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酵母表达肽抗生素hPABβ的纯化及其抗单纯疱疹病毒活性分析
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酵母表达肽抗生素hPABβ的纯化及其抗单纯疱疹病毒活性
官方公共微信Optimization of microcarrier cell culture process for the inactivated39
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Optimization of microcarrier cell culture process for the inactivated39
Vaccine22(2004)；；Optimizationofmicrocarri；enterovirustype71vaccine；Suh-ChinWua,b,?,Chia-Chy；DepartmentofLifeScience,；cCenterforDiseaseControl；Received24May2002;accept；Avai
Vaccine22(2004)Optimizationofmicrocarriercellcultureprocessfortheinactivatedenterovirustype71vaccinedevelopmentSuh-ChinWua,b,?,Chia-ChyiLiua,Wei-ChengLiancaDepartmentofLifeScience,InstituteofBiotechnology,NationalTsingHuaUniversity,Hsinchu,TaiwanbVaccineResearchandDevelopmentCenter,NationalHealthResearchInstitutes,Taipei,TaiwancCenterforDiseaseControl,Taipei,TaiwanReceived24May2002;accepted21May2004Availableonline22July2004AbstractEnterovirus71(EV71)isanenterovirusthatcouldleadtosevereneurologicaldisordersandfatalities.TheinactivatedvaccineisanappropriateEV71vaccineformatformeetingcurrentneeds.Large-scalepreparationoftheinactivatedEV71vaccinedependsonascalablecellculturesystemforindustrialmassproduction.Inthispaper,VerocellswerefoundtoproducehighertitersofEV71thandidMRC-5andWI-38cells.High-densitymicrocarrierVerocellcultureswereestablishedusing5g/LCytodex1microcarriersandfoundtopromotethereleaseofEV71sfrominfectedVerocells.Forthelarge-scaleproductionoftheinactivatedvaccineantigen,theextracellularvirustitersproducedinthe2Lbioreactorwerefoundtobe10timeslowerthanthespinner?askculturebutimprovedby30-foldsusingglucose/glutaminefeedingsduringinfection.Aserum-freeVerocellmicrocarrierculturewasalsoestablishedinthebioreactor,yieldingahigh-titerof5.8×107TCID50/mLforEV71production.Theimmunogenicityoftheinactivatedvirionsproducedinserum-freecultureelicitedaslightlyhigherlevelofneutralizingantibodyresponseinimmunizedmice.Theseresultsconstitutevaluableinformationonthedevelopmentofalarge-scalemicrocarriercellcultureprocessforproducinginactivatedEV71vaccine.?2004ElsevierLtd.Allrightsreserved.Keywords:Enterovirus71;IMicrocarriercellculture1.IntroductionEnterovirus71(EV71)isanon-enveloped,single-strandedRNAvirusofthefamilyPicornaviridae,genusEnterovirus.TheEV71infectioninhumansmanifestsmostcommonlyasachildhoodexanthemaknownashand-foot-and-mouthdisease,buthasatendencytocauseneurologicaldiseaseduringacuteinfection[1].EV71was?rstisolatedin1969intheUnitedStates[2]andsubsequentlyreportedinAustralia[3],Sweden[4],Japan[5],Bulgaria[6],andHungary[7].Recently,EV71epidemicshaveoccurredinSarawak[8],Japan[9],peninsularMalaysia[10],Singapore[11],Taiwan[12],andAustralia[13].EV71epidemicsinTaiwancaused78deathsin1998,25deathsin2000,and26?Correspondingauthor.Fax:+.E-mailaddress:scwu@life.nthu.edu.tw(S.-C.Wu).deathsinC15].DevelopinganeffectivevaccineagainstEV71infectionisthebestwaytosolvethispublichealthproblem.Theonlyvaccineavailableagainstoneofawiderangeofenterovirusesisthatagainstpoliovirusinfection.Theinactivatedpoliovirusvaccine(IPV)developedbyJonasSalkwashighlyeffectiveinreducingtheincidenceratesofpoliomyelitisinthe1940sand1950s[16].Basedonthesameconcept,aninactivatedEV71vaccinewasdevelopedinresponsetotheBulgarianepidemicin1975[6],butnoef?-cacydatahavebeenobtainedsincetheydidnotoccuragain.LaboratorymicewererecentlydemonstratedtobeasuitableanimalmodelofEV71infection[17],althoughCynomolgusmonkeysarealsosusceptibletoinfection[18].Inlaboratorymice,inactivatedEV71vaccinehasbeendemonstratedtoelicitastrongerimmuneresponsethantherecombinantVP1proteinorDNAvaccines[19],indicatingtheinactivatedX/$Cseefrontmatter?2004ElsevierLtd.Allrightsreserved.doi:10.1016/j.vaccine.S.