Journal article
Versatile microscale screening platform for improving recombinant protein productivity in Chinese hamster ovary cells
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
CHO Cell Line Engineering and Design, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
Department of Systems Biology, Technical University of Denmark3
Network Engineering of Eukaryotic Cell Factories, Department of Systems Biology, Technical University of Denmark4
CHO Core, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark5
High Throughput Molecular Bioscience, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark6
KTH Royal Institute of Technology7
Korea Advanced Institute of Science and Technology8
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.
Language: | English |
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Publisher: | Nature Publishing Group |
Year: | 2015 |
Pages: | 18016 |
ISSN: | 20452322 |
Types: | Journal article |
DOI: | 10.1038/srep18016 |
ORCIDs: | Hansen, Henning Gram , Kol, Stefan , Grav, Lise Marie , Andersen, Mikael Rørdam and Kildegaard, Helene Faustrup |
Animals CHO Cells Cell Culture Techniques Cricetinae Cricetulus Enzyme-Linked Immunosorbent Assay Gene Expression Genes, Reporter Glycosylation High-Throughput Screening Assays Protein Interaction Domains and Motifs Recombinant Fusion Proteins Recombinant Proteins Reproducibility of Results