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

• Published in: Scientific reports 2015-12, Vol.5 (1), p.18016, Article 18016
• Main Authors: Hansen, Henning Gram, Nilsson, Claes Nymand, Lund, Anne Mathilde, Kol, Stefan, Grav, Lise Marie, Lundqvist, Magnus, Rockberg, Johan, Lee, Gyun Min, Andersen, Mikael Rørdam, Kildegaard, Helene Faustrup
• Format: Article
• Language: English
• Subjects: 13/44; 14/63; 38/35; 42/34; 49/109; 49/47; 631/1647; 631/61; 82/80; Animals; Cell culture; Cell Culture Techniques; CHO Cells; Complementation; Cricetinae; Cricetulus; Enzyme-Linked Immunosorbent Assay; Gene Expression; Genes; Genes, Reporter; Glycoproteins; Glycosylation; High-Throughput Screening Assays; Humanities and Social Sciences; Monoclonal antibodies; multidisciplinary; Production costs; Productivity; Protein Interaction Domains and Motifs; Recombinant Fusion Proteins; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Recombinant Proteins - genetics; Reproducibility of Results; Science; Toxicity; Transfection
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep18016

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.