-C.Wuetal./Vaccine22(643859vaccineisanappropriateEV71vaccineformatformeetingcurrentneeds.Additionally,aVerocell-adaptedEV71strainwasdevelopedasaprototypevaccinewithhighviralyieldandbroadimmunogenicity[20].However,thelarge-scalepreparationoftheinactivatedEV71vaccinedependsonascalablecellculturesystemforindustrialmassproduction.Onlyafewcurrentcellsubstratesystemsareuseableinmanufacturinghumanvaccines.Acceptablecellsubstratesincludediploidcellstrains(MRC-5,WI-38,andFRhL-2)andcontinuouscelllines,suchasVero(Africangreenmonkeykidney)andMDCK(Modin-Darbycaninekidney)cells[21].However,theacceptabilityofthesecellsmustbeanchorage-dependent,sothesecellsmustadheretothesurfaceofamatrixtofacilitatecellgrowth.Microcarriertechnology[22]wasinitiallydevelopedforculturinganchorage-dependitcanbeeasilyappliedinstirred-itisscalable,anditseffectivenesseasilyvalidated.Microcarriercellculturesystemsoffermanyad-vantagesoverconventional?askorbottleculturesingrowinganchorage-dependentcellsandhavebeenusedfortheindus-trialmanufactureofhumanpolioandrabiesvaccines[23,24].ThisworkaddressestheproductionyieldsoftheEV71-1207straininculturedVero,MRC-5,andWI-38cells.High-densityVerocellcultureswerederivedbyincreasingthemicrocarrierconcentrationfrom2to5g/Land10g/LCytodex1.Thespeci?ctitersoftheintracellularandextracellularviruseswereobtainedtocomparethevialproductivityatvariousmicrocarrierconcentrations.A2L,bioreactorcultivationofaVerocellmicrocarrierculturewaspreformedinthelarge-scaleproductionoftheinactivatedvaccineantigen.Aserum-freeVerocellmicrocarrierculturewas?nallyestablishedinthebioreactor.Theimmunogenic-ityoftheinactivatedvirionsproducedinserum-freeculturewasexaminedfortheelicitationofneutralizingantibodyre-sponseinimmunizedmice.Theseresultsconstitutevaluableinformationonthedevelopmentofalarge-scalemicrocarriercellcultureprocessforproducinginactivatedEV71vaccine.2.Materalsandmethods2.1.Cells,media,andvirusVero,WI-38,andMRC-5cellswereobtainedfromtheCultureCollectionandResearchCenter,FoodIndustrialRe-searchandDevelopmentInstitute,Hsinchu,Taiwan.VerocellsweregrowninM199mediumsupplementedwith10%fetalbovineserum(FBS)(GIBCO,LifeTechnologies,US).WI-38andMRC-5cellsweregrowninDMEMmediumwith10%FBS.Rhabdomyosarcoma(RD)cellswereobtainedfromtheCenterofDiseaseControl,Taipei,Taiwan,andmaintainedinEagle’sminimumessentialmedium(MEM)plus10%FBS.AllofthesecellswerepassagedtwiceweeklyinT?asks.TheEV71-1207strainwasobtainedfromtheCen-terofDiseaseControl,Taipei,Taiwan.StocksofEV71-1207werecollectedfromthesupernatantsoftheinfectedRDcellsafterthreedayspostinfection(p.i.).Virusstockswerestoredat?80?Cfreezer.2.2.PreparationofmicrocarriersCytodex1microcarrierwaspurchasedfromAmershamBiosciencesandpreparedfollowingthemanufacture’sin-structions[25].Cytodex1microcarrierwasimmersedinphosphatebuffersaline(PBS)formorethan3hbeforeeachexperimentwasperformed.Afteritwasautoclavedfor15min,themicrocarriersatspeci?edconcentrationswerewashedtwicewiththeculturemedia.2.3.Spinner?askculturesHundredmLspinner?asks(Bellco)wereused,withaworkingvolumeof100mL.The?askswereplacedonamag-neticstirrerat60rpmina5%CO2incubatorat37?Cduringthecellgrowthphaseandat32?Cduringtheinfectionphase.CellsweredetachedfromT-?asksusingtrypsin-EDTA,andthentransferredtothespinner?asksatthegivencellandmi-crocarrierconcentrations.Cellsattachedtothemicrocarrierswerecountedbyusingthenucleistainingmethod[25].Thecellgrowthphasetestedforapproximately3days.Thespin-ner?askswereremovedtothelaminarhoodandsettledfor3mintoseparatethemicrocarriersandmediumthroughthegravitationalforce.Infectionwasthenperformedbyreplac-ing70%mediumwiththefreshmediumthatcontainedthestocksofEV71-1207inanamountthatcorrespondedtothemultiplicityofinfection(MOI)usedintheexperiments.2.4.CultivationinbioreactorandserumfreemediumABIOSTATbioreactor(B.BRAUN,Germany),inwhichasiliconmembranewasinsertedtoensurethatthegassup-plywasfreeofbubbles,wasusedforbioreactorcultivation.Thebioreactorculture(1Lworkingvolume)wasstirredatarateof60rpmatpH=7.2,controlledbythesurfaceaera-tionofCO2throughthegasmixerapparatus.TheVerocellsusedinthebioreactorexperimentswerecollectedfrom24T-150culture?asks.Themediumandmicrocarrierwereintro-ducedinitiallytothevesselandthentheinoculatedcellsweretransferredtothebioreactordirectlywithaconstantstirring(60rpm).Theinoculationcelldensityinthebioreactorcul-turewasaround5×105cells/mLandthecelldensityreached1.2C1.5×106cells/mLfollowing72hofcultivation.Infec-tionwasthenpreformedbyreplacing70%ofthemediumwiththefreshmediumthatcontainedthevirusstockatMOI=1.Inthebioreactor,theagitationwasstoppedfor5mintoseparatethemicrocarriersandmediumthroughthegravita-tionalforce.Theincubationtemperaturewasthenchangedto32?C.Glucose/glutamineconcentrates(?nalconcentrationof5.6mMglucoseand1.0mMglutamine)werefedtwicetoat12hp.i.and24hp.i.Themodi?edVeroserum-freemedium(M-VSFM)wasprovidedbyProfessorMikeButler,UniversityofManitoba,3860S.-C.Wuetal./Vaccine22(64Canada[26].VerocellswerepassageddirectlyintoM-VSFMfrom10%FBS-containingmedium199withoutanyadapta-tion.Inserum-freeculture,0.25%(w/v)trypsininhibitorwasusedfordetachingcellsduringcellpassagesandtransferringtobioreactor.VerocellsweremaintainedinM-VSFMforatleasttwopassagesinT-150?asksbeforethebioreactorcultivation.2.5.DeterminationofvirustiterVirustitersweredeterminedfromthemedianendpointofthetissueculture’sinfectiousdose(TCID50).TheintracelluarvirustiterwasdeterminedbymeasuringtheTCID50valuesoftheinfectedcelllysatestreatedoverthreefree-thawcycles.Theextracellularvirustiterwasmeasureddirectlyinculture?2supernatants.Seriallydilutedvirussamplesfrom10to10?9wereaddedtoVerocellsin96-wellplates.Quadraduplicatesampleswereusedateachdilution.Theculturesin96-wellplateswereincubatedforsevendaysat37?C,andtheTCID50valuesweremeasuredbycountingthecytopathiceffectsininfectedVerocells.TheTCID50valuewascalculatedusingthesoftware,ID-505.0,developedbyJ.Spouge,NationalCenterforBiotech-nologyInformation,Bethesda,MD,USA(http://www.ncbi.nlm.nih.gov/CBBresearch/Spouge/Virology/).2.6.PreparationofinactivatedEV71sandmouseimmunizationVirusstocksweremixedwitha37%formaldehydeso-lution(Merck)atavolumeratioof4000:1ratioandstoredat4?Cfor35to40daysforvirusinactivation.Theinacti-vatedvirusstockswereabsorbedonaluminumphosphateatroomtemperaturefor1handat4?Cfor1daybeforeimmu-nization.FemaleICRmice(12C15g,3C4weeksafterbirth)wereimmunizedintraperitoneally(i.p.)with0.5mLofthealum-absorbedinactivatedEV71s,andthesamedoseswereboostedi.p.after2weeksor3weeksafter?rstimmuniza-tion.Theimmunizedmicewerebled1weekaftertheboost,andserumsampleswereinvestigatedforvirusneutralization.The50%neutralizationinhibitiondose(Neut-ID50),whichisthegeometricreciprocaloftheserumdilutionyielding50%reductioninthevirustiterwasobtainedusingthesoftware,ID-505.0.3.Resultsanddiscussion3.1.SelectingcelllineforEV71productionThreemammaliancelllines(Vero,WI-38,andMRC-5)commonlyusedforpreparinghumanvaccineswereinves-tigatedfortheiruseinEV71production.These?cellsweregrowntocon?uencein6-wellplatesat37C(cell?growthphase)andinfectedwiththeEV71-1207strainat32C(virusinfectionphase)atMOI=1.TheintracelluarvirustiterFig.1.ThetitersofextracellularandintracellularEV71sproducedinthreemammaliancelllines(Vero,WI-38,andMRC-5).Sampleswerecollectedfromtheculturesupernatantsandcelllysatesat120hp.i.wasdeterminedfromtheTCID50valuesoftheinfectedcelllysatestreatedinthreefree-thawcycles.Theextracellularvirustiterwasdirectlymeasuredintheculturesupernatants.TheresultsindicatedthattheVerocellsgeneratedhighertitersofintracellularandextracellulartitersofEV71,rangingfrom2C5×107TCID50/mLoftheirmaximumtiterscollectedat72hp.i.,thandidWI-38andMRC-5cells(Fig.1).VerocellswerethuschosenasthecelllineforproducingEV-71.3.2.MicrocarriercellcultureforEV-71productionMicrocarriercellculturewasthenusedforproducingEV-71.VerocellsweregrownonCytodex1microcarriersatconcentrations2,5,and10g/Lin100mLspinner?asks.Thecelldensityforinoculationathighermicrocarrierconcen-trationswasincreasedfrom2×105cells/mL(2g/L)to5×105cells/mL(5g/L)and1×106cells/mL(10g/L).Un-dersuchconditions,thecelldensityoftheVerocellsgrownonmicrocarriers6reached1.1×106,1.2×106,and1.9×10cells/mL,respectively,following72hofinoculation(Fig.2).Microcarrier-growncellsweretheninfectedwithEV71(MOI=1)andcoupledwitha70%culturemediumreplacement.Theresultsrevealthattheextracellularvirustiters8of2and5g/Lmicrocarrierculturesexceededover1.0×10TCID50/mLafter150h(or3daysp.i.)