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Cited by 39Viral Infectious Diseases and Gene Expression in InsectsMicrobial Metabolic Engineering and BioproductionCancer, Hypoxia, and Metabolism

Metabolic analysis of the asparagine and glutamine dynamics in an industrial Chinese hamster ovary fed-batch process

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Kirsch, Brian J., Bennun, Sandra V., Mendez, Adam, Johnson, Amy S., Wang, Hongxia, Qiu, Haibo, Li, Ning, Lawrence, Shawn M. +2 more·2022·Biotechnology and Bioengineering
DOI 10.1002/bit.27993PMC PMC9305493

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Abstract

Chinese hamster ovary (CHO) cell lines are grown in cultures with varying asparagine and glutamine concentrations, but further study is needed to characterize the interplay between these amino acids. By following 13 C‐glucose, 13 C‐glutamine, and 13 C‐asparagine tracers using metabolic flux analysis (MFA), CHO cell metabolism was characterized in an industrially relevant fed‐batch process under glutamine supplemented and low glutamine conditions during early and late exponential growth. For both conditions MFA revealed glucose as the primary carbon source to the tricarboxylic acid (TCA) cycle followed by glutamine and asparagine as secondary sources. Early exponential phase CHO cells prefer glutamine over asparagine to support the TCA cycle under the glutamine supplemented condition, while asparagine was critical for TCA activity for the low glutamine condition. Overall TCA fluxes were similar for both conditions due to the trade‐offs associated with reliance on glutamine and/or asparagine. However, glutamine supplementation increased fluxes to alanine, lactate and enrichment of glutathione, N ‐acetyl‐glucosamine and pyrimidine‐containing‐molecules. The late exponential phase exhibited reduced central carbon metabolism dominated by glucose, while lactate reincorporation and aspartate uptake were preferred over glutamine and asparagine. These 13 C studies demonstrate that metabolic flux is process time dependent and can be modulated by varying feed composition.

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  • Asparagine plays a central metabolic role in mammalian cell culture, acting as a nitrogen donor for amino acid and nucleotide biosynthesis, supporting protein translation, and contributing to TCA cycle intermediates that sustain biomass growth and productivity.

    “Asparagine plays a central metabolic role in mammalian cell culture, acting as a nitrogen donor for amino acid and nucleotide biosynthesis, supporting protein translation, and contributing to TCA cycle intermediates that sustain biomass growth and productivity [48].”

    From Soft sensing of intracellular states for CHO cell bioprocessing with Ensemble Kalman Filters (2026) · cites this paper as [48]

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Cited by

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Authors
Kirsch, B. J., Bennun, S. V., Mendez, A., Johnson, A. S., Wang, H., Qiu, H., Li, N., Lawrence, S. M., Bak, H., & Betenbaugh, M. J.
Affiliations
  • Johns Hopkins University (US)
  • Regeneron (United States) (US)
Corresponding author
Bennun, Sandra V.
ORCID
Venue
Biotechnology and Bioengineering
Publisher
Wiley
Published
2022-01-06
Volume
119
Issue
3
Pages
807-819
ISSN
  • 0006-3592 (ISSN-L)
  • 1097-0290
DOI
10.1002/bit.27993
PMC
PMC9305493
OpenAlex
W3214722919
Cited by
39
References
5
Citations by year
2026: 11 · 2025: 10 · 2024: 9 · 2023: 6 · 2022: 2 · 2021: 1
FWCI
1.41
Type
journal article
Language
en
Open access
Licence
Locations
  • Biotechnology and Bioengineering (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
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 › Cancer Research › Cancer, Hypoxia, and Metabolism
Keywords
CHO cells, asparagine, glutamine, fed-batch fermentation, metabolic flux analysis, TCA cycle, carbon source utilization, glutamine supplementation, 13C tracer, lactate metabolism, glutathione, N-acetylglucosamine, pyrimidine metabolism, exponential growth phase, cellular metabolism
MeSH terms
Animals, Asparagine, Cricetulus, Glucose, Glutamine, Cricetinae, CHO Cells, Lactic Acid

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