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Versatile microscal...
Versatile microscale screening platform for improving recombinant protein productivity in Chinese hamster ovary cells
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Hansen, Henning Gram (författare)
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Nilsson, Claes Nymand (författare)
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Lund, Anne Mathilde (författare)
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Kol, Stefan (författare)
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Grav, Lise Marie (författare)
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- Lundqvist, Magnus (författare)
- KTH,Proteomik och nanobioteknologi
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- Rockberg, Johan (författare)
- KTH,Proteomik och nanobioteknologi
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Lee, Gyun Min (författare)
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Andersen, Mikael Rordam (författare)
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Kildegaard, Helene Faustrup (författare)
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(creator_code:org_t)
- 2015-12-11
- 2015
- Engelska.
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Ingår i: Scientific Reports. - : Nature Publishing Group. - 2045-2322. ; 5
- Relaterad länk:
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https://www.nature.c...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- 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.
Ämnesord
- TEKNIK OCH TEKNOLOGIER -- Industriell bioteknik (hsv//swe)
- ENGINEERING AND TECHNOLOGY -- Industrial Biotechnology (hsv//eng)
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Hansen, Henning ...
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Nilsson, Claes N ...
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Lund, Anne Mathi ...
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Kol, Stefan
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Grav, Lise Marie
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Lundqvist, Magnu ...
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visa fler...
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Rockberg, Johan
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Lee, Gyun Min
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Andersen, Mikael ...
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Kildegaard, Hele ...
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visa färre...
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Kungliga Tekniska Högskolan