(Fig.3A).Thereplicationkineticsoftheextracellularvirusesin10g/Lmi-crocarrierculturewasslower,andthemaximumextracellularvirustiterwaslowerat3.0×107TCID50/mL.Inreplicat-ingtheintracellularviruses,themaximumtitersof2,5,and10g/Lmicrocarriercultureswereallsimilar(Fig.3B).Thespeci?ctitersmeasuredpercell(TCID50/cell)wereobtainedtocomparethevirusyieldsproducedun-dervariousexperimentalconditions.Inextracellularvirusproduction,thespeci?cyieldsobtainedfromthemicro-carrierculturesincreasedfrom70TCID50/cell(2g/L)toS.-C.Wuetal./Vaccine22(643861Fig.2.ThegrowthcurvesforVerocellsonmicrocarriersusing2g/L(?),5g/L(??),and10g/L(??)Cytodex1in100mLspinner?asks.Thecellgrowthphasewasculturedat37?CtheninfectedwiththeEV71strain1207at32?C.Fig.3.Theviralreplicationkineticsfor(A)theextracellularvirusand(B)theintracellularvirusofEV71producedinVerocellmicrocarriercultures.VariousconcentrationsofCytodex1microcarrierswereused,including2g/L(?),5g/L(??),and10g/L(??)in100mLspinner?asks.120TCID50/mL(5g/L)butdeclinedto20TCID50/cell(10g/L)(Fig.4).Inintracellularvirusproduction,thespe-ci?cyieldsalsodeclinedasthemicrocarrierconcentrationin-creasedfrom5TCID50/cell(2g/L)to3TCID50/mL(5g/L)and2TCID50/cell(10g/L)(Fig.4).Additionally,ahigh-densityVerocellcultureof7×106cells/mLwasgeneratedtoproduceEV-71using15g/Lmicrocarrier(datanotshown),buttheresultantspeci?cyieldoftheextracellularvirustiterdeclinedfurtherto2TCID50/mL,about60timeslowerthanthatobtainedusing5g/Lmicrocarrier.Thedeclineinthespe-ci?cyieldsofEV-71forhigh-densitymicrocarrierculturesmayberelatedtothelimitedsupplyofnutrientsinculturesorotherfactors,asstatedelsewhere[27].Undertheexperimen-talconditionsherein,Verocellsgrownon5g/LmicrocarriersyieldedthehighesttitersofEV-71productioninspinner?askcultures.AnotherinterestingresultwasthatmicrocarrierVerocellculturespromotedthereleaseofEV-71particlesintoculturesupernatants.EV-71particleshavebeenpreviouslyreportedtoaccumulatemainlyintracellularlyascell-associatedviri-onsinaconventional?askorwell-plateculture[20].Theresultsobtainedusingawell-plateculturealsorevealedthatalargeproportionofEV-71particleswereproducedintracel-lularlyinVerocells(Fig.1).EV71isanon-envelopedRNAvirus,sotheproductionofamatureEV7virioniscompletedintracellularlywithouttheacquisitionofmembranesfromsubcellularorganellesorthecytoplasmofthehostcells.Theincreaseintheextracellularproductionofthevirusinmicro-carrierculturemayfollowsimplyfromthegreaterdamagetotheintegrityofthemembranebythehydrodynamicshearforces,whichthusfacilitatethereleaseofEV-91virions.However,thetriggeringofothercellularfactorsbyhydro-dynamicforcescannotbecompletelyruledout.MicrocarrierVerocellculturehastheadvantageofharvestingEV71par-ticlesintheculturesupernatants,insteadofintheinfectedcells,simplifyingdownstreamprocessinginthedevelopmentofinactivatedEV-71vaccines.3.3.Microcarrierbioreactorculture3.3.1.Serum-containingmediumwithoutfeedingduringinfectionA2-Lbench-topbioreactorwasusedtocultureVerocellsgrownon5g/Lmicrocarrierstoscale-upthecellculturepro-cessforEV-71production.ThegrowthcurveofVerocellsonmicrocarriersinthebioreactorreachedamaximumcellden-sityof1.3×106cells/mLfollowing72hinoculation,andthecellsweretheninfectedwithEV-71atMOI=1(coupledwitha70%mediumexchange)(Fig.5A).ThereplicationcurvesofEV-71producedinthebioreactorcultivationwereplottedastitersofextracellularandintracellularvirusesateachspec-i?edtime(Fig.5A).Theseresultsrevealthattheextracelluarvirustitersincreasedfrom1×106TCID50/mLat72h(or0hpost-infection)to3.0×107TCID50/mLat192h(or120hpost-infection)inthebioreactorculture.Themaximumex-tracellularvirustiterwasaboutthentimeslowerthanthatofthe100mLspinner?askculturesatthesameconcentrationofmicrocarriers(5g/L),althoughtheintracellularvirustiters(3.1×106TCID50/mLinthebioreactorcultureversus4.5×106TCID50inthespinner?askculture)andthecelldensi-tiesduringinfectionweresimilar(1.3×106cells/mLinthe3862S.