김지현 (성신여자대학교)

고병준 (성신여자대학교)

김지현 (성신여자대학교)

고병준 (성신여자대학교)

 Monoclonal antibodies (mAbs) are widely used biopharmaceuticals, and maintaining their structural integrity is essential to ensure their quality, efficacy, and safety. Among the various factors that affect mAb stability, thermal stress is one of the major causes of structural degradation, leading to protein aggregation and post-translational modifications (PTMs) that alter their physicochemical properties and compromise product stability. In this study, bevacizumab was incubated under physiological (37°C) and accelerated thermal stress (45°C) conditions to investigate temperature-dependent aggregation and post-translational modifications (PTMs), two major indicators of antibody structural stability. Protein aggregation was analyzed using SEC-HPLC, and representative samples were subjected to tryptic digestion followed by peptide mapping analysis. Both protein aggregation and PTM abundance increased in a temperature- and time-dependent manner, with structural changes occurring more rapidly under accelerated thermal stress. Oxidation and deamidation were identified as the predominant PTMs, while additional modifications were observed after prolonged heat exposure. These finding improve our understanding of thermal stress-induced structural changes in bevacizumab and provide a basis for the stability assessment of therapeutic monoclonal antibodies.​