufferStartbuffer:50mMsodiumacetate,pH5.5Samplesrunintriplicate

SO3- CaptoSImpAct具有更好的分辨Startbuffer: 50mMsodiumacetate,pH5.3(3mS)Elutionbuffer: 0to350mMNaCl,lineargradient,20CVResidencetime: 5.4minCapto™SImpAct(64 CaptoSPImpRes(34Aggregate

Aggregate

HCP

0

Capto系列疏水填料--疏水性强弱序列 疏水 不同流速下CaptoPhenyl的BSA载86420

150cm/h<=>4,0min.residencetime(backpressure~3bar)300cm/h<=>2,0min.residencetime(backpressure~3bar)900cm/h<=>0,7min.residencetime(backpressure~5bar)1200cm/h<=>0,5min.residencetime(backpressure FlowrateLoadingbuffer1.2M(NH4)2SO4+100mMNaH2PO4,elutionbuffer100mMNaH2PO4with10%CaptoButyl应用举

Presentedat6thHICconferenceMarchAerovanceCo.Productrelatedimpurities(mainlysolubleaggregates)Capto™Butyl的优良表ProducutionbyLinearProducutionbyStep

FlowTimeRecoveryPurityMaterialwasappliedtoa0.5x14.5cmTricornTMcolumnpackedwithCapto™Butylwithloadingdensityof12g/L,attheindicatedflowvelocities.ProductrecoveryandpurityweredeterminedbyRP-HPLC.73.20.067<1.251.16x106<11Determinedusing1.6x28cmCaptoButylwithloaddensityof12gprotein/L2BelowdetectionlimitofAerovance公司对CaptoButyl的评流速和柱床的提高大大增加了生产能卓越的纯化性容易装CaptoMMC—Captoadhere—有效去除聚体、HCP及杂CaptoCore700—纯 及大分CaptoMMCMMC-+75µm

MultimodalweakcationMultipletypesofinteractionsionicinteractionhydrophobicinteractionhydrogenbondingCaptoMMC耐受高盐上CaptoMMCFab,Ip8.6,Mw~501Mphosphatebuffer,pH6,1MNaCl

1MNaCl0.15M 快速捕快速捕获大体积高电导样品，减少工艺Capto+OONO O+OONO+OHBaseHighflowagarose75µm(average)

adhere-Multimodalstrong

Multipletypesofinteractionsionicinteractionhydrophobicinteractionhydrogenbonding有效去 体，残有效去 体，残 1782323629475应用于流脑多糖新工PhenolHigh-speed

PSNucleicAcid

Cultureinbioreactorfor6-8hoursHarvestandinactivationusingmethanolAddcetavlontoprecipitatePSAddCaCl2Add95%ethanolto25%Add95%ethanolto80%concentrationPrecipitateanddryPSProtein

Add1:10SaturatedsodiumacetateAddcoldphenol(threetimes)AddCaCl2,CentrifugebyAdd95%ethanolto80%concentrationWashprecipitationwithethanolandacetone,twotimeseach

CrudePSdissolveandSDCaddedCaptoDEAE&AdhereFTmode7MeningococcusVaccineDSPCaptoCore天然琼脂糖基架外激活内核-带辛氨基(多通道配体-目标物-进入孔内的小分子CaptoCore700优点

*comparedtogelCaptoCore700vsGelinHCPinHCPandDNAintotalinHCPandDNAinRecovery(HA*)[%]HostCellProteinRemoval[%]AmountofloadedfeedmaterialDilutionSepharose™4CaptoCore1SameHCPremoval recoveryasgel100timeshigherloadthangelfiltrationincreasesproductivityNodilution*HA=Hemagglutinin,mainsurfaceproteinon usedin基于鸡胚的流基于鸡胚的流 纯OvalbuminreductionCaptoCore可以使用CaptoCore700纯化的Allmarketedhumanviralvaccinesarepossible AlllargemoleculesthatdonotentertheCapto™Core700beadscanbeCaptoCore700IgM纯 质粒 ...其内毒素的去酚红的去样品准用 治疗 载膜蛋白纯 纯>95%HCPremovalat75%

RemovalofRNA,HCP,endotoxinsfromplasmid

批量吸附应用于抗体纯化的新填料及技MabSelectSuRe用于抗体片段纯化的新填料及技ProteinAMabSelectSuReMabSelectrProtASepharose