-C.Wuetal./Vaccine22(2004)Fig.4.Thespeci?cvirustitersofEV71producedinVerocellmicrocarriercultures.Thespeci?cvirustitersareindicatedintheextracellularvirustiter(?)andintracellularvirustiter(??).bioreactorcultureversus1.2×106cells/mLinthespinner?askculture).Thelowyieldofextracellularvirusproducedinthebiore-actorcultivationmayberelatedtothestrongerhydrodynamicshearforcesinthebioreactor.Microcarrier-attachedcellsarebelievedtobemoresensitivetohydrodynamicshearforcesthansinglecellsinsuspension,becausetheyareanchoredtomicrocarriersurfaceandsocannotfreelyrotateortranslatetoreducethenetforcesthatexertedonthem[28].Thelackofasigni?cantdeclineintheintracellularvirusproductioninthebioreactorindicatesthatthereducedextracellularviruspro-ductioninthebioreactorcultivationisprobablynotrelatedtodamagetotheintegrityofthemembraneoftheinfectedcells,anditscontributiontovirusrelease,ortoincreasecul-turevolumeinthebioreactorvesselandtheconsequentlylessef?cientadsorptionofthevirus.3.3.2.Serum-containingmediumwithfeedingsduringinfectionTheinfectedcellswerefedwithaglucose/glutaminecon-centrateduringtheinfectionphasetoincreasefurthertheextracellularproductionofvirusinthebioreactorculture(Fig.5B).Twofeedingsofglucoseandglutamineat12and24hpost-infectionmaintained&2C4mMglucoseintheculturemediumthroughouttheinfectionperiod(datanotshown).Theresultedrevealthatthebioreactorculture,withthefeedingstrategy,exhibitedanincreasedextracellularvirustiterof8.9×107TCID50/mL(Fig.5B).InintracellularEV71production,nosigni?cantincreaseofthemaximumtiterswasfoundinthebioreactorculturewithglucose/glutaminefeedings(3.2×106TCID50/mLwithfeedingsverse3.1×106TCID50/mLwithoutfeeding).Oneexplanationisthatonlyaverysmallfractionbyaround3%oftheEV71swasproducedintracellularlyfromthemicrocarrier-grownVerocells,sothenutrientfeedingsdidnotyieldasigni?cantin-creaseoftheintracellularvirusproduction.3.3.3.Serum-freemediumwithfeedingsduringinfectionTheserum-freemicrocarrierculturewasalsofedwithglucose/glutamineusingM-VSFMserum-freemediumtogrowVerocells[26,27,29].TheresultsindicatethattheVerocellgrowthonmicrocarriersusingM-VSFMserum-freemediumwassimilartothatusingM199mediumsupple-mentedwith10%FBS(Fig.5C).Themaximumextracellularvirustiterproducedintheserum-freebioreactorculturewas5.8×107TCID50/mL,30%lowerofthevirusyieldobtainedintheserum-supplementedbioreactorculturewithglucose/glutaminefeedings.Themaximumintracellularvirustiterproducedintheserum-freebioreactorculturewas9.0×106TCID50/mLthatarearoundtwo-foldshigherthanthemaximumtitersobtainedinserum-supplementedbioreactorculture.ThedifferencesmaybeduetosomecomponentsintheserumfreemediumM-VSFMaffectingtheviralassemblyandreleaseininfectedVerocells.Furtheroptimizationofthefeedingcomponentsandscheduleswillimprovetheproductionprocessinserum-freeVerocellculture.3.4.Immunogenicityofformalin-inactivatedEV71sharvestedfromserum-freeandserum-supplementedculturesTofurtherdeterminewhetherserum-freecultureaffectstheimmunogencityofEV71virions,thevirusstocks包含各类专业文献、行业资料、各类资格考试、高等教育、专业论文、应用写作文书、外语学习资料、生活休闲娱乐、Optimization 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&&Dell Networking S-Series Managed 10/40Gb Ethernet Switches&&&
Legend: S — Standard, N — Not AvailablePerformanceS4810S4820TS5000S6000MAC Address128,000128,000128,000160,000IPv4 routes16,00016,00016,00016,000IPv6 routes8K (shared CAM space with IPv4)8K (shared CAM space with IPv4)8K (shared CAM space with IPv4)8K (shared CAM space with IPv4)Switching fabric capacity1.28 Tbps (full-duplex)640 Gbps (half-duplex)1.28 Tbps (full-duplex)640 Gbps (half-duplex)1.28 Tbps (full-duplex)640 Gbps (half-duplex)2.56 Tbps (full-duplex)1.28 Tbps (half-duplex)User traffic capacity960 Mpps960 Mpps960 Mpps1462 MppsLink aggregation8 links per group, 128 groups per stack8 links per group, 128 groups per stack8 links per group, 128 groups per stack8 links per group, 128 groups per stackStacking capacity160Gbps160Gbps160Gbps160GbpsQueues per port4 queues4 queues4 queues8 queuesVLANs4,0004,0004,0004,000Line-rate Layer 2 switching — All protocols, including IPv4 and IPv6SSSSLine-rate Layer 3 routing — IPv4 and IPv6SSSSLAG load balancing — Based on Layer 2, IPv4 or IPv6 headersSSSSSwitching latency800ns3.3us800ns600nsPacket buffer memory9MB9MB9MB12MBCPU memory2GB2GB2GB4GBIPv4 host table size4,0008,0008,00048,000IPv6 host table size4,0008,0008,00032,000IPv4 Multicast table size4,0008,0008,00064,000
