Abstract
Background A thorough knowledge of the metabolism and its compartmentation in mammalian cells is desirable to enable rational design and optimization of producing cell lines and production processes for biopharmaceuticals. In this study we focused on acquiring a detailed understanding of metabolite channelling and the metabolic flux distribution during overflow metabolism in the human cell line AGE1.HN (ProBioGen AG). This metabolic phenotype characterized by energy spilling as well as waste product formation is commonly observed in the beginning of the cultivation [1]. C tracer experiments and C flux analysis as applied in this investigation represent methods offering in-depth insights into cellular physiology [2,3].
Highlights
A thorough knowledge of the metabolism and its compartmentation in mammalian cells is desirable to enable rational design and optimization of producing cell lines and production processes for biopharmaceuticals
Glucose carbons were entering the tricarboxylic acid (TCA) cycle which can be deduced from an increase in fractional labelling of glutamate and proline in glucose tracer experiments
Reflux from TCA cycle metabolites to glycolytic metabolites was detected since labelling in lactate as well as alanine was measured when using [U-13C5] glutamine as tracer
Summary
A thorough knowledge of the metabolism and its compartmentation in mammalian cells is desirable to enable rational design and optimization of producing cell lines and production processes for biopharmaceuticals. In this study we focused on acquiring a detailed understanding of metabolite channelling and the metabolic flux distribution during overflow metabolism in the human cell line AGE1.HN® (ProBioGen AG). This metabolic phenotype characterized by energy spilling as well as waste product formation is commonly observed in the beginning of the cultivation [1]. 13C tracer experiments and 13C flux analysis as applied in this investigation represent methods offering in-depth insights into cellular physiology [2,3] This metabolic phenotype characterized by energy spilling as well as waste product formation is commonly observed in the beginning of the cultivation [1]. 13C tracer experiments and 13C flux analysis as applied in this investigation represent methods offering in-depth insights into cellular physiology [2,3]
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