产品资料

Polyplus Transfection/FectoPRO^®/Polyplus Transfection-161253/1 Ea

Amazingproteinandantibodyproductionyields
Cost-effectiveTransientGeneExpressionusinglowerplasmidDNAamount
CompatIBLewithvariousmammalianexpressionmedia&cellssystems
Compliantwithbiomanufacturingguidelines

Specifications:

Reagent	FectoPRO^®
Moleculedelivered	DNA
Applications	Bioproduction
Celltypes	SUSPensionCHO,HEK-293,andderivatives
Numberoftransfections	1mLofFectoPRO^®issufficienttotransfect800mltomorethan1literofcellculture
Storage	4°C,stablefor1to2yearswhenstoredappropriately
Providedwith	FectoPRO^®Booster

Summary:

Forthefirsttime,achievesuperiorproteinyieldsusingtransienttransfectionandgeneexpression.

Orderinginformation:

|||116-001##1ml##1ml##|||116-010##10ml##10ml##|||116-040##4x10ml##4x10ml##|||704-10##-##10ml##|||

Description:

Amazingproteinyields

FectoPRO^®enablesamazinglyhighproteinandantibodyyieldsinbothCHOandHEK-293cellsgrowninsuspensioninserum-freemedia(Fig1).

FectoPRO - antibody yields CHO cells — **Fig.1.FectoPRO^®givesamazingantibodyproductionyieldsinsuspensionCHOcells.**Cellswereseededat1x106cells/mlin30mlofFreeStyle™CHOExpressionMediumandtransfectedwithFectoPRO^®(0.8µgDNA/ml)orFreeStyle™MAXReagent(1.25µgDNA/ml)followingrecommendedprotocolsfortherespectivereagents.RecombinantmouseIgGproductionyieldwasassayed1to6daysaftertransfectionusingproteinGbiosensor(fortéBIO^®octetRED96system).FreeStyle™isatrademarkofLifeTechnologies™Corporation.DatakindlyprovidedbyProteoGenixSAS(France).

FectoPRO^®largelyoutperformsothertransfectionreagentssuchaslinearPEIcommonlyusedinBioproductionprocessesintermsoftransfectionefficiency(Fig.2),leADIngtosuperiorproteinproductionyields(Fig.3).

FectoPRO - Microscopie — **Fig.2.FectoPRO^®giveshightransfectionefficiencyinsuspensionCHOcells**.FreeStyle™CHO-Scellswereseededat1x10⁶cells/mlin30mlofFreeStyle™CHOExpressionMediumandtransfectedwithFectoPRO^®+FectoPRO^®BoosterorFreeStyle™MAXReagentusing0.4µg/mland1.25µg/mlofaGFPexpressingplasmid,respectively.GFPexpressionwasassayed24haftertransfectionusingfluorescencemicroscopy.

FectoPRO - CHO cells

FectoPRO - HEK-293F cells — **Fig.3.FectoPRO^®givessuperiorproteinproductionyieldsinsuspensionCHOandHEK-293cells**.FreeStyle™CHO-SandHEK-293Fcellswereseededat1x10⁶cells/mlin30mlofFreeStyle™CHOor293ExpressionMediumandtransfectedwithFectoPRO^®+FectoPRO^®Booster(0.4µgDNA/ml),FreeStyle™MAXReagent(1.25µgDNA/ml)orPEIpro^®(1µgDNA/ml)followingrecommendedprotocolsfortherespectivereagents.IgG3-Fc(34kDa)productionwasassayed72h(CHOcells)and120h(HEK-293cells)aftertransfectionbyproteinGaffinityquantification(HPLC)andWesternBlot.StandardcorrespondstopurifiedIgG3-Fc(34kDa).

Cost-effective:useslowplasmidDNAamounts

HighqualityplasmidDNApreparationcanbeverycostly,especiallywhenlargeDNAamountsandmanyplasmidsarerequiredforlarge-scaleproteinproduction.FectoPRO^®saves40to60%onDNAcostbyusing0.4to0.6µgDNA/106cells/mlofcellculture;hencemakingthiskitanaffordableandeconomicalsolutionforBioproductionprocesses.

