(T0930-11-62) Strategic Optimization and Evaluation of Stress Conditions for IgG4 Antibodies

Purpose: Establish a benchmark stress study framework by optimizing stress conditions using IgG4 antibodies.

Methods: An IgG4 antibody was exposed to a range of oxidative, low pH, high pH, and photo stress conditions. Stressed samples were analyzed by a series analytical assays such as SEC, cIEF, CE-SDS, LCMS, and SoloVPE. These assays provided a comprehensive dataset on the physical and chemical characteristic of the antibody under each stress condition, facilitating the selection of optimized conditions for each stress arm.

Results: An IgG4 antibody was used as a model to determine the primary degradation pathways which exposed to pH, oxidation, and photo stress conditions. In the oxidation arm, exposure to varying concentrations of tert-butyl hydroperoxide led to the notable oxidation of specific methionine residues without inducing aggregation. Low pH conditions induced increased high molecular weight species (HMWS) with time of exposure. High pH conditions highlighted deamidation as the main chemical degradation pathway. Photodegradation studies designed to mimic manufacturing light conditions, pinpointed oxidation as the primary degradation pathway without substantial impact on physical stability.

Conclusion: This comprehensive study successfully determined optimized stress conditions for each stress arm, unveiling specific degradation pathways for the model IgG4 antibody. These findings offer valuable insights for future IgG4 antibody development including assignment of CQAs, building structure/function knowledge as well as contributing to analytical development.

Acknowledgements: Scott Dooley, Karen Lee, Marc Verhagen