禽流感病毒的悬浮培养.doc

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1、VaccineNewaviansuspensioncelllinesprovideproductionofinfluenzavirusandMVAinserum-freemedia:Studiesongrowth,metabolismandviruspropagation.III新的禽流感悬浮细胞系提供生产的流感病毒和MVA无血清媒体:研究生长、新陈代谢和病毒传播。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