FectoPRO - Table comparison competitor — **Table1.AmountsofDNAandvolumesofreagent(FectoPRO^®andcompetitor)fortransfectionaccordingtomanufacturers’recommendations.**

Sustainedproteinproduction

Thedurationofproteinproductionprocesseshasincreasedinthepastfewyears,andmostoftheproteinproductionprocessesnowlast1-2weekstoobtainoptimalamountofproducedprotein.FectoPRO^®hasbeendevelopedtoensuresustainedproteinproductionoverseveraldays,whilemaintaininghighcellviABIlityforlongerlastingproteinproduction(Fig.4.).

FectoPRO - Production + viability in HEK-293 — **Fig.4.FectoPRO^®givessustainedproteinproductioninsuspensioncells**.FreeStyle™HEK-293Fcellswereseededat1x106cells/mlin30mlofFreeStyle™293ExpressionMediumandtransfectedwithFectoPRO^®+FectoPRO^®Booster(0.5µgDNA/ml)followingthestandardprotocol.IgG3-FcproductionwasassayedafterdifferenttimepointsbyproteinGaffinityquantification(HPLC).SimilarresultsareobtainedwithFreeStyle™CHO-S(datanotshown).

Easilyscalable

Streamlineyourbioproductionprocesswitharobustprotocolthatiseasilyadaptedtovariousculturevessels.FectoPRO^®-mediatedtransfectioncanbescaledlinearlyfrom30mluptolitersofcellculturewithoutprotocoladaptation(Fig.5.).

FectoPRO - CHO viability FectoPRO - CHO protein production

FectoPRO - Scalability — **Fig.5.FectoPRO^®iseasilyscalableandreproducibleinCHOcells**.FreeStyle™CHO-Scellswereseededat1x10⁶cells/mlin30ml,100ml,300mlor1LofFreeStyle™CHOExpressionMediumandtransfectedwithFectoPRO^®+FectoPRO^®Booster(0.5µgDNA/ml).Atday1to6,cellviabilitywasassayedusingTrypanblueexclusionassay(A)andIgG3-FcproductionwasassayedbyproteinGaffinityquantification(HPLC)(B)andWesternBlot(C).Standard(C)correspondstopurifiedIgG3-Fc(34kDa).

Reproducibleproductionyields

FectoPRO^®ismanufacturedandformulatedusingahighlycontrolledproductionprocess.Thisensureshighlyreproducibleresultsfromexperimenttoexperiment.Moreover,inordertomeetcurrentregulatoryguidelines,FectoPRO^®isreleasedusingadvancedqualitycontrolsincludingaspecificationforproteinproductivitythatguaranteesexcellentlot-to-lotconsistency(Fig.6).

FectoPRO - Reproductibility batch to batch — **Fig.6.FectoPRO^®givesexcellentlot-to-lotreproducibility**.SuspensionCHOcellswereseededat1x10⁶cells/mlin30mlofCHOmediumandtransfectedusing3differentlotsofFectoPRO^®(CCIII078,CCIII104orCCIII106)followingthestandardprotocol.72haftertransfection,IgG3-FcproductionwasassayedbyproteinGaffinityquantification.SimilarresultsareobtainedonsuspensionHEK-293cells(datanotshown).

Compatiblewithmammalianexpressionsystems

Unlikeothercommerciallyavailablereagents,FectoPRO^®transfectionkitissuitableforbothCHOandHEK-293cells(Fig.2and3).Thereagentiscompatiblewithvariousmediaformulations;hencelabimplementationiseasyandstraightforward(Fig.7,Table2).

FectoPRO - protein production — **Fig.7.FectoPRO^®givessuperiorproteinproductionyieldsinsuspensionCHOcellsgrownindifferentmedia**.CHOcellswereseededat1x10⁶cells/mlin30mlofFreeStyle™CHOExpressionMedium,CDCHOMediumorPro-CHO™4MediumandtransfectedwithFectoPRO^®+FectoPRO^®BoosterorFreeStyle™MAXReagentfollowingrecommendedprotocolsfortherespectivereagents.IgG3-Fcproductionwasassayed72haftertransfectionbyproteinGaffinityquantification(HPLC).

FectoPRO - Table media CHO — **Table2.ProteinyieldsusingFectoPRO^®indifferentculturemediaforCHOcells.**

FectoPRO - Table media HEK — **Table3.ProteinyieldsusingFectoPRO^®indifferentculturemediaforHEK-293cells.**

FreeStyle^TM,CDFortiCHO^TM,CDOptiCHO^TM,Expi293^TMandCD293aretrademarksofGibco^TM.
Pro-CHO^TM,PowerCHO^TMandPro293^TMaretrademarksofLonza^TMGroup.
HyClone^TM,HyCell^TMandTransFx^TMaretrademarksofGEHealthcare^TM.
BalanCD^®andTransfectory^TMaretrademarksofIrvineScientific^TM.