AvailabilityS4810S4820TS5000S6000Ring stacking topology with dynamic master electionSSSNDual modular slots with up to four 10 GbE portsSSSNLink aggregation across stack membersSSSSBuilt-in power redundancySSSSHot swappable redundant AC or DC powerSSSSHot swappable redundant fansSSSSChassisS4810S4820TS5000S6000Line-rate ports48 - 10 Gigabit Ethernet SFP+ ports48 - 10 GBASE-T ports48 - 10 Gigabit Ethernet SFP+ ports & FC/FCoE ports32 - 40 Gigabit Ethernet QSFP+ ports or up to 96 - 10 GbE SFP+ & 8 - 40GbE ports (breakout mode)SFP Ports4 - 40 Gigabit Ethernet QSFP+ ports4 - 40 Gigabit Ethernet QSFP+ ports4 - 40 Gigabit Ethernet QSFP+ ports32 - 40 Gigabit Ethernet QSFP+ ports1 RJ45 console/management port with RS232 signalingSSSS1 RJ45 Ethernet management portNNNSUSB PortsNNSS2 line-rate ports 10 Gigabit Ethernet XFP ModulesNNNN2 line-rate ports 10 Gigabit Ethernet CX4 ModulesNNNN2 line-rate ports 12 Gigabit Stacking ModulesNNNN2 line-rate ports 10 Gigabit Ethernet SFP+ ModulesNNNN1 line-rate port 24 Gigabit Stacking ModulesNNNNDimensions S4810S4820TS5000S6000Size1 RU, 1.73 h x 17.32 w x 18.11" d (4.4 h x 44 w x 46 cm d)1 RU, 1.71 h x 17.09 w x 18.11" d (4.35 h x 43.4 w x 46 cm d)1RU, 1.73 h x 17.32 w x 28" d (4.4 h x 44 w x 46 cm d)RU, 1.73 h x 17.32 w x 18.11" d (4.39 h x 43.99 w x 46 cm d)Weight14.39 lbs (6.54 kg)21.7 lbs (9.86 kg)34 lb (15.42 kg)14.41 lbs (6.54 kg)ISO 7779 A-weighted sound pressure level59.6 dBA at 73.4°F (23°C)65 dBA at 78.8°F (26°C)59.6 dBA at 73.4°F (23°C)20 dBA at 73.4°F (23°C)Environmental S4810S4820TS5000S6000Operating temperature:32° to 122°F (0° to 50°C)32° to 104°F (0° to 40°C)32° to 113°F (0° to 45°C)32° to 104°F (0° to 40°C)32° to 113°F (0° to 45°C)Operating humidity:10 to 85% (RH), noncondensing10 to 85% (RH), noncondensing10 to 90% (RH), noncondensing10 to 85% (RH), noncondensing10 to 90% (RH), noncondensingStorage temperature:–40° to 158°F (–40° to 70°C)SSSSStorage humidity:5 to 95% (RH), noncondensingSSSSPowerS4810S4820TS5000S6000Power supply100–240 VAC 50/60 Hz100–240 VAC 50/60 Hz100–240 VAC 50/60 Hz-48 to -60 VDC100–240 VAC 50/60 HzMax. thermal output:955.36 BTU/h1433 BTU/h1878 BTU/h1265.9 BTU/hMax. current draw per system2 A at 100/120 VAC, 1 A at 200/240 VAC4.2A at 100/120VAC, 2.1A at 200/240VAC, 1.04A at 40.5VDC, 7A at 60VDC 7A at 100/120VAC, 3.5A at 200/240VAC, 15.2A at -46VDC, 11.7A at -60VDC 3.8A at 100/120 VAC, 1.6A at 200/240 VACMax. power consumption350 Watts460 Watts550 Watts371 WattsMax. PoE powerNNNNReliabilityNNNNSafetyS4810S4820TS5000S6000EN 60825-1 Safety of Laser Products Part 1: EquipmentSSSSEN 60825-2 Safety of Laser Products Part 2: Safety ofSSSSEN 60950-1, Second EditionSSSSEquipment Classification Requirements and User's GuideSSSSFDA Regulation 21 CFR 1040.10 and 1040.11SSSSIEC 60950-1, Second Edition Including all NationalSSSSOptical Fibre Communication SystemsSSSSUL/CSA 60950-1, Second EditionSSSSEmissions S4810S4820TS5000S6000Australia/New Zealand: AS/NZS CISPR 22: 2009, Class ASSSSCanada: ICES-003, Issue-4, Class ASSSSEurope: EN 5+A1:2007 (CISPR 22: 2006), Class ASSSSJapan: VCCI V3/2009 Class ASSSSUSA: FCC CFR 47 Part 15, Subpart B:2009, Class ASSSSImmunity S4810S4820TS5000S6000EN 300 386 V1.4.1: 2008 EMC for Network EquipmentSSSSEN 5 + A1: 2001 + A2: 2003SSSSEN : Harmonic Current EmissionsSSSSEN :Voltage Fluctuations and FlickerSSSSEN : ESDSSSSEN : Radiated ImmunitySSSSEN : EFTSSSSEN : SurgeSSSSEN : Low Frequency Conducted ImmunitySSSSRoHS S4810S4820TS5000S6000All S Series components are EU RoHS compliant.SSSSRegulatory information: (in English) (in English) (in English)
