Antibody oxidation and impact of formulation: A high-throughput screening approach

Oxidation is a complex degradation pathway in biopharmaceutical products that necessitates comprehensive assessment to ensure product stability and safety. This study focuses on implementing an oxidative profiling workflow within a high-throughput (HT) formulation screening process to identify and mitigate potential oxidation liabilities. To assess the feasibility of integrating oxidative stress testing into HT formulation development, we analyzed the oxidation susceptibility of three monoclonal antibodies by varying several formulation parameters including protein concentration, buffer system and pH, surfactant type and concentration as well as presence of antioxidative excipients. Oxidative stress was induced using visible light, hydrogen peroxide, and metal-catalyzed oxidation. HT analytical methods such as Size Exclusion Chromatography and Reversed-Phase Chromatography subunit analysis were employed to assess aggregation and modification of Fc and Fab subunits. An oxidation scoring tool was developed to simplify the evaluation of large datasets. The results demonstrated that formulation composition can significantly influence oxidation susceptibility. However, the outcomes varied greatly among the different antibodies, highlighting the need for a comprehensive profiling approach. The study confirms that the oxidation profiling workflow is an effective method for routine HT formulation screenings, providing a thorough evaluation of the oxidative stability of biopharmaceutical formulations.