产品资料

Polyplus Transfection/PEIpro^®/Polyplus Transfection-163149/1 Ea

OptimizedforLargeScaleTransientGeneExpressionforvirusproduction
IncreasedviraltiterscomparedtootherPEIs
Perfectlysuitableforthedevelopmentofbioproductionprocesses
Freeofanimal-origincomponents
WellcharacterizedandlotreleasetestedlinearPEI
Reliableandsecuresupply
Canalsobeusedforlargescaleproteinproduction

Specifications:

Reagent	PEIpro^®
Moleculedelivered	DNA
Applications	Virusproduction Proteinproduction
Celltypes	AdherentHEK-293andderivativesgrowninserumcontainingmedium SUSPensionHEK-293andderivativesgrowninsyntheticserum-freemedium OthercelllinesusedforvirusproductionsuchasVERO,CHOandBHKcells
Numberoftransfections	100mLofPEIpro^®transfectionreagentissufficienttotransfectonaverage50Lofcellculture
Storage	4°Cforupto2years

Summary:

Focusonyourproductionyield,useaPEIyoucanrelyon!

Orderinginformation:

|||115-010##10ml##|||115-100##100ml##|||115-400##4x100ml##|||

Description:

OptimizedfortransfectioncomparedtootherPEIs

ThelinearformofPEIpro^®andthemanufacturingprocessdevelopedbyPolyplus-transfection^®ensureahigh,stableandreproducIBLeamountofprotonableaminesavailablefortransfectionwhileprovidingatrulydeacylatedmoleculeandanextremelylowpolymerchainlengthvariation.Asaresult,PEIpro^®ismorepowerfulasitrequireslessDNAandreagentthananyotherlinearPEI(Fig.1)forincreasedvirusandproteinproductionyieldinHEK-293andCHOcellsgrowninserumfreemedia.

PEIpro - Comparison PEIs — **Fig.1:PEIpro^®requireslessreagentandsimilartolowerDNAamountcomparedtootherPEIs.**SuspensionHEK-293cellswereseededat1×10⁶cells/mlinserumfreemediumandtransfectedwithPEIpro^®,PEI“Max”andL-PEI25kDa(**polysciences**,Warrington,PA)resuspendedat1mg/ml.Luciferaseexpressionwasassayed48haftertransfectionusingaconventionalluciferaseassay.

PEIpro^®hasbeenformulatedat1mg/mltofitcurrentstandardsfortransfectionwithPEI.Thisconcentrationsimplifiesthetransfectionprotocoloptimization. PleaseContacttheTechnicalSupportforanyquestionsabouttheprotocol.

Wellcharacterizedwithlotreleasetesting

PEIpro^®isfullysyntheticandisfreeofcomponentsofanimal-origin.

ThequalityofPEIpro^®iscontinuouslyassessedduringthemanufacturingprocesswithappropriatecontroltesting.Systematiclotmanagementandreleasetestingisperformedforeachlotproduced.AstandardizedtransfectionassayisperformedwithsuspensionHEK-293celllineinserum-freemediatoensurethereliABIlityandreproducibilityofthetransfectionefficiencyandproteinyieldsunderconditionsthataresuitableforBiomanufacturing.

PEIpro^®undergoesstringentqualitycontrolstoensurecompletesterility,absenceofmycoplasmaandverylowlevelofbacterialendotoxinsfollowingpharmacopoeiastandardsandissuppliedwithadequatedocumentation.AfulllistofQCsisavailableuponrequest(ContacttheTechnicalSupport).Forproductionofclinicalbatchesofvirusandprotein,Polyplus-transfection^®offersahighlyqualifiedgradeofPEIpro^®,PEIpro^®-HQ.

Greatlottolotconsistency

PEIpro^®ismanufacturedandformulatedusingahighlycontrolledandreproducibleproductionprocess.Byassuringreproducibilityfromoneruntothenext,irrespectiveoftheoperator,PEIpro^®greatlyfacilitatesprocessstandardization;akeyadvantageforlarge-scalevirusandproteinproduction.Moreover,PEIpro^®isreleasedusingadvancedqualitycontrolsincludingaspecificationfortransfectionefficiencythatguaranteesexcellentlot-to-lotconsistency(Fig.2).

PEIpro - reproductibility IgG3 — **Fig.2:Excellentlot-to-lotproteinyieldreproducibilityusingPEIpro^®**.SuspensionHEK-293cellswereseededat1×10⁶cells/mlinserum-freemediumandtransfectedwithPEIpro^®followingthestandardprotocol.IgG3-Fcproductionwasassayed48haftertransfectionusingproteinGaffinityquantification(HPLC).

Versatileandeasytoscaleup

WithPEIpro^®,transfectionprocessesareeasytoscaleup.PEIpro^®canbeusedfromR&Dtolargescalevirusorproteinproduction.Witha given2yearshelflifeandareadytouseformulation,PEIpro^®isthereagentofchoiceforvirusproductionrunsinmostcellculturemediaandformat (Table1)usingbothadherentandsuspensioncells.