Legend: S — Standard, N — Not AvailablePerformanceS55S60MAC Address32K32KIPv4 routes16,00016,000IPv6 routes8,0008,000Switching fabric capacity192 Gbps176 GbpsUser traffic capacity144 Mpps132 MppsLink aggregation8 links per group, 128 groups per stack8 links per group, 128 groups per stackStacking capacity48 Gbps per stack member96 Gbps per stack memberQueues per port4 queues4 queuesVLANs with 4096 tag value support10241024Line-rate Layer 2 switching — All protocols, including IPv4 and IPv6SSLine-rate Layer 3 routing — IPv4 and IPv6SSLAG load balancing — Based on Layer 2, IPv4 or IPv6 headersSSSwitching latency<5 us for 64 byte frames<9 us for 64 byte framesPacket buffer memory4MB1.25GBCPU memory2GB2GBIPv4 host table sizeN/AN/AIPv6 host table sizeN/AN/AIPv4 Multicast table sizeN/AN/A
AvailabilityS55S60Ring stacking topology with dynamic master electionSSDual modular slots with up to four 10 GbE portsSSLink aggregation across stack membersSSBuilt-in power redundancySSHot swappable redundant AC or DC powerSSHot swappable redundant fansNSChassisS55S60Line-rate ports44 - 10/100/1000 Base-T ports44 RJ45 10/100/1000 Base-T portsSFP Ports441 RJ45 console/management port with RS232 signalingSS1 RJ45 Ethernet management portNNUSB Ports2 USB 2.0 ports (1 USB A, 1 USB B)2 USB 2.0 ports (1 USB A, 1 USB B)2 line-rate ports 10 Gigabit Ethernet XFP ModulesNN2 line-rate ports 10 Gigabit Ethernet CX4 ModulesNN2 line-rate ports 12 Gigabit Stacking ModulesSN2 line-rate ports 10 Gigabit Ethernet SFP+ ModulesSS1 line-rate port 24 Gigabit Stacking ModulesNSDimensionsS55 S60 Size1 RU, 1.7 h x 17.32 w x 18.06" d (4.3 h x 44 w x 45.87 cm d)1 RU, 1.7 h x 17.32 w x 20.5" d (4.3 h x 44 w x 52.07 cm d)Weight14.39 lbs (6.54 kg)14.39 lbs (6.54 kg)ISO 7779 A-weighted sound pressure level59.6 dBA at 73.4°F (23°C)59.6 dBA at 73.4°F (23°C)Environmental S55S60Operating temperature:SS32° to 122°F (0° to 50°C)SSOperating humidity:SS10 to 85% (RH), noncondensingSSStorage temperature:SS–40° to 158°F (–40° to 70°C)SSStorage humidity:SS5 to 95% (RH), noncondensingSSPowerS55S60Power supply100–240 VAC 50/60 Hz, –44 to -60 VDC100–240 VAC 50/60 Hz, –44 to -60 VDCMax. thermal output:443 BTU/h531 BTU/hMax. current draw per system2 A at 100/120 VAC, 1 A at 200/240 VAC, 3.6 A at -48 VDC2 A at 100/120 VAC, 1 A at 200/240 VAC, 3.6 A at -48 VDCMax. power consumption130 WNMax. PoE powerNNReliabilityMTBF 169,315 hoursMTBF 169,315 hoursSafetyS55S60EN 60825-1 Safety of Laser Products Part 1: EquipmentSSEN 60825-2 Safety of Laser Products Part 2: Safety ofSSEN 60950-1, Second EditionSSEquipment Classification Requirements and User's GuideNNFDA Regulation 21 CFR 1040.10 and 1040.11SSIEC 60950-1, Second Edition Including all NationalSSOptical Fibre Communication SystemsNNUL/CSA 60950-1, Second EditionSSEmissions S55S60Australia/New Zealand: AS/NZS CISPR 22: 2006, Class ASSCanada: ICES-003, Issue-4, Class ASSEurope: EN 5 (CISPR 22: 2006), Class ASSJapan: VCCI V3/2007.04 Class ASSUSA: FCC CFR 47 Part 15, Subpart B, Class ASSImmunity S55S60EN 300 386 V1.3.3: 2005 EMC for Network EquipmentSSEN 5 + A1: 2001 + A2: 2003SSEN : Harmonic Current EmissionsSSEN :Voltage Fluctuations and FlickerSSEN : ESDSSEN : Radiated ImmunitySSEN : EFTSSEN : SurgeSSEN : Low Frequency Conducted ImmunitySSRoHS S55S60All S Series components are EU RoHS compliant.SSRegulatory information:
Designed for performanceDell Networking switching technology enables outstanding productivity for virtualized data center and cloud computing environments. The wide range of S-Series switches includes the SG high-density top-of-rack (ToR) switch, a compact 1RU fixed switch with full L2/L3 capability, including Data Center Bridging (DCB) and Virtual Link Trunking (VLT).
