title: Combined metabolomics and proteomics reveals hypoxia as a cause of lower productivity on scale-up to a 5000-liter CHO bioprocess.
abstract: Large-scale bioprocessing is key to the successful manufacturing of a biopharmaceutical. However, cell viability and productivity are often lower in the scale-up from laboratory to production. In this study, we analyzed CHO cells, which showed lower percent viabilities and productivity in a 5-KL production scale bioreactor compared to a 20-L bench-top scale under seemingly identical process parameters. An increase in copper concentration in the media from 0.02 µM to 0.4 µM led to a doubling of percent viability in the production scale albeit still at a lower level than the bench-top scale. Combined metabolomics and proteomics revealed the increased copper reduced the presence of reactive oxygen species (ROS) in the 5-KL scale process. The reduction in oxidative stress was supported by the increased level of glutathione peroxidase in the lower copper level condition. The excess ROS was shown to be due to hypoxia (intermittent), as evidenced by the reduction in fibronectin with increased copper. The 20-L scale showed much less hypoxia and thus less excess ROS generation, resulting in little to no impact to productivity with the increased copper in the media. The study illustrates the power of 'Omics in aiding in the understanding of biological processes in biopharmaceutical production.
authors.affiliation: Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, USA.
authors.forename: Yuanwei
authors.initials: Y
authors.lastname: Gao
authors.affiliation: Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, USA.
authors.forename: Somak
authors.initials: S
authors.lastname: Ray
authors.affiliation: Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, USA.
authors.forename: Shujia
authors.initials: S
authors.lastname: Dai
authors.affiliation: Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, USA.
authors.forename: Alexander R
authors.initials: AR
authors.lastname: Ivanov
authors.affiliation: Biologics Development, Global Manufacturing and Supply, Bristol-Myers Squibb, 38 Jackson Road, Devens, MA, USA.
authors.forename: Nicholas R
authors.initials: NR
authors.lastname: Abu-Absi
authors.affiliation: Biologics Development, Global Manufacturing and Supply, Bristol-Myers Squibb, 38 Jackson Road, Devens, MA, USA.
authors.forename: Amanda M
authors.initials: AM
authors.lastname: Lewis
authors.affiliation: Biologics Development, Global Manufacturing and Supply, Bristol-Myers Squibb, 38 Jackson Road, Devens, MA, USA.
authors.forename: Zhuangrong
authors.initials: Z
authors.lastname: Huang
authors.affiliation: Biologics Development, Global Manufacturing and Supply, Bristol-Myers Squibb, 38 Jackson Road, Devens, MA, USA.
authors.forename: Zizhuo
authors.initials: Z
authors.lastname: Xing
authors.affiliation: Biologics Development, Global Manufacturing and Supply, Bristol-Myers Squibb, 38 Jackson Road, Devens, MA, USA.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Borys
authors.affiliation: Biologics Development, Global Manufacturing and Supply, Bristol-Myers Squibb, 38 Jackson Road, Devens, MA, USA.
authors.forename: Zheng Jian
authors.initials: ZJ
authors.lastname: Li
authors.affiliation: Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA, USA. b.karger@neu.edu.
authors.forename: Barry L
authors.initials: BL
authors.lastname: Karger
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques / Q000379:methods*; D019149:Bioreactors; D016466:CHO Cells; D015687:Cell Hypoxia; D049109:Cell Proliferation; D002470:Cell Survival; D003300:Copper; D003412:Cricetulus; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D055432:Metabolomics / Q000379:methods*; D040901:Proteomics / Q000379:methods*; D017382:Reactive Oxygen Species / Q000378:metabolism*
chemical_list: D005353:Fibronectins; D017382:Reactive Oxygen Species; D003300:Copper
keywords: CHO cells; Copper; Hypoxia; Omics; Production scale