Differentiating the Effects of Oxidative Stress Tests on Biopharmaceuticals

• DOI: 10.1007/s11095-019-2627-2
• Publication Date: 2019-05-17T19:42:52Z
• Authors: Geoffrey A. Heinzl; Lo Lai; V. Ashutosh Rao
• Keywords: Biological Products; Trastuzumab; Antibodies, Monoclonal, Humanized; Oxidants; Oxidative Stress; 03 medical and health sciences; Antineoplastic Agents, Immunological; Methionine; 0302 clinical medicine; Drug Stability; Cell Line, Tumor; Humans; Indicators and Reagents; Reactive Oxygen Species; Rituximab; Oxidation-Reduction

A rapid and broadly applicable method to assess relevant oxidative damage in biopharmaceuticals is important for lifecycle management of product quality. Multiple methods are currently employed as stress tests to induce oxidative damage for assessment of stability, safety, and efficacy. We compared two common methods for inducing oxidative damage to assess differences in impact on bioactivity and structure of the biopharmaceuticals.Biopharmaceuticals were treated with either metal-catalyzed oxidation (MCO) conditions or the reactive-oxygen species (ROS) inducer 2,2'-Azobis(2-amidinopropane) dihydrochloride (AAPH), then analyzed for changes in structure and bioactivity.We demonstrate that commonly used chemical methods for assessing oxidation yield distinct oxidation profiles for each of the biotechnology products analyzed, including monoclonal antibodies. We further report oxidant- and product-specific changes in bioactivity under oxidizing conditions, along with differential oxidation on the molecular subunits of monoclonal antibodies.Our results highlight the need for product-specific optimization and selection of orthogonal, relevant oxidizers when characterizing stress responses in biopharmaceuticals.