Back to Search
Start Over
Enhanced glycoprotein production in HEK-293 cells expressing pyruvate carboxylase.
- Source :
-
Metabolic engineering [Metab Eng] 2011 Sep; Vol. 13 (5), pp. 499-507. Date of Electronic Publication: 2011 May 23. - Publication Year :
- 2011
-
Abstract
- There is an imperative need for expression systems allowing the efficient and robust manufacturing of high quality glycoproteins. In the present work, HEK-293 cells stably expressing interferon-α2b were further engineered with the insertion of the yeast pyruvate carboxylase 2 gene. In batch cultures, marked reductions in lactate and ammonia production were observed compared to the parental cell clone. Although the maximum specific growth rate remained unchanged, the altered metabolism led to a 2-fold increase in maximum cell density and 33% increase in the integral of viable cell concentration and interferon production yield. The underlying metabolic changes were further investigated using various (13)C-labeled substrates and measuring the resulting lactate mass isotopomer distributions. Simultaneous metabolite and isotopomer balancing allowed the accurate determination of key intracellular fluxes. Such detailed and quantitative knowledge about the central carbon metabolism of the cells is instrumental to further support the development of high-yield fed-batch processes.<br /> (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Subjects :
- Cell Survival
Glycoproteins genetics
HEK293 Cells
Humans
Interferon alpha-2
Interferon-alpha genetics
Pyruvate Carboxylase genetics
Recombinant Proteins
Saccharomyces cerevisiae genetics
Saccharomyces cerevisiae Proteins genetics
Glycoproteins biosynthesis
Interferon-alpha biosynthesis
Pyruvate Carboxylase biosynthesis
Saccharomyces cerevisiae enzymology
Saccharomyces cerevisiae Proteins biosynthesis
Subjects
Details
- Language :
- English
- ISSN :
- 1096-7184
- Volume :
- 13
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Metabolic engineering
- Publication Type :
- Academic Journal
- Accession number :
- 21624497
- Full Text :
- https://doi.org/10.1016/j.ymben.2011.05.004