New manuscript accepted
Quantitative characterization of metabolism and metabolic shifts during growth of the new human cell line AGE1.HN using time resolved metabolic flux analysis
Niklas J, Schräder E, Sandig V, Noll T, Heinzle E (2010),
Bioprocess and Biosystems engineering, accepted
For the improved production of vaccines and therapeutic proteins a detailed understanding of the metabolic dynamics during batch or fed-batch production is mandatory. A dynamic metabolic flux analysis method that is well suited to characterize such dynamics is described and was applied to the new human cell line AGE1.HN. Growth and metabolism of AGE1.HN during batch cultivation was changing over time showing three distinct phases characterized by specific differences in the metabolism. Main events that forced an adaptation of cellular metabolism were depletion of pyruvate and glutamine which led to interesting metabolic shifts. These are described in detail showing the reaction of this human cell line to changes in the environment. Important targets of improvement identified from the analysis are (i) the reduction of the overflow metabolism in the beginning of the cultivation, (ii) reduction of pyruvate content in the medium and (iii) prolongation of phase 2 in which the cellular energy metabolism was highly efficient through specific feeding strategies. Generally, this cell line grows to high cell density already in batch culture and produces little overflow metabolites. The method presented allows fast and reliable MFA during the development of producer cells and production processes from microtiter plate to large scale reactors with moderate analytical and computational effort. It is well suited to guide media optimization and genetic engineering of producing cell lines.