Enhanced glycoprotein production in HEK-293 cells expressing pyruvate carboxylase

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DOIResolve DOI: http://doi.org/10.1016/j.ymben.2011.05.004
AuthorSearch for: ; Search for:
Journal titleMetabolic Engineering
Pages19; # of pages: 9
Subject13C-metabolic flux analysis; Ammonia; article; bio; Biotechnology; Canada; Carbon; Cells; Fed-batch; Glycoproteins; HEK-293; Hek-293 cells; HEK293; interferon; Lactate; metabolism; Pyruvate Carboxylase; Recombinant interferon; Scopus; Substrates; Yeast
AbstractThere 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-+¦2b 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 13C-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. -¬ 2011 Elsevier
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AffiliationNRC Biotechnology Research Institute; National Research Council Canada
Peer reviewedYes
NRC number52785
NPARC number18335890
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Record identifierf1b9f026-a74d-45b2-aec1-b896fdd872a7
Record created2011-07-29
Record modified2016-05-09
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