JournalofVisualizedExperimentswww.jove.comCopyright©2011JournalofVisualizedExperimentsSeptember2011|55|e3010|Page1of5VideoArticleAHigh-throughputAutomatedPlatformfortheDevelopmentofManufacturingCellLinesforProteinTherapeuticsShuangpingShi1,RussG.G.Condon1,LiangDeng1,JasonSaunders1,FinnHung1,Yung-ShyengTsao1,ZhongLiu11MerckResearchLaboratory,Merck&Co.,IncCorrespondenceto:ShuangpingShiatshuangping.shi@merck.comURL:http://www.jove.com/video/3010DOI:doi:10.3791/3010Keywords:Medicine,Issue55,Manufacturingcellline,proteintherapeutics,automation,high-throughput,FACS,FACSAria,CloneSelectImager,TECANFreedomEVOliquidhandlingsystemDatePublished:9/22/2011Citation:Shi,S.,Condon,R.G.,Deng,L.,Saunders,J.,Hung,F.,Tsao,Y.S.,Liu,Z.AHigh-throughputAutomatedPlatformfortheDevelopmentofManufacturingCellLinesforProteinTherapeutics.J.Vis.Exp.(55),e3010,doi:10.3791/3010(2011).AbstractThefast-growingbiopharmaceuticalindustrydemandsspeedydevelopmentofhighlyefficientandreliableproductionsystemstomeettheincreasingrequirementfordrugsupplies.Thegenerationofproductioncelllineshastraditionallyinvolvedmanualoperationsthatarelabor-intensive,low-throughputandvulnerabletohumanerrors.Wereporthereanintegratedhigh-throughputandautomatedplatformfordevelopmentofmanufacturingcelllinesfortheproductionofproteintherapeutics.ThecombinationofBDFACSAriaCellSorter,CloneSelectImagerandTECANFreedomEVOliquidhandlingsystemhasenabledahigh-throughputandmoreefficientcelllinedevelopmentprocess.Inthisoperation,productionhostcellsarefirsttransfectedwithanexpressionvectorcarryingthegeneofinterest1,followedbythetreatmentwithaselectionagent.Thestably-transfectedcellsarethenstainedwithfluorescence-labeledanti-humanIgGantibody,andaresubsequentlysubjecttoflowcytometryanalysis2-4.Highlyproductivecellsareselectedbasedonfluorescenceintensityandareisolatedbysingle-cellsortingonaBDFACSAria.Colonyformationfromsingle-cellstagewasdetectedmicroscopicallyandaseriesoftime-lapsdigitalimagesaretakenbyCloneSelectImagerforthedocumentationofc...
