Titer of trastuzumab produced by a Chinese hamster ovary cell line is associated with tricarboxylic acid cycle activity rather than lactate metabolism

J Biosci Bioeng. 2015 Apr;119(4):478-85. doi: 10.1016/j.jbiosc.2014.09.017. Epub 2014 Nov 7.

Abstract

Achieving high productivity and quality is the final goal of therapeutic antibody development, but the productivity and quality of antibodies are known to be substantially dependent on the nature of the cell lines expressing the antibodies. We characterized two contrasting cell lines that produce trastuzumab, namely cell line A with a high titer and a low aggregate content and cell line B with a low titer and a high aggregate content to identify the causes of the differences. We observed the following differences: cell growth (A > B), proportion of defucosylated oligosaccharides on antibodies (A < B), and proportion of covalent antibody aggregates (A > B). Our results suggest that the high monoclonal antibody (mAb) titers in cell line A is associated with the high proliferation and is not caused by the lactate metabolism shift (switching from lactate production to net lactate consumption). Rather, these differences can be accounted for by the following: levels of tricarboxylic acid cycle intermediates (A > B), ammonium ion levels (A ≤ B), and oxidative stress (A > B).

Keywords: Antibody aggregate; Antibody productivity; Antibody quality; Cell line; Chinese hamster ovary cell; Fucosylation; Lactate metabolism shift; Oxidative stress; Therapeutic antibody; Tricarboxylic acid cycle intermediates.

MeSH terms

  • Ammonium Compounds / analysis
  • Ammonium Compounds / metabolism
  • Animals
  • Antibodies, Monoclonal / analysis
  • Antibodies, Monoclonal / biosynthesis
  • Antibodies, Monoclonal / chemistry
  • Antibodies, Monoclonal, Humanized / analysis*
  • Antibodies, Monoclonal, Humanized / biosynthesis*
  • CHO Cells
  • Cell Proliferation
  • Citric Acid Cycle / physiology*
  • Cricetinae
  • Cricetulus
  • Fucose / metabolism
  • Guanosine Diphosphate / metabolism
  • Lactic Acid / metabolism*
  • Metabolome
  • Oxidative Stress
  • Trastuzumab

Substances

  • Ammonium Compounds
  • Antibodies, Monoclonal
  • Antibodies, Monoclonal, Humanized
  • Guanosine Diphosphate
  • Fucose
  • Lactic Acid
  • Trastuzumab