BioChem - DOS

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

Scientific reports

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

<P>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.</P>

2015

Nature Publishing Group

cc-by

2045-2322

5

pp.18016

10.1038/srep18016

http://click.ndsl.kr/servlet/OpenAPIDetailView?keyValue=05787966&target=NART&cn=NART74676667

https://www.nature.com/articles/srep18016.pdf