Exploiting the metabolism of PYC expressing HEK293 cells in fed-batch cultures

  1. Get@NRC: Exploiting the metabolism of PYC expressing HEK293 cells in fed-batch cultures (Opens in a new window)
DOIResolve DOI: http://doi.org/10.1016/j.jbiotec.2013.11.002
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Journal titleJournal of Biotechnology
Pages6370; # of pages: 8
SubjectAnimal cell lines; Fed batches; Fed-batch cultures; Glucose concentration; HEK293 cells; Process enhancements; Pyruvate carboxylase; Recombinant yeast; Automation; Batch data processing; Cells; Control; Cytology; Recombinant proteins; Batch cell culture; glucose; pyruvate carboxylase; recombinant alpha2b interferon; animal cell; article; batch process; cell clone; cell density; cell strain HEK293; controlled study; enzyme metabolism; fed batch culture; nonhuman; phenotype; priority journal; yeast
AbstractThe expression of recombinant yeast pyruvate carboxylase (PYC) in animal cell lines was shown in previous studies to reduce significantly the formation of waste metabolites, although it has translated into mixed results in terms of improved cellular growth and productivity. In this work, we demonstrate that the unique phenotype of PYC expressing cells can be exploited through the application of a dynamic fed-batch strategy and lead to significant process enhancements. Metabolically engineered HEK293 cells stably producing human recombinant IFNα2b and expressing the PYC enzyme were cultured in batch and fed-batch modes. Compared to parental cells, the maximum cell density in batch was increased 1.5-fold and the culture duration was extended by 2.5 days, but the product yield was only marginally increased. Further improvements were achieved by developing and implementing a dynamic fed-batch strategy using a concentrated feed solution. The feeding was based on an automatic control-loop to maintain a constant glucose concentration. This strategy led to a further 2-fold increase in maximum cell density (up to 10.7×106cells/ml) and a final product titer of 160mg/l, representing nearly a 3-fold yield increase compared to the batch process with the parental cell clone. © 2013.
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AffiliationNational Research Council Canada (NRC-CNRC); Human Health Therapeutics (HHT-TSH)
Peer reviewedYes
NPARC number21270818
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Record identifierdf3d9042-89f0-4391-993b-3e054f2788d8
Record created2014-02-17
Record modified2016-05-09
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