Journal article
BCAT1 and BCAT2 disruption in CHO cells has cell line-dependent effects
Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark1
Quantitative Modeling of Cell Metabolism, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark2
CHO Core, Translational Management, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark3
CHO Cell Line Engineering and Design, Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark4
Department of Biotechnology and Biomedicine, Technical University of Denmark5
In recombinant protein expression using Chinese hamster ovary (CHO) cells, chemically defined media contain essential amino acids such as branched chain amino acids (BCAAs) leucine, isoleucine and valine. Availability of amino acids is critical as these are building blocks for protein synthesis. However, breakdown of amino acids can lead to build up of toxic intermediates and metabolites that decrease cell growth, productivity and product quality.
BCAA catabolism also hampers the usage of BCAAs for protein synthesis. In this work we studied the effects of disrupting the genes responsible for the first step of BCAA catabolism: branched chain aminotransferase 1 (Bcat1) and branched chain aminotransferase 2 (Bcat2). We evaluated the effect of disrupting the genes individually and in combination, and examined the effects in producer and non-producer host cells.
Our experiments show that Bcat1 disruption improves cell growth in producer cells, but not in non-producers. Conversely, Bcat2 has a minor negative effect on growth in producer cells, and none in non-producers. Combined Bcat1 and Bcat2 disruption improves growth in producer cells. By-product metabolism is cell line-, clone- and producer-dependent.
Overall, our results show that the effects of targeting Bcat1 and/or Bcat2 are cell line-dependent, and seemingly linked to the burden of recombinant protein expression.
Language: | English |
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Year: | 2019 |
Pages: | 24-31 |
ISSN: | 18734863 and 01681656 |
Types: | Journal article |
DOI: | 10.1016/j.jbiotec.2019.08.017 |
ORCIDs: | Schubert, Mikkel , Andersen, Mikael Rørdam , Pereira, Sara , Ley, Daniel , Grav, Lise Marie and Kildegaard, Helene Faustrup |