新填料應(yīng)用Capto系列及其

內(nèi)Capto系列填料及其應(yīng)多模式新型填料及其應(yīng)新型親和填料及其應(yīng)新型凝膠過濾預(yù)裝Capto系列填料及其應(yīng)CaptoCapto系列CaptoCapto下游處 表達(dá)量越來越高，由以前的mg/l到現(xiàn)在的Capto什么是CaptoTM是一 商CaptoTMCaptoTMCaptoTMCaptoTM瓊脂糖基架的優(yōu)安全性 部門所認(rèn)Capto高流速瓊脂Chemicalmodificationof

Sepharose?4FastO2OOOHO2OOOH OO

(High

Sepharose6FastFlow(Mediumporosity)(Highporosity)

CaptoFamily(Mediumporosity)ROOOROOOOOOOOR靈活的操作窗“Increasing“IncreasingtheWindowCapto結(jié)構(gòu)特通過提高內(nèi)表面積提高載

改善質(zhì)量傳ConventionalPorous GraftedLayerofaHydrophilicCapto離子交換填CaptoQ,CaptoSandCapto

3Capto?S3BaseHighflowagarose90μm(average)

SurfaceAnimal-free

CaptoQQuaternaryNH+NOORRNNORCaptoDEAE高動(dòng)態(tài)載量更寬的工作范圍，更小占地面BedheightFlowvelocityina1mdiametercolumn(cm/h)Sepharose6FastMeasuredflowvelocityforCaptoQat3barwith20cmbedheightinFineLINE?100:850BedheightFlowvelocityina1mdiameterSepharose6FastMeasuredflowvelocityforCaptoQat3barwith45cmbedheightinFineLINE100:600Capto?產(chǎn)品的優(yōu)ReducedPlateHeight(h)1123Columnefficiencydataforthreecolumnpackingsat20cmbedCapto填料 支完善 支持文件Capto系列陰離子填料—CaptoCaptoDEAE具有非常明顯的弱離子交換的屬性HNN+OORR++NORpH~5:絕大部分的基團(tuán)質(zhì)子化后代帶有正電增加pH:團(tuán)逐漸去質(zhì)子pH1011有的弱陰離子交換基團(tuán)失去質(zhì)子不帶電荷，只剩下CaptoDEAE與CaptoQ洗脫條Capto?Capto?Capto5–0.25ml/min0-40%BunderCaptoQ與CaptoDEAE如何選擇一般情況下在開始階段推薦使用強(qiáng)離子交換填(CaptoQ，CaptoDEAE.由于CaptoDEAE在高pH下有去質(zhì)子化的作用，減弱與蛋白的結(jié)CaptoQ/SPImpRes--快流速高分辨率40μmCaptoHiScreenQHiScreenHiScreenQHiScreenCaptoQHiScreenQCapto系列陽離子填CaptoS90μm120mgHighbindingcapacityMediumresolution

CaptoSPImpRes40μm70mg~50mgMAb/mLMediumbindingHigh

CaptoS5090mg100mgMAb/mLHighbindingcapacityHighresolutionLaunched

Launched

LaunchedSep.CaptoSImpAct是目前具有最高載量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--疏水性強(qiáng)弱序列 疏水 不同流速下CaptoPhenyl的BSA載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應(yīng)用舉

Presentedat6thHICconferenceMarchAerovanceCo.Productrelatedimpurities(mainlysolubleaggregates)Capto?Butyl的優(yōu)良表ProducutionbyLinearProducutionbyStep

FlowTimeRecoveryPurityMaterialwasappliedtoa0.5x14.5cmTricornTMcolumnpackedwithCapto?Butylwithloadingdensityof12g/L,attheindicatedflowvelocities.ProductrecoveryandpurityweredeterminedbyRP-HPLC.73.20.067<1.251.16x106<11Determinedusing1.6x28cmCaptoButylwithloaddensityof12gprotein/L2BelowdetectionlimitofAerovance公司對CaptoButyl的評流速和柱床的提高大大增加了生產(chǎn)能卓越的純化性容易裝CaptoMMC—Captoadhere—有效去除聚體、HCP及雜CaptoCore700—純 及大分CaptoMMCMMC-+75μm