PEI - Virus yields compared with different culture media — **Table1:VirusyieldsusingPEIpro^®indifferentculturemediaforHEK-293cells.**

Applications:

Virusproduction

Withitsuniquepropertytotightlycondenseseveraldifferentplasmids,PEIpro^®isidealfortheco-transfectionofmultipleplasmidsthatisoftenrequiredforvirusproduction.

TransfectionwithPEIpro^®ensureshighviraltitersinmostcommonlyusedvirusproducingandpackagingcelllines(HEK-293,VERO,CAP,CAPT,etc.).PEIpro^®iswellsuitedforproductionofalltypesofviruses,achievingforexampleviraltiters>10⁷IG/mlforlentivirusesand>10¹¹to10¹²VG/mlforAAV(Fig.1).

PEIpro - Lentivirus and AAV production — **Fig.1:LentivirusandAAVproductioninHEK-293TandHEK-293cellsgrowninsuspensioninBalanCD^®HEK293(**Irvine**Scientific^®)**.HEK-293T(lentivirus)andHEK-293(AAV)cellswerethaweddirectlyintoeachmediumandpassagedevery3to4daysbeforegoingintoa2Lbenchtopbioreactor.Cellswereseededandculturedfor3daysbeforebeingtransfectedbyPEIpro^®(Polyplus-transfection^®).Fortransfection,fourplasmidswereusedforlentivirusandthreeplasmidswereusedforAAV.LentiviralandAAVtiterweremeasured48and72hoursposttransfection(DatakindlyprovidedbyGenethon).

ComparedtoCaPO4,PEIpro^®ismorerobustwithtransfectionefficienciesrangingfrom70to90%bothinadherentandinsuspensioncells.ItisalsomorepowerfulasitgiveshigherviraltitersusinglowerDNAamount(Fig.2.).

PEIpro - Virus titers — **Fig.2:PEIpro^®giveshighervirustitersthanCaPO4**.LentiviruseswereproducedinadherentHEK-293cellsgrowninserum-freeculturemedium,using15µgDNAand30µlPEIpro^®per75cm2flask.Viraltitersweredeterminedbyflowcytometryofsupernatants48haftertransfection.

PEIpro^®isparticularlysuitedforvirusproductionusingFreeStyle™F17medium(LifeTechnologies™).

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Proteinproduction

PEIpro^®isperfectlysuitedforlargescaleproteinproductioninbothHEK-293andCHOcells.Indeed,thisreagentiscompatiblewithvariousmediaformulationsforbothCHOandHEK-293cells.

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Citations:

HereisaselectionofrelevantreferencesusingPEIpro®,moreareavailableinourPolyplus-transfectionDatabase.Melidoni,A.N.,Dyson,M.R.,McCafferty,J.(2016). SelectionofAntibodiesInterferingwithCellSurfaceReceptorSignalingUsingEmbryonicStemCellDifferentiation. MethodsMolBiol1341,111-32.Stockinger,L.W.,Eide,K.B.,Dybvik,A.I.,Sletta,H.,Varum,K.M.,Eijsink,V.G.,Tondervik,A.,Sorlie,M.(2015). Theeffectofthecarbohydratebindingmoduleonsubstratedegradationbythehumanchitotriosidase. BiochimBiophysActa1854,1494-501.Bodnar,E.,Raymond,C.,Lopez,P.G.,Villacres,C.,Butler,M.,Schoenhofen,I.C.,DuRocher,Y.,Perreault,H.(2015). Massspectrometricanalysisofproductsofmetabolicglycanengineeringwithazido-modificationofsialicacids. AnalBioanalChem407,8945-58.Li,S.K.,Abbas,A.K.,Solomon,L.A.,Groux,G.M.,DeKoter,R.P.(2015). Nfkb1activationbytheE26transformation-specifictranscriptionfactorsPU.1andSpi-BpromotesToll-likereceptor-mediatedsplenicBcellproliferation. MolCellBiol35,1619-32.Fuller,J.P.,Stavenhagen,J.B.,Christensen,S.,Kartberg,F.,Glennie,M.J.,Teeling,J.L.(2015). Comparingtheefficacyandneuroinflammatorypotentialofthreeanti-abetaantibodies. ActaNeuropathol130,699-711.

Quality:

PEIpro^®isfullysyntheticandfreeofanyanimal-origincomponent.ThequalityofPEIpro^®iscontinuouslyassessedanddocumentedduringmanufacturingwiththeappropriatecontroltestings.Furthersystematiclotmanagementandreleasetestingisperformedforeachlotproduced.Aqualifiedtransfectionassayensuresreliabilityandreproducibilityofthetransfectionefficiencyunderconditionssuitableforbiomanufacturing.PEIpro^®undergoessterility,mycoplasmaandbacterialendotoxintestingaccordingtopharmacopoeiastandards.AfulllistofQCsisavailableuponrequest(ContacttheTechnicalSupport).

Testimonials:

GenethonproudlyusesPEIproforproductionofpre-clinicalbatchesofLentivirus&AAV.

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Polyplus Transfection/PEIpro®/Polyplus Transfection-163149/1 Ea