Added flexibility to configure 32 x 40GbE ports or 96 x 10GbE plus 8 x 40GbE portsLow latency with superb performance and high density with hardware and software redundancyDCB for reliable, high-performance Ethernet in large data center networksStreamlined connectivity and workflow operation across physical and virtual resources with multi-tenancy capabilities Support for Network Virtualization with VXLAN Gateway hardware support integrated into the switchIntegrated automation, scripting and programmatic management for enhanced network flexibility in virtualized environments
Developed for flexibilityThe Dell Networking S-Series switches offer a family of products that enable you to find the ideal solution for your specific needs.Deliver versatility with line-rate performance and port densities from 48 x 1/10Gb Base T line-rate ports in the S4820T switches to 32 x 40GbE ports in the S6000Scale quickly with efficient stacking offered in many of the S-Series models, including ST/SGb) platformsPowered by our industry-hardened and feature-rich operating system for maximum dependability and uptimeFull support for iSCSI storage area networks including support for DCB
Built for cost-effective deploymentSelect S-Series Managed 10/40 Gb switches are supported with the Dell Open Automation Framework, a suite of interrelated network management tools that can be used together or independently to provide a flexible, available and manageable network while helping reduce operational expenses. The Open Automation Framework takes full advantage of Dell FTOS software capabilities to bring network automation into virtual data center environments. This helps the S-Series switches efficiently respond and adapt to changes in application requirements.
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Dell Networking SGbE ToR SwitchThe S6000 supports top-of-rack (ToR), middle-of-row (MoR) and end-of-row (EOR) connectivity to 10GbE servers and 40GbE fabrics for enterprise and midmarket implementations. The 1U S6000 enables streamlined connectivity and workflow operation with low latency operation and includes support for Network Virtualization with VXLAN Gateway hardware built-in. Virtual Link Trunking (VLT) helps eliminate spanning tree protocol (STP)-blocked ports and provides fast convergence if a link or device failsDell Networking SG Unified Storage SwitchThe S5000 enables converging LAN and SAN traffic in a single 1U top-of-rack switch. It features a unique modular design to enable adding Ethernet and Fibre Channel/FcoE modules when needed, and DCB support for lossless Ethernet. In addition, the S5000 helps standardize and simplify the Dell Active System Infrastructure pre-integrated systems.Dell Networking S4820T 10G Base T SwitchDeploy 1/10GbE for 10G copper terminations to server with the S4820T 10G Base T switch. The high-density switch features 48 ports of 1/10GbE and four 40GbE uplinks in a 1U rack chassis. Powered by the Dell Networking Operating System (FTOS), it offers non-blocking, line-rate performance for top-of-rack storage solutions and Dell 12G rack servers as well as 12G blade servers with Dell Networking MXL switches.Dell Networking S4810 High-Performance 10/40GbE SwitchDeliver low-latency throughput with the S4810. The 1/10/40GbE top-of-rack, aggregation or core switch features a 1.28 Tbps (full-duplex), non-blocking, cut-through switching fabric designed to deliver line-rate performance under full load with 800ns latency.Dell Networking S60 High-Performance 1/10GbE SwitchOptimized to improve manageability at the network edge, the S60 is a stackable, 48-port GbE top-of-rack switch that delivers non-blocking, line-rate switching and routing with a 1.25GB deep-packet buffer to help eliminate congestion.Dell Networking S55 High-Performance 1/10GbE SwitchDesigned for high-performance data center applications, the S55 features a 176 Gbps switch fabric that supports line-rate, non-blocking switching with less than five microseconds of latency.
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The S4820T earns 5/5 stars.“Crams features into its 1U chassis.” — ITPro