DC Field | Value | Language |
---|---|---|
dc.contributor.author | Hansen, Henning Gram | ko |
dc.contributor.author | Nilsson, Claes Nymand | ko |
dc.contributor.author | Lund, Anne Mathilde | ko |
dc.contributor.author | Kol, Stefan | ko |
dc.contributor.author | Grav, Lise Marie | ko |
dc.contributor.author | Lundqvist, Magnus | ko |
dc.contributor.author | Rockberg, Johan | ko |
dc.contributor.author | Lee, Gyun Min | ko |
dc.contributor.author | Andersen, Mikael Rordam | ko |
dc.contributor.author | Kildegaard, Helene Faustrup | ko |
dc.date.accessioned | 2016-04-20T06:49:18Z | - |
dc.date.available | 2016-04-20T06:49:18Z | - |
dc.date.created | 2016-01-04 | - |
dc.date.created | 2016-01-04 | - |
dc.date.issued | 2015-12 | - |
dc.identifier.citation | SCIENTIFIC REPORTS, v.5 | - |
dc.identifier.issn | 2045-2322 | - |
dc.identifier.uri | http://hdl.handle.net/10203/205505 | - |
dc.description.abstract | Chinese 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.language | English | - |
dc.publisher | NATURE PUBLISHING GROUP | - |
dc.subject | GREEN FLUORESCENT PROTEIN | - |
dc.subject | CHO-CELLS | - |
dc.subject | TRANSIENT TRANSFECTION | - |
dc.subject | MAMMALIAN-CELLS | - |
dc.subject | ANTIBODY-PRODUCTION | - |
dc.subject | MICROTITER PLATES | - |
dc.subject | CURRENT STATE | - |
dc.subject | EXPRESSION | - |
dc.subject | OVEREXPRESSION | - |
dc.subject | INHIBITION | - |
dc.title | Versatile microscale screening platform for improving recombinant protein productivity in Chinese hamster ovary cells | - |
dc.type | Article | - |
dc.identifier.wosid | 000366190500001 | - |
dc.identifier.scopusid | 2-s2.0-84949656277 | - |
dc.type.rims | ART | - |
dc.citation.volume | 5 | - |
dc.citation.publicationname | SCIENTIFIC REPORTS | - |
dc.identifier.doi | 10.1038/srep18016 | - |
dc.embargo.liftdate | 9999-12-31 | - |
dc.embargo.terms | 9999-12-31 | - |
dc.contributor.localauthor | Lee, Gyun Min | - |
dc.contributor.nonIdAuthor | Hansen, Henning Gram | - |
dc.contributor.nonIdAuthor | Nilsson, Claes Nymand | - |
dc.contributor.nonIdAuthor | Lund, Anne Mathilde | - |
dc.contributor.nonIdAuthor | Kol, Stefan | - |
dc.contributor.nonIdAuthor | Grav, Lise Marie | - |
dc.contributor.nonIdAuthor | Lundqvist, Magnus | - |
dc.contributor.nonIdAuthor | Rockberg, Johan | - |
dc.contributor.nonIdAuthor | Andersen, Mikael Rordam | - |
dc.contributor.nonIdAuthor | Kildegaard, Helene Faustrup | - |
dc.description.isOpenAccess | Y | - |
dc.type.journalArticle | Article | - |
dc.subject.keywordPlus | GREEN FLUORESCENT PROTEIN | - |
dc.subject.keywordPlus | CHO-CELLS | - |
dc.subject.keywordPlus | TRANSIENT TRANSFECTION | - |
dc.subject.keywordPlus | MAMMALIAN-CELLS | - |
dc.subject.keywordPlus | ANTIBODY-PRODUCTION | - |
dc.subject.keywordPlus | MICROTITER PLATES | - |
dc.subject.keywordPlus | CURRENT STATE | - |
dc.subject.keywordPlus | EXPRESSION | - |
dc.subject.keywordPlus | OVEREXPRESSION | - |
dc.subject.keywordPlus | INHIBITION | - |
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