Versatile microscale screening platform for improving recombinant protein productivity in Chinese hamster ovary cells

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dc.contributor.authorHansen, Henning Gramko
dc.contributor.authorNilsson, Claes Nymandko
dc.contributor.authorLund, Anne Mathildeko
dc.contributor.authorKol, Stefanko
dc.contributor.authorGrav, Lise Marieko
dc.contributor.authorLundqvist, Magnusko
dc.contributor.authorRockberg, Johanko
dc.contributor.authorLee, Gyun Minko
dc.contributor.authorAndersen, Mikael Rordamko
dc.contributor.authorKildegaard, Helene Faustrupko
dc.date.accessioned2016-04-20T06:49:18Z-
dc.date.available2016-04-20T06:49:18Z-
dc.date.created2016-01-04-
dc.date.created2016-01-04-
dc.date.issued2015-12-
dc.identifier.citationSCIENTIFIC REPORTS, v.5-
dc.identifier.issn2045-2322-
dc.identifier.urihttp://hdl.handle.net/10203/205505-
dc.description.abstractChinese hamster ovary (CHO) cells are widely used as cell factories for the production of biopharmaceuticals. In contrast to the highly optimized production processes for monoclonal antibody (mAb)-based biopharmaceuticals, improving productivity of non-mAb therapeutic glycoproteins is more likely to reduce production costs significantly. The aim of this study was to establish a versatile target gene screening platform for improving productivity for primarily non-mAb glycoproteins with complete interchangeability of model proteins and target genes using transient expression. The platform consists of four techniques compatible with 96-well microplates: lipid-based transient transfection, cell cultivation in microplates, cell counting and antibody-independent product titer determination based on split-GFP complementation. We were able to demonstrate growth profiles and volumetric productivity of CHO cells in 96-half-deepwell microplates comparable with those obtained in shake flasks. In addition, we demonstrate that split-GFP complementation can be used to accurately measure relative titers of therapeutic glycoproteins. Using this platform, we were able to detect target gene-specific increase in titer and specific productivity of two non-mAb glycoproteins. In conclusion, the platform provides a novel miniaturized and parallelisable solution for screening target genes and holds the potential to unravel genes that can enhance the secretory capacity of CHO cells.-
dc.languageEnglish-
dc.publisherNATURE PUBLISHING GROUP-
dc.subjectGREEN FLUORESCENT PROTEIN-
dc.subjectCHO-CELLS-
dc.subjectTRANSIENT TRANSFECTION-
dc.subjectMAMMALIAN-CELLS-
dc.subjectANTIBODY-PRODUCTION-
dc.subjectMICROTITER PLATES-
dc.subjectCURRENT STATE-
dc.subjectEXPRESSION-
dc.subjectOVEREXPRESSION-
dc.subjectINHIBITION-
dc.titleVersatile microscale screening platform for improving recombinant protein productivity in Chinese hamster ovary cells-
dc.typeArticle-
dc.identifier.wosid000366190500001-
dc.identifier.scopusid2-s2.0-84949656277-
dc.type.rimsART-
dc.citation.volume5-
dc.citation.publicationnameSCIENTIFIC REPORTS-
dc.identifier.doi10.1038/srep18016-
dc.embargo.liftdate9999-12-31-
dc.embargo.terms9999-12-31-
dc.contributor.localauthorLee, Gyun Min-
dc.contributor.nonIdAuthorHansen, Henning Gram-
dc.contributor.nonIdAuthorNilsson, Claes Nymand-
dc.contributor.nonIdAuthorLund, Anne Mathilde-
dc.contributor.nonIdAuthorKol, Stefan-
dc.contributor.nonIdAuthorGrav, Lise Marie-
dc.contributor.nonIdAuthorLundqvist, Magnus-
dc.contributor.nonIdAuthorRockberg, Johan-
dc.contributor.nonIdAuthorAndersen, Mikael Rordam-
dc.contributor.nonIdAuthorKildegaard, Helene Faustrup-
dc.description.isOpenAccessY-
dc.type.journalArticleArticle-
dc.subject.keywordPlusGREEN FLUORESCENT PROTEIN-
dc.subject.keywordPlusCHO-CELLS-
dc.subject.keywordPlusTRANSIENT TRANSFECTION-
dc.subject.keywordPlusMAMMALIAN-CELLS-
dc.subject.keywordPlusANTIBODY-PRODUCTION-
dc.subject.keywordPlusMICROTITER PLATES-
dc.subject.keywordPlusCURRENT STATE-
dc.subject.keywordPlusEXPRESSION-
dc.subject.keywordPlusOVEREXPRESSION-
dc.subject.keywordPlusINHIBITION-
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