MabSelectXtranProtASepharose

rmProtASepharose

MabSelectSuReLX MabSelectSuReLX动态载量MabSelectSuRe提高human

monoclonalMabSelectSuReLXchromatographyMabSelectSuReLXchromatographymediumconsistentlyprovideshigherbindingcapacityversusMabSelectSuRe.NaOH进行CIP，清洁更彻403CIP3000.1MNaOH400.1MNaOH403无需额外处理/QC0.5MNaOH有效灭活，降低生物Increasedviral

IncreasedbioburdenC.1A.1bacteriagram1S.bacteriagram1 0.1 0.5 age,0.5MNaOHresults0.5LRVhigherinactivation0.1M

1forreductiontobelowdetectionlimitof<3organisms0.5MNaOHeffectivelyinactivatesbacteria,moldandyeast0.1M0.50.1M0.5M0.5MNaOHrapidlyinactivatesendotoxin(seecharttoright)CIP=cleaninginplace,LRV=logreductionMabSelectSuRe™LX|May Approvedforexternal Version耐碱SuRe配基由5IgGBinding

cellwallbindingEDABCXEDEDABCXEDABCXMr~42rProteinMr34(Aspreplacedwithalkali-stableaminoacidN

SuReLigandforMabselectMrOO +HS–ProtOO

S–OOOOSingle-pointattachedthioetherMabSelectSuReLX耐碱性更强，DBCresultsobtainedwith0.5MNaOHattworesidencetimes(RT6minor2.4min)forMabSelectSuRecomparedwithMabSelectSuReLXatRT6min.

>95%ofDBCretainedafter300cycleswith0.1MNaOH>80%ofDBCretainedafter150cycleswith0.5MNaOHMabSelectSuReLXchromatographymediumexhibits20%longerusefullifetimecomparedwithMabSelectSuRemediumMabSelectSuReLX|May ApprovedforexternalVersion MabSelectSuReLX具有和MabSelectHostHostcellprotein(HCP) HCPclearance0 CycleMabSelectSuRe MabSelectLeachedproteinA ProteinALeachedproteinA50Cycle

NOTE:Feedwaschangedafter87MabSelectSuRe MabSelectMabSelectSuReLX|May2014 ApprovedforexternaluseVersionAA更好的压力流速性5抗体和抗体片全

Antibody Chain mended AlternativeKappaLambda CaptoKappaLambda Kappa

CaptoKappaVHKappaVH3heavy CaptoL—对广泛的抗体片段都具有较DynamicBreakthroughCapacity(Qb10mg/ml)

DynamicBreakthroughCapacity(mg/mlandmolareqv.)**FabkindlyprovidedbyUCB

**Customer

0.5µmol/ml0.40.5µmol/ml0.4scFv/ml1.4Dab/ml1.3Dab/ml6E.coliHCPClearance3.6log(evaluatedby93.2%6.5%UVAbsorbanceUVAbsorbance acetateat3210pH

pH2.5Load:ClarifiedE.colifermentationbrothatpH7.0

Load:11.4mg/mlresinResidencetime:4minAlpha-1AntitrypsinCapto用于血浆蛋新填应Alpha-1AntitrypsinAlpha-1FactorFactorCaptoNiSepharoseTALON®Superflow™--LMW-SDSMarker 6.Start3. 8.3. 8.螯合螯合结合、、洗脱时4. 9.4. 9.提高His蛋白纯纯化真核细胞胞外分泌表达蛋白的因为纯化填料和细胞培养基之间的不相容性所目标蛋白不结合或者结合载量很低目标样品体积很大（>5L)现有预处理的方法-使用过滤置换缓冲-浓预预处理对于敏感蛋白会造成时间上的浪费以及潜在NiSepharoseexcelNEWNEWCellRemovalofFinalandIMACconventionalNiCellRemovalofNiSepharose™HisMagSepharoseTimesavingsupto50%NiSepharose™excelvs.NiSepharose6Sample:250mlmPAI-1-(his)8secretedintoGIBCO™CDmedium,pHBlue–NiSepharoseexcelpackedinTricorn™5/50:Purity>98%Green-NiSepharose6FFpackedinTricorn5/50:Norecoveryof

LMW-SDSMarkerStartNiSepharoseexcel,NiSepharoseexcel,NiSepharoseexcel,NiSepharose6FF,NiSepharose6FF,NiSepharose6FF,同样适用于使传统IMAC填料金属离子脱落的样