Application of a genome-scale model in tandem with enzyme assays for identification of metabolic signatures of high and low CHO cell producers

Cyrielle Calmels, Solène Arnoult, Bassem Ben Yahia, Laetitia Malphettes, Mikael Rørdam Andersen*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

Biopharmaceutical industrial processes are based on high yielding stable recombinant Chinese Hamster Ovary (CHO) cells that express monoclonal antibodies. However, the process and feeding regimes need to be adapted for each new cell line, as they all have a slightly different metabolism and product performance. A main limitation for accelerating process development is that the metabolic pathways underlying this physiological variability are not yet fully understood. This study describes the evolution of intracellular fluxes during the process for 4 industrial cell lines, 2 high producers and 2 low producers (n = 3), all of them producing a different antibody. In order to understand from a metabolic point of view the phenotypic differences observed, and to find potential targets for improving specific productivity of low producers, the analysis was supported by a tailored genome-scale model and was validated with enzymatic assays performed at different days of the process. A total of 59 reactions were examined from different key pathways, namely glycolysis, pentose phosphate pathway, TCA cycle, lipid metabolism, and oxidative phosphorylation. The intracellular fluxes did not show a metabolic correlation between high producers, but the degree of similitude observed between cell lines could be confirmed with additional experimental observations. The whole analysis led to a better understanding of the metabolic requirements for all the cell lines, allowed to the identification of metabolic bottlenecks and suggested targets for further cell line engineering. This study is a successful application of a curated genome-scale model to multiple industrial cell lines, which makes the metabolic model suitable for process platform.
Original languageEnglish
Article numbere00097
JournalMetabolic Engineering Communications
Volume9
Number of pages11
ISSN2214-0301
DOIs
Publication statusPublished - 2019

Keywords

  • Genome-scale metabolic model
  • Chinese hamster ovary
  • Flux distribution
  • Mathematical modeling
  • Metabolic engineering

Cite this

@article{07c1b78f8e1b42648b5ad74f261ebab9,
title = "Application of a genome-scale model in tandem with enzyme assays for identification of metabolic signatures of high and low CHO cell producers",
abstract = "Biopharmaceutical industrial processes are based on high yielding stable recombinant Chinese Hamster Ovary (CHO) cells that express monoclonal antibodies. However, the process and feeding regimes need to be adapted for each new cell line, as they all have a slightly different metabolism and product performance. A main limitation for accelerating process development is that the metabolic pathways underlying this physiological variability are not yet fully understood. This study describes the evolution of intracellular fluxes during the process for 4 industrial cell lines, 2 high producers and 2 low producers (n = 3), all of them producing a different antibody. In order to understand from a metabolic point of view the phenotypic differences observed, and to find potential targets for improving specific productivity of low producers, the analysis was supported by a tailored genome-scale model and was validated with enzymatic assays performed at different days of the process. A total of 59 reactions were examined from different key pathways, namely glycolysis, pentose phosphate pathway, TCA cycle, lipid metabolism, and oxidative phosphorylation. The intracellular fluxes did not show a metabolic correlation between high producers, but the degree of similitude observed between cell lines could be confirmed with additional experimental observations. The whole analysis led to a better understanding of the metabolic requirements for all the cell lines, allowed to the identification of metabolic bottlenecks and suggested targets for further cell line engineering. This study is a successful application of a curated genome-scale model to multiple industrial cell lines, which makes the metabolic model suitable for process platform.",
keywords = "Genome-scale metabolic model, Chinese hamster ovary, Flux distribution, Mathematical modeling, Metabolic engineering",
author = "Cyrielle Calmels and Sol{\`e}ne Arnoult and {Ben Yahia}, Bassem and Laetitia Malphettes and Andersen, {Mikael R{\o}rdam}",
year = "2019",
doi = "10.1016/j.mec.2019.e00097",
language = "English",
volume = "9",
journal = "Metabolic Engineering Communications",
issn = "2214-0301",
publisher = "Elsevier",

}

Application of a genome-scale model in tandem with enzyme assays for identification of metabolic signatures of high and low CHO cell producers. / Calmels, Cyrielle; Arnoult, Solène; Ben Yahia, Bassem; Malphettes, Laetitia; Andersen, Mikael Rørdam.

