Versatile Microscale Screening Platform for Improving Recombinant Protein Productivity in Chinese Hamster Ovary Cells

Sci Rep. 2015 Dec 11;5:18016. doi: 10.1038/srep18016.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 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 / biosynthesis*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics*
  • Reproducibility of Results

Substances

  • Recombinant Fusion Proteins
  • Recombinant Proteins