title: In situ monitoring of intracellular glucose and glutamine in CHO cell culture.
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.
authors.affiliation: Division of Molecular Biosciences, Department of Life Sciences, Imperial College London, London, United Kingdom.
authors.forename: Alireza
authors.initials: A
authors.lastname: Behjousiar
authors.forename: Cleo
authors.initials: C
authors.lastname: Kontoravdi
authors.forename: Karen M
authors.initials: KM
authors.lastname: Polizzi
mesh_terms: D000818:Animals; D061249:Batch Cell Culture Techniques; D015374:Biosensing Techniques / Q000379:methods*; D016466:CHO Cells; D002138:Calibration; D006224:Cricetinae; D003412:Cricetulus; D005947:Glucose / Q000378:metabolism*; D005973:Glutamine / Q000378:metabolism*; D042541:Intracellular Space / Q000378:metabolism*; D013050:Spectrometry, Fluorescence
chemical_list: D005973:Glutamine; D005947:Glucose