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Analyzing clonal variation of monoclonal antibody-producing CHO cell lines using an in silico metabolomic platform

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Ghorbaniaghdam, Atefeh, Chen, Jingkui, Henry, Olivier, Jolicoeur, Mario·2014·PLoS ONE
DOI 10.1371/journal.pone.0090832PMC PMC3954614
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Abstract

Monoclonal antibody producing Chinese hamster ovary (CHO) cells have been shown to undergo metabolic changes when engineered to produce high titers of recombinant proteins. In this work, we have studied the distinct metabolism of CHO cell clones harboring an efficient inducible expression system, based on the cumate gene switch, and displaying different expression levels, high and low productivities, compared to that of the parental cells from which they were derived. A kinetic model for CHO cell metabolism was further developed to include metabolic regulation. Model calibration was performed using intracellular and extracellular metabolite profiles obtained from shake flask batch cultures. Model simulations of intracellular fluxes and ratios known as biomarkers revealed significant changes correlated with clonal variation but not to the recombinant protein expression level. Metabolic flux distribution mostly differs in the reactions involving pyruvate metabolism, with an increased net flux of pyruvate into the tricarboxylic acid (TCA) cycle in the high-producer clone, either being induced or non-induced with cumate. More specifically, CHO cell metabolism in this clone was characterized by an efficient utilization of glucose and a high pyruvate dehydrogenase flux. Moreover, the high-producer clone shows a high rate of anaplerosis from pyruvate to oxaloacetate, through pyruvate carboxylase and from glutamate to α-ketoglutarate, through glutamate dehydrogenase, and a reduced rate of cataplerosis from malate to pyruvate, through malic enzyme. Indeed, the increase of flux through pyruvate carboxylase was not driven by an increased anabolic demand. It is in fact linked to an increase of the TCA cycle global flux, which allows better regulation of higher redox and more efficient metabolic states. To the best of our knowledge, this is the first time a dynamic in silico platform is proposed to analyze and compare the metabolomic behavior of different CHO clones.

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  • A bottleneck at the malate dehydrogenase II (MDHII) level was characterized for the tricarboxylic acid (TCA) cycle in CHO cells and pyruvate metabolism was shown to vary between high producing and low producing anti-CD20 CHO clones.

    “A bottleneck at the malate dehydrogenase II (MDHII) level was characterized for the tricarboxylic acid (TCA) cycle in CHO cells and pyruvate metabolism was shown to vary between high producing and low producing anti-CD20 CHO clones (Chong et al., 2010;Ghorbaniaghdam et al., 2014).”

    From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Ghorbaniaghdam et al., 2014

    The Materials and Methods section identifies the high- and low-producing clones as producers of anti-CD20 antibody. The Discussion reports different pyruvate dehydrogenase and pyruvate carboxylase fluxes between those clones, supporting the pyruvate-metabolism part of the sentence. The passages from this paper do not characterize a TCA-cycle bottleneck at MDHII. (LLM, conf=0.88)

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Authors
Ghorbaniaghdam, A., Chen, J., Henry, O., & Jolicoeur, M.
Affiliations
Polytechnique Montréal (CA)
ORCID
Henry, Olivier
Venue
PLoS ONE
Publisher
Public Library of Science (PLoS)
Published
2014-03-14
Volume
9
Issue
3
Pages
e90832
ISSN
1932-6203 (ISSN-L)
DOI
10.1371/journal.pone.0090832
PMC
PMC3954614
OpenAlex
W2001064853
Cited by
71
References
2
Citations by year
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FWCI
3.42
Type
journal article
Language
en
Update notices
correction (2014-08-01)
Open access
Licence
Locations
  • PLoS ONE (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PolyPublie (École Polytechnique de Montréal) (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • Figshare (submittedVersion, cc-by)
  • PolyPublie (École Polytechnique de Montréal) (submittedVersion)
Funders
Natural Sciences and Engineering Research Council of Canada
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Keywords
monoclonal antibody production, clonal variation, inducible expression system, metabolic flux distribution, TCA cycle, pyruvate metabolism, anaplerosis, cataplerosis, pyruvate carboxylase, glutamate dehydrogenase, malic enzyme, metabolic regulation, kinetic metabolic models, recombinant protein productivity, metabolomic biomarkers
MeSH terms
Animals, Antibodies, Monoclonal, Citric Acid Cycle, Cricetulus, Ketoglutaric Acids, Models, Theoretical, Pyruvate Decarboxylase, CHO Cells, Pyruvic Acid, Metabolomics

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