The objective of this study is to develop expertise in the analytical analyses required for development of a quantitative kinetic model for the rate of carbonyl formation in polypeptides by metal ion-catalyzed oxidations (MCO). The long-term operation of fermentation and cell culture processes and the commercialization of therapeutic proteins have been impeded by stability problems. Oxygen, metals, and reducing agents found in bioprocessing media combine to cause irreversible damage to proteins. The rate constants and parametric sensitivities of this class of oxidation reactions have not been systematically studied under industrially relevant conditions, and highly reactive polypeptide sequences have not been identified. Thus, no quantitative kinetic models exist for use in process design, control, or optimization, nor is sequence information available for protein engineering strategies. This work intends to lay the foundation for the ultimate goal of identification of effective process and/or protein engineering strategies for minimizing oxidative damage.
