Exploiting the metabolism of PYC expressing HEK293 cells in fed-batch cultures
| Autor: | Yves Durocher, Cédric Vallée, Olivier Henry |
|---|---|
| Rok vydání: | 2014 |
| Předmět: |
Animal cell lines
phenotype Cells Clone (cell biology) Bioengineering animal cell yeast Biology Applied Microbiology and Biotechnology law.invention Automation Glucose concentration Bioreactors law Control Fed batches Batch cell culture Humans glucose Fed-batch cultures batch process cell strain HEK293 Pyruvate Carboxylase Cell growth HEK 293 cells fed batch culture Batch data processing cell clone General Medicine Metabolism Recombinant Proteins Pyruvate carboxylase enzyme metabolism HEK293 Cells Process enhancements Biochemistry Batch Cell Culture Techniques Cell culture Recombinant DNA Maximum Cell Density Cytology recombinant alpha2b interferon Recombinant yeast cell density Biotechnology |
| Zdroj: | Journal of Biotechnology. 169:63-70 |
| ISSN: | 0168-1656 |
| DOI: | 10.1016/j.jbiotec.2013.11.002 |
| Popis: | The 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. |
| Databáze: | OpenAIRE |
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