In: Metabolic Engineering Communications, Vol. 9, e00097, 2019.

Research output: Contribution to journalJournal articleResearchpeer-review

TY - JOUR

T1 - Application of a genome-scale model in tandem with enzyme assays for identification of metabolic signatures of high and low CHO cell producers

AU - Calmels, Cyrielle

AU - Arnoult, Solène

AU - Ben Yahia, Bassem

AU - Malphettes, Laetitia

AU - Andersen, Mikael Rørdam

PY - 2019

Y1 - 2019

N2 - Biopharmaceutical industrial processes are based on high yielding stable recombinant Chinese Hamster Ovary (CHO) cells that express monoclonal antibodies. However, the process and feeding regimes need to be adapted for each new cell line, as they all have a slightly different metabolism and product performance. A main limitation for accelerating process development is that the metabolic pathways underlying this physiological variability are not yet fully understood. This study describes the evolution of intracellular fluxes during the process for 4 industrial cell lines, 2 high producers and 2 low producers (n = 3), all of them producing a different antibody. In order to understand from a metabolic point of view the phenotypic differences observed, and to find potential targets for improving specific productivity of low producers, the analysis was supported by a tailored genome-scale model and was validated with enzymatic assays performed at different days of the process. A total of 59 reactions were examined from different key pathways, namely glycolysis, pentose phosphate pathway, TCA cycle, lipid metabolism, and oxidative phosphorylation. The intracellular fluxes did not show a metabolic correlation between high producers, but the degree of similitude observed between cell lines could be confirmed with additional experimental observations. The whole analysis led to a better understanding of the metabolic requirements for all the cell lines, allowed to the identification of metabolic bottlenecks and suggested targets for further cell line engineering. This study is a successful application of a curated genome-scale model to multiple industrial cell lines, which makes the metabolic model suitable for process platform.

AB - Biopharmaceutical industrial processes are based on high yielding stable recombinant Chinese Hamster Ovary (CHO) cells that express monoclonal antibodies. However, the process and feeding regimes need to be adapted for each new cell line, as they all have a slightly different metabolism and product performance. A main limitation for accelerating process development is that the metabolic pathways underlying this physiological variability are not yet fully understood. This study describes the evolution of intracellular fluxes during the process for 4 industrial cell lines, 2 high producers and 2 low producers (n = 3), all of them producing a different antibody. In order to understand from a metabolic point of view the phenotypic differences observed, and to find potential targets for improving specific productivity of low producers, the analysis was supported by a tailored genome-scale model and was validated with enzymatic assays performed at different days of the process. A total of 59 reactions were examined from different key pathways, namely glycolysis, pentose phosphate pathway, TCA cycle, lipid metabolism, and oxidative phosphorylation. The intracellular fluxes did not show a metabolic correlation between high producers, but the degree of similitude observed between cell lines could be confirmed with additional experimental observations. The whole analysis led to a better understanding of the metabolic requirements for all the cell lines, allowed to the identification of metabolic bottlenecks and suggested targets for further cell line engineering. This study is a successful application of a curated genome-scale model to multiple industrial cell lines, which makes the metabolic model suitable for process platform.

KW - Genome-scale metabolic model

KW - Chinese hamster ovary

KW - Flux distribution

KW - Mathematical modeling

KW - Metabolic engineering

U2 - 10.1016/j.mec.2019.e00097

DO - 10.1016/j.mec.2019.e00097

M3 - Journal article

VL - 9

JO - Metabolic Engineering Communications

JF - Metabolic Engineering Communications

SN - 2214-0301

M1 - e00097

ER -