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1、VaccineNewaviansuspensioncelllinesprovideproductionofinfluenzavirusandMVAinserum-freemedia:Studiesongrowth,metabolismandviruspropagation.III新的禽流感懸浮細(xì)胞系提供生產(chǎn)的流感病毒和MVA無血清媒體:研究生長、新陳代謝和病毒傳播。1.IntroductionSeveralvaccineviruses‘includingmostofthehumaninfluenzavaccine
2、s,arestillproducedintheallantoiccavityofembryonatedhen'seggsorinprimarychickenfibroblasts(CEF).Maindraw-backsofthismethodarelimitedscalabilityoftheprocess,possibleshortageofmaterialincaseofanavianpandemicandoccasionaldifficultiesinamplificationofcertainstrain
3、swithrequiredseasonalantigenicity[l].ContinuouscelllinestoreplaceprimarymaterialforrobustindustrialproductionespeciallyofinfluenzaAandBarethereforehighlydesirable.MDCKandVerocellscanbeusedforproductionofvaccines.However,duetoinherentgeneticinstabilityofVeroce
4、llsaproductionprocessisusuallylimitedtoonly20pas-sages[2].Scale-upislimitedbytheirrequirementforanattachmentsurfaceandcultivationtypicallyisperformedinmediaContainingserum.Serumevenofpharmaceuticalgradesuffersfromlot-to-lotvariationsandisapotentialsourceforco
5、ntaminationwithadventitiousagents.Contrarytoinfluenzavaccines,modifiedvacciniaAnkaraviruses(MVA,recombinantorwildtype)arestronglyhostrestrictedsothatmostmammaliancells(includingMDCKandVerocells)arenotfullypermissive.ForthisreasonproductionofMVArequiresCEFcult
6、ures,againlimitingthescalabilityandrobustnessoftheprocess.MVAvectorsagainstcomplexinfectiousdiseaseswillberequiredathightitressothatefficientproductionprocessesarehighlydesirable.InordertoovercometheproblemsininfluenzaandMVAproductionnewcelllinesweredeveloped
7、recently[3-5].Developmentanddocumentationofthesecelllineswasperformedtomeetcurrentregulatoryrequirements.Thecelllinesweredesignedandoptimizedforaparticularproductionprocessandtypicallyproliferatewithanindefinitelifespaninsuspensioninserum-freemediumwithzeroor
8、lowproteincontent.ThebestknownexampleofadesignercellisthehumanPER.C6linewhichwasderivedfromaprimarycultureofhumanfetalretinoblastsimmortalizedbytransfectionwiththeElexpressioncassetteofad
