Mammalian cells, including CHO cells, require complex media for cell growth and protein production. Amino acids are the major components of culture media, classified as essential amino acids that cannot be synthesized do novo by mammalian cells (e.g. valine, phenylalanine) and non-essential amino acids (e.g. aspartate, alanine). Currently, very little is known about the metabolism/catabolism of amino acids in cell culture. In previous work, our group and others have demonstrated that CHO cells take up significantly larger amounts of amino acids than is strictly required for cell growth and protein production. The objective of this project is to apply advanced 13C-flux analysis tools to elucidate the metabolic flow of all 20 amino acids in CHO cells in fed-batch culture. Results from this work will inform what fraction of each amino acid is used for cell growth, protein production, lactate formation, and energy generation at different stages during a culture.

How the Project May be Transformative and/or Benefit Society

This study will elucidate, for the first time, the fate of all amino acids in CHO cell culture and quantify how changes in amino acid profiles translate to changes in metabolism and physiology of CHO cells. No such information exists currently.

Site: University of Delaware and Johns Hopkins University

Project Leaders: Maciek Antoniewicz, Mike Betenbaugh