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

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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.
Publisher
NATURE PUBLISHING GROUP
Issue Date
2015-12
Language
English
Article Type
Article
Keywords

GREEN FLUORESCENT PROTEIN; CHO-CELLS; TRANSIENT TRANSFECTION; MAMMALIAN-CELLS; ANTIBODY-PRODUCTION; MICROTITER PLATES; CURRENT STATE; EXPRESSION; OVEREXPRESSION; INHIBITION

Citation

SCIENTIFIC REPORTS, v.5

ISSN
2045-2322
DOI
10.1038/srep18016
URI
http://hdl.handle.net/10203/205505
Appears in Collection
BS-Journal Papers(저널논문)
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