title: Application of13C flux analysis to identify high-productivity CHO metabolic phenotypes.
abstract: Industrial bioprocesses place high demands on the energy metabolism of host cells to meet biosynthetic requirements for maximal protein expression. Identifying metabolic phenotypes that promote high expression is therefore a major goal of the biotech industry. We conducted a series of13C flux analysis studies to examine the metabolic response to IgG expression during early stationary phase of CHO cell cultures grown in 3L fed-batch bioreactors. We examined eight clones expressing four different IgGs and compared with three non-expressing host-cell controls. Some clones were genetically manipulated to be apoptosis-resistant by expressing Bcl-2Δ, which correlated with increased IgG production and elevated glucose metabolism. The metabolic phenotypes of the non-expressing, IgG-expressing, and Bcl-2Δ/IgG-expressing clones were fully segregated by hierarchical clustering analysis. Lactate consumption and citric acid cycle fluxes were most strongly associated with specific IgG productivity. These studies indicate that enhanced oxidative metabolism is a characteristic of high-producing CHO cell lines.
authors.affiliation: Department of Chemical and Biomolecular Engineering, Vanderbilt University, PMB 351604, Nashville, TN 37235-1604, USA.
authors.forename: Neil
authors.initials: N
authors.lastname: Templeton
authors.affiliation: Pharmaceutical Development and Manufacturing Sciences, Janssen Research & Development, 1400 McKean Road, Spring House, PA 19477, USA.
authors.forename: Kevin D
authors.initials: KD
authors.lastname: Smith
authors.affiliation: Department of Chemical and Biomolecular Engineering, Vanderbilt University, PMB 351604, Nashville, TN 37235-1604, USA.
authors.forename: Allison G
authors.initials: AG
authors.lastname: McAtee-Pereira
authors.affiliation: Pharmaceutical Development and Manufacturing Sciences, Janssen Research & Development, 1400 McKean Road, Spring House, PA 19477, USA.
authors.forename: Haimanti
authors.initials: H
authors.lastname: Dorai
authors.affiliation: Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 North Charles Street, Baltimore, MD 21218, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Betenbaugh
authors.affiliation: Pharmaceutical Development and Manufacturing Sciences, Janssen Research & Development, 1400 McKean Road, Spring House, PA 19477, USA.
authors.forename: Steven E
authors.initials: SE
authors.lastname: Lang
authors.affiliation: Department of Chemical and Biomolecular Engineering, Vanderbilt University, PMB 351604, Nashville, TN 37235-1604, USA; Department of Molecular Physiology and Biophysics, Vanderbilt University PMB 351604, Nashville, TN 37235-1604, USA. Electronic address: j.d.young@vanderbilt.edu.
authors.forename: Jamey D
authors.initials: JD
authors.lastname: Young
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics; D016466:CHO Cells; D002247:Carbon Isotopes / Q000737:chemistry*; D002952:Citric Acid Cycle; D003412:Cricetulus; D015870:Gene Expression; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000235:genetics; D007553:Isotope Labeling; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D002247:Carbon Isotopes; D007074:Immunoglobulin G; D011994:Recombinant Proteins
keywords: Chinese hamster ovary; Citric acid cycle; IgG expression; Lactate shift; Metabolic flux analysis