Effect of amino acid supplementation on titer and glycosylation distribution in hybridoma cell cultures-Systems biology-based interpretation using genome-scale metabolic flux balance model and multivariate data analysis

• Published in: Biotechnology progress Vol. 32; no. 5; pp. 1163 - 1173
• Main Authors: Reimonn, Thomas M., Park, Seo-Young, Agarabi, Cyrus D., Brorson, Kurt A., Yoon, Seongkyu
• Format: Journal Article
• Language: English
• Published: United States Blackwell Publishing Ltd 01.09.2016; Wiley Subscription Services, Inc
• Subjects: Amino acids; Amino Acids - chemistry; Amino Acids - metabolism; Animals; cell culture; Cell Culture Techniques; Cells, Cultured; dynamic flux balance analysis; Fluctuations; Glycosylation; hybridoma; metabolomics; Mice; Models, Biological; Multivariate Analysis; Objective function; Systems Biology; systems biology; dynamic flux balance analysis; hybridoma; cell culture; metabolomics
• ISSN: 8756-7938; 1520-6033; 1520-6033
• DOI: 10.1002/btpr.2335

Genome‐scale flux balance analysis (FBA) is a powerful systems biology tool to characterize intracellular reaction fluxes during cell cultures. FBA estimates intracellular reaction rates by optimizing an objective function, subject to the constraints of a metabolic model and media uptake/excretion rates. A dynamic extension to FBA, dynamic flux balance analysis (DFBA), can calculate intracellular reaction fluxes as they change during cell cultures. In a previous study by Read et al. (2013), a series of informed amino acid supplementation experiments were performed on twelve parallel murine hybridoma cell cultures, and this data was leveraged for further analysis (Read et al., Biotechnol Prog. 2013;29:745–753). In order to understand the effects of media changes on the model murine hybridoma cell line, a systems biology approach is applied in the current study. Dynamic flux balance analysis was performed using a genome‐scale mouse metabolic model, and multivariate data analysis was used for interpretation. The calculated reaction fluxes were examined using partial least squares and partial least squares discriminant analysis. The results indicate media supplementation increases product yield because it raises nutrient levels extending the growth phase, and the increased cell density allows for greater culture performance. At the same time, the directed supplementation does not change the overall metabolism of the cells. This supports the conclusion that product quality, as measured by glycoform assays, remains unchanged because the metabolism remains in a similar state. Additionally, the DFBA shows that metabolic state varies more at the beginning of the culture but less by the middle of the growth phase, possibly due to stress on the cells during inoculation. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1163–1173, 2016