title: Metabolite profiling of CHO cells: Molecular reflections of bioprocessing effectiveness.
abstract: Whilst development of medium and feeds has provided major advances in recombinant protein production in CHO cells, the fundamental understanding is limited. We have applied metabolite profiling with established robust (GC-MS) analytics to define the molecular loci by which two yield-enhancing feeds improve recombinant antibody yields from a model GS-CHO cell line. With data across core metabolic pathways, that report on metabolism within several cellular compartments, these data identify key metabolites and events associated with increased cell survival and specific productivity of cells. Of particular importance, increased process efficiency was linked to the functional activity of the mitochondria, with the amount and time course of use/production of intermediates of the citric acid cycle, for uses such as lipid biosynthesis, precursor generation and energy production, providing direct indicators of cellular status with respect to productivity. The data provide clear association between specific cellular metabolic indicators and cell process efficiency, extending from prior indications of the relevance of lactate metabolic balance to other redox sinks (glycerol, sorbitol and threitol). The information, and its interpretation, identifies targets for engineering cell culture efficiency, either from genetic or environmental perspectives, and greater understanding of the significance of specific medium components towards overall CHO cell bioprocessing.
authors.affiliation: Faculty of Life Sciences, The University of Manchester, Manchester, UK.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Sellick
authors.affiliation: Faculty of Life Sciences, The University of Manchester, Manchester, UK.
authors.forename: Alexandra S
authors.initials: AS
authors.lastname: Croxford
authors.affiliation: Faculty of Life Sciences, The University of Manchester, Manchester, UK.
authors.forename: Arfa R
authors.initials: AR
authors.lastname: Maqsood
authors.affiliation: The School of Chemical Engineering and Analytical Sciences, Manchester Institute of Biotechnology, The University of -Manchester, Manchester, UK.
authors.forename: Gill M
authors.initials: GM
authors.lastname: Stephens
authors.affiliation: The Manchester Centre for Integrative Systems Biology, Manchester Institute of Biotechnology, The University of Manchester, Manchester, UK.|Netherlands Institute for Systems Biology, VUA and UVA, Amsterdam, The Netherlands.
authors.forename: Hans V
authors.initials: HV
authors.lastname: Westerhoff
authors.affiliation: The Manchester Centre for Integrative Systems Biology, Manchester Institute of Biotechnology, The University of Manchester, Manchester, UK.|The School of Chemistry, Manchester Institute of Biotechnology, The University of Manchester, Manchester, UK.
authors.forename: Royston
authors.initials: R
authors.lastname: Goodacre
authors.affiliation: Faculty of Life Sciences, The University of Manchester, Manchester, UK. alan.dickson@manchester.ac.uk.
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Dickson
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D042541:Intracellular Space / Q000378:metabolism; D055432:Metabolomics / Q000379:methods*; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D003470:Culture Media; D011994:Recombinant Proteins
keywords: -Recombinant antibody; Bioprocessing; Gas chromatography-mass spectrometry (GC-MS); Glutamine synthetase; Metabolomics