Ammonium ion metabolism in cultured mammalian cells. Effects on energy metabolism and expression of recombinant complex glycoproteins

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Project objective

The objective of this work is to develop strategies for controlling, in continuous mammalian cell lines, the expression of human-type glycoproteins used as biopharmaceuticals. Investigation will focus on parameters affecting the final oligosaccharide structure and hence the biological properties of recombinant human glycoproteins. The study is based on the novel hypothesis that the intracellular pool of UDP-N-acetylhexosamines (UDP-HexNAc) affects the expression of Nglycans. This pool is modulated by the culture environment, as both glutamine (via ammonium ion release) and glucose can contribute to it. Specific aims of the proposed project include: (a) Characterization of the effects of chemical parameters affecting UDP-HexNAc pool on the carbohydrate expression of a model glycoprotein produced by recombinant mammalian cells. (b) Evaluation of the effect of a shift in intracellular pH or culture temperature on carbohydrate expression. (c) Elucidation of the mechanisms responsible for changes in carbohydrate expression patterns resulting from changes in physical and chemical parameters.

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Participants

• Aristotle University of Thessaloniki · ThessalonikiCoordinatorGreece

Links

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