Enzyme capacity-based genome scale modelling of CHO cells.
Metab Eng
; 60: 138-147, 2020 07.
Article
in En
| MEDLINE
| ID: mdl-32330653
ABSTRACT
Chinese hamster ovary (CHO) cells are most prevalently used for producing recombinant therapeutics in biomanufacturing. Recently, more rational and systems approaches have been increasingly exploited to identify key metabolic bottlenecks and engineering targets for cell line engineering and process development based on the CHO genome-scale metabolic model which mechanistically characterizes cell culture behaviours. However, it is still challenging to quantify plausible intracellular fluxes and discern metabolic pathway usages considering various clonal traits and bioprocessing conditions. Thus, we newly incorporated enzyme kinetic information into the updated CHO genome-scale model (iCHO2291) and added enzyme capacity constraints within the flux balance analysis framework (ecFBA) to significantly reduce the flux variability in biologically meaningful manner, as such improving the accuracy of intracellular flux prediction. Interestingly, ecFBA could capture the overflow metabolism under the glucose excess condition where the usage of oxidative phosphorylation is limited by the enzyme capacity. In addition, its applicability was successfully demonstrated via a case study where the clone- and media-specific lactate metabolism was deciphered, suggesting that the lactate-pyruvate cycling could be beneficial for CHO cells to efficiently utilize the mitochondrial redox capacity. In summary, iCHO2296 with ecFBA can be used to confidently elucidate cell cultures and effectively identify key engineering targets, thus guiding bioprocess optimization and cell engineering efforts as a part of digital twin model for advanced biomanufacturing in future.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
CHO Cells
/
Enzymes
Type of study:
Prognostic_studies
Limits:
Animals
Language:
En
Journal:
Metab Eng
Journal subject:
ENGENHARIA BIOMEDICA
/
METABOLISMO
Year:
2020
Type:
Article
Affiliation country:
Singapore