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PLoS One. 2012;7(4):e34512. doi: 10.1371/journal.pone.0034512. Epub 2012 Apr 3.

In situ monitoring of intracellular glucose and glutamine in CHO cell culture.

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  • 1Division of Molecular Biosciences, Department of Life Sciences, Imperial College London, London, United Kingdom.

Abstract

The development of processes to produce biopharmaceuticals industrially is still largely empirical and relies on optimizing both medium formulation and cell line in a product-specific manner. Current small-scale (well plate-based) process development methods cannot provide sufficient sample volume for analysis, to obtain information on nutrient utilization which can be problematic when processes are scaled to industrial fermenters. We envision a platform where essential metabolites can be monitored non-invasively and in real time in an ultra-low volume assay in order to provide additional information on cellular metabolism in high throughput screens. Towards this end, we have developed a model system of Chinese Hamster Ovary cells stably expressing protein-based biosensors for glucose and glutamine. Herein, we demonstrate that these can accurately reflect changing intracellular metabolite concentrations in vivo during batch and fed-batch culture of CHO cells. The ability to monitor intracellular depletion of essential nutrients in high throughput will allow rapid development of improved bioprocesses.

PMID:
22509313
[PubMed - indexed for MEDLINE]
PMCID:
PMC3317985
Free PMC Article

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