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Förslaget inkom 2009-06-01

Development of cultivation medium nutrient cocktail by metabolic reaction network – Part 3

OBS! ANSÖKNINGSTIDEN FÖR DETTA EXJOBB HAR LÖPT UT.
The cultivation of Chinese Hamster Ovary cells, CHO, in cultivation medium in absence of animal-derived components is widely used in the biopharmaceutical industry for the production of drug proteins. Cultivation media are complex cocktails with up to 70 components including nutrients, growth factors, vitamins, salts, trace elements, etc. The development of these media is mostly based on empirical or statistical approach. The nutrients, about 20 to 30 components, include amino acids and biosynthesis precursors or intermediates.
The purpose of the present work is to develop a cultivation medium nutrient cocktail by using a model of the metabolic pathways. A CHO cell line representative of industrial cell lines used for the production of biopharmaceuticals, i.e. DHFR- system, will be used. A model of the metabolic pathways is used exploiting the information from the metabolic flux analysis. Cultivation experiments are performed in order to identify the values of the model parameters for the cell line used. Finally the identified model is experimentally verified and exploited to determine an optimal nutrient cocktail. This nutrient optimization is applied to obtain a base medium and a feed strategy minimizing the cellular production of toxic by-product like lactate and ammonium. The cultivation experiments are performed in screening scale, i.e. 1 to 5 mL cultures, shake flasks. Amino acid analysis is performed to support the cultivation.


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