MultimodalweakcationMultipletypesofinteractionsionicinteractionhydrophobicinteractionhydrogenbondingCaptoMMC耐受高鹽上CaptoMMCFab,Ip8.6,Mw~501Mphosphatebuffer,pH6,1MNaCl

1MNaCl0.15M 快速捕快速捕獲大體積高電導(dǎo)樣品，減少工藝Capto+OONO O+OONO+OHBaseHighflowagarose75μm(average)

adhere-Multimodalstrong

Multipletypesofinteractionsionicinteractionhydrophobicinteractionhydrogenbonding有效去 體，殘有效去 體，殘 1782323629475應(yīng)用于流腦多糖新工PhenolHigh-speed

PSNucleicAcid

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

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

CrudePSdissolveandSDCaddedCaptoDEAE&AdhereFTmode7MeningococcusVaccineDSPCaptoCore天然瓊脂糖基架外激活內(nèi)核-帶辛氨基(多通道配體-目標(biāo)物-進(jìn)入孔內(nèi)的小分子CaptoCore700優(yōu)點(diǎn)

*comparedtogelCaptoCore700vsGelinHCPinHCPandDNAintotalinHCPandDNAinRecovery(HA*)[%]HostCellProteinRemoval[%]AmountofloadedfeedmaterialDilutionSepharose?4CaptoCore1SameHCPremoval recoveryasgel100timeshigherloadthangelfiltrationincreasesproductivityNodilution*HA=Hemagglutinin,mainsurfaceproteinon usedin基于雞胚的流基于雞胚的流 純OvalbuminreductionCaptoCore可以使用CaptoCore700純化的Allmarketedhumanviralvaccinesarepossible AlllargemoleculesthatdonotentertheCapto?Core700beadscanbeCaptoCore700IgM純 質(zhì)粒 ...其內(nèi)毒素的去酚紅的去樣品準(zhǔn)用 治療 載膜蛋白純 純>95%HCPremovalat75%

RemovalofRNA,HCP,endotoxinsfromplasmid

批量吸附應(yīng)用于抗體純化的新填料及技MabSelectSuRe用于抗體片段純化的新填料及技ProteinAMabSelectSuReMabSelectrProtASepharose

MabSelectXtranProtASepharose

rmProtASepharose

MabSelectSuReLX MabSelectSuReLX動(dòng)態(tài)載量MabSelectSuRe提高h(yuǎn)uman

monoclonalMabSelectSuReLXchromatographyMabSelectSuReLXchromatographymediumconsistentlyprovideshigherbindingcapacityversusMabSelectSuRe.NaOH進(jìn)行CIP，清潔更徹403CIP3000.1MNaOH400.1MNaOH403無需額外處理/QC0.5MNaOH有效滅活，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耐堿SuRe配基由5IgGBinding

cellwallbindingEDABCXEDEDABCXEDABCXMr~42rProteinMr34(Aspreplacedwithalkali-stableaminoacidN

SuReLigandforMabselectMrOO +HS–ProtOO

S–OOOOSingle-pointattachedthioetherMabSelectSuReLX耐堿性更強(qiáng)，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更好的壓力流速性5抗體和抗體片全

Antibody Chain mended AlternativeKappaLambda CaptoKappaLambda Kappa

CaptoKappaVHKappaVH3heavy CaptoL—對廣泛的抗體片段都具有較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用于血漿蛋新填應(yīng)Alpha-1AntitrypsinAlpha-1FactorFactorCaptoNiSepharoseTALON?Superflow?--LMW-SDSMarker 6.Start3. 8.3. 8.螯合螯合結(jié)合、、洗脫時(shí)4. 9.4. 9.提高His蛋白純純化真核細(xì)胞胞外分泌表達(dá)蛋白的因?yàn)榧兓盍虾图?xì)胞培養(yǎng)基之間的不相容性所目標(biāo)蛋白不結(jié)合或者結(jié)合載量很低目標(biāo)樣品體積很大（>5L)現(xiàn)有預(yù)處理的方法-使用過濾置換緩沖-濃預(yù)預(yù)處理對于敏感蛋白會造成時(shí)間上的浪費(fèi)以及潛在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,同樣適用于使傳統(tǒng)IMAC填料金屬離子脫落的樣