Essential Site Maintenance: Authorea-powered sites will be updated circa 15:00-17:00 Eastern on Tuesday 5 November.
There should be no interruption to normal services, but please contact us at [email protected] in case you face any issues.

loading page

Metabolic Analysis of the Asparagine and Glutamine Dynamics in an Industrial CHO Fed-Batch Process
  • +7
  • Brian Kirsch,
  • Sandra Bennun,
  • Adam Mendez,
  • Amy Johnson,
  • Hongxia Wang,
  • Haibo Qiu,
  • Ning Li,
  • Shawn Lawrence,
  • Hanne Bak,
  • Michael Betenbaugh
Brian Kirsch
Johns Hopkins University

Corresponding Author:[email protected]

Author Profile
Sandra Bennun
Regeneron Pharmaceuticals Inc
Author Profile
Adam Mendez
Johns Hopkins University
Author Profile
Amy Johnson
Regeneron Pharmaceuticals Inc
Author Profile
Hongxia Wang
Regeneron Pharmaceuticals Inc
Author Profile
Haibo Qiu
Regeneron Pharmaceuticals Inc
Author Profile
Ning Li
Regeneron Pharmaceuticals Inc
Author Profile
Shawn Lawrence
Regeneron Pharmaceuticals Inc
Author Profile
Hanne Bak
Regeneron Pharmaceuticals Inc
Author Profile
Michael Betenbaugh
Johns Hopkins University
Author Profile

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 13C-glucose, 13C-glutamine, and 13C-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 (NAG) 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 13C studies demonstrate that metabolic flux is process time dependent and can be modulated by varying feed composition.
12 Jan 2021Submitted to Biotechnology and Bioengineering
12 Jan 2021Submission Checks Completed
12 Jan 2021Assigned to Editor
25 Jan 2021Reviewer(s) Assigned
20 Feb 2021Editorial Decision: Revise Major
20 Feb 2021Review(s) Completed, Editorial Evaluation Pending
23 Aug 20211st Revision Received
24 Aug 2021Submission Checks Completed
24 Aug 2021Assigned to Editor
30 Aug 2021Reviewer(s) Assigned
12 Sep 2021Review(s) Completed, Editorial Evaluation Pending
12 Sep 2021Editorial Decision: Revise Major
28 Sep 20212nd Revision Received
28 Sep 2021Submission Checks Completed
28 Sep 2021Assigned to Editor
04 Oct 2021Review(s) Completed, Editorial Evaluation Pending
04 Oct 2021Editorial Decision: Accept
17 Nov 2021Published in Biotechnology and Bioengineering. 10.1002/bit.27993