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1、High-DensityMicrocarrierCellCulturesforInfluenzaVirusProductionA.Bock,J.Schulze-Horsel,J.Schwarzer,E.Rapp,andY.GenzelMaxPlanckInstituteforDynamicsofComplexTechnicalSystems,BioprocessEngineering,39106Magdeburg,GermanyU.ReichlOtto-von-Guericke-Universita¨tMagdeburg,ChairofBio
2、processEngineering,Universita¨tsplatz2,39106Magdeburg,GermanyMaxPlanckInstituteforDynamicsofComplexTechnicalSystems,BioprocessEngineering,39106Magdeburg,GermanyDOI10.1002/btpr.539PublishedonlineJanuary7,2011inWileyOnlineLibrary(wileyonlinelibrary.com).InfluenzavirusA/PR/8/3
3、4viruspropagationinadherentMadin–Darbycaninekidneycellsinhigh-densitymicrocarrierculturesisdescribed.Toimprovevirusyields,perfusionandrepeatedfed-batchmodeswereappliedusingcell-specificfeedrates.Celldensitiesupto1.1710cells/mLwereachieved.Cell-specificvirusyieldsinhigh-dens
4、itycultureswereatsimilarlevelscomparedwithstandard,low-densitycultivations.Intheaverage2,400and3,300virionspercellwereobtainedfortwovariantsofthevirusstrainA/PR/8/34,PR8-NationalInstituteforBiologicalStandardsandControl(NIBSC)andPR8-RobertKochInstitute,respectively.Maximum
5、virustiter(HAactivity¼1,778HAU/100lL)forvirusvariantPR8-NIBSCwasobtainedforacultivationinfectedbeforemaximumcellconcentrationwasreached.VC2011AmericanInstituteofChemicalEngineersBiotechnol.Prog.,27:241–250,2011Keywords:high-densitycellculture,MDCK,microcarrier,influenza,vac
6、cineIntroduction‘‘celldensityeffect,’’firstdescribedbyWoodetal.(1982),hasbeenrepeatedlyusedoverthelastyearstodescribeFacingtheprobabilityofpandemicoutbreaksofinfluenzalowerthanexpectedcell-specificvirusyields(Lindsayandvirusandtheworld-widecostsrelatedtoannualhumanBetenbaugh,
7、1992;Marangaetal.,2003;Wongetal.,1996).influenzaAepidemics,significanteffortsarecurrentlymadePartofthis‘‘celldensityeffect’’couldberelatedtolimita-toincreasecapacitiesandyieldsininfluenzavaccinemanu-tionofmediumcompoundsoraccumulationofgrowthinhib-facturing.Astheconventionalp
8、roductioninhen’seggscan-itorslikeammoniaandlactate.Thesefactorscan,innotcopewiththedemand