FectoPROisalsoperfectlysuitedforvirusproduction.

Applications:

Transientproteinproduction

Proteinproductionconsistsinoverexpressingarecombinantproteinorpeptideinacelllineatsmall/mediumtolargescale.Thebiomoleculesproducedcanbeusedforresearchpurposeslikestructuralcharacterizationallthewaydowntotherapeuticormedicalapplications.

AvailableapplicationNotes:

OptimizationofGlycoproteinExpressionbyTransientTransfectioninHEK293F/Ssuspensioncells

SuperiorproteinyieldsinsuspensionCHOcellsusingFectoPRO-mediatedtransienttransfectioninCELLSTAR^®CELLreactor™

ImprovedProteinyieldsinCHOcellsusinganoveltransfectionreagentforTGE:FectoPRO

Readmore…

HighdensitycellsystemsforproteinproductionusingFectoPRO

RecombinantproteinproductionfortherapyanddiagnosticapplicationsismainlyachievedusingHEK-293andCHOcellscultivatedinsuspensioncultureswithoutserum.Thelatterhasbeenremovedfromrawmaterialsforvariousreasons(costs,downstreampurificationsteps,reproducibility…)whilecultureinsuspensionhasbeenimplementedtoincreaseproductivity.Improvementsincultureprocessesandcelllineengineeringledtothegenerationofhigh-densitymammaliantransientproteinexpressionsystems.

Readmore…

Citations:

Batonick,M.,Kiss,M.M.,Fuller,E.P.,Magadan,C.M., Holland,E.G.,Zhao,Q.,Wang,D.,Kay,B.K.,Weiner,M.P.(2016). pMINERVA:Adonor-acceptor systemfortheinvivorecombineeringofscFvintoIgGmolecules,JImmunolMethods,doi:10.1016/j.jim.2016.02.003.FerreiradeFreitas,R.,Eram,M.S.,Smil,D.,Szewczyk,M.M.,Kennedy,S.,Brown,P.J.,Santhakumar,V.,Barsyte-Lovejoy,D.,Arrowsmith,C.H.,Vedadi,M.,Schapira,M.(2016). DiscoveryofaPotentandSelectiveCoactivatorAssociatedArginine Methyltransferase1(CARM1)InhibitorbyVirtualScreening,JMedChem,doi: 10.1021/acs.jmedchem.6b00668.Li,M.,Yang,S.,Xu,D.(2016). HeparanSulfateRegulatestheStructureandFunctionof OsteoprotegerininOsteoclastogenesis,JBiolChem,doi: 10.1074/jbc.M116.751974.

Quality:

FectoPRO^®iscompliantwithbiomanufacturingguidelines.Indeed,FectoPRO^®isguaranteedfreeofanimal-origincomponents,andischemicallydefined.Systematiclotmanagementandreleasetestingisperformedforeachlotproduced.FurThermoreFectoPRO^®transfectionkitundergoesadvancedqualitycontrols:proteinproductivity,cellviability,andcompletesterility.

Testimonials:

“Inourhands,FectoPRO^®givesthehighestproteinproductionyieldsweeverreachedusingtransienttransfection.Ourcompanyofferscustomservicesincludingproteinproductioninmammaliancells.Beingabletoproducealotmoreproteinwithoutrevisitingourexpressionsystemopensawholenewrangeofpossibilitiesforourantibodyandproteinproductionservices”.
PhilippeF.,ProteoGenixSAS,France

“IhavetoadmitIhadlowexpectationsforFectoPRO^®.Perhapsamodestimprovementintransfection…anextra5-10%.J.didherfirstexperimentyesterdayandanalyzedittodayandtheresultwasremarkable.I’mshockedbyboththeefficiencyoftransfectionandthebrightnessoftheGFP.”
ThomasC.,Biotech,Cambridge,MA,UnitedStates

“2xtheyieldforhalftheamountoftransfectionreagent=4xlesscost. Itisalsoveryeasytouse.”
Jean-PhilippeJ.,HospitalforSickChildren,Canada

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Polyplus Transfection/FectoPRO®/Polyplus Transfection-161253/1 Ea