Elucidation of the mechanism of physicochemical stress-induced aggregation of therapeutic antibodies and development of a theory to predict aggregation

2021 Fiscal Year Final Research Report

• Project/Area Number: 19H03363
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Review Section: Basic Section 47020:Pharmaceutical analytical chemistry and physicochemistry-related
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: Honda Shinya 国立研究開発法人産業技術総合研究所, 生命工学領域, 副研究部門長 (50344122)
• Project Period (FY): 2019-04-01 – 2022-03-31
• Keywords: タンパク質 / 凝集 / バイオ医薬品 / モノクローナル抗体

Outline of Final Research Achievements

The purpose of this study was to elucidate the mechanism of physicochemical stress-induced aggregation of therapeutic antibodies and to develop a theory to predict aggregation. As a result, we succeeded in identifying the site of local structural change of degraded antibodies induced by acid stress, and revealed that the exposure of the hydrophobic core associated with a slight conformational change in the C-terminus is a common sign of structurally degraded antibodies. The spatial displacement of the domains of degraded antibodies was also quantified, and it was found that at pH 2, the homodimer of the CH3 domain dissociates into single monomers and the native-like quaternary structure is completely disrupted. Furthermore, we performed non-dehydrated and unstained measurements of micrometer-scale aggregates composed of degraded antibodies and confirmed they possess the same fractal dimension as nanometer-scale aggregates.

Free Research Field

タンパク質化学

Academic Significance and Societal Importance of the Research Achievements

本研究で蓄積された治療用抗体の凝集化メカニズムに関する知見は、凝集化予測技術や防止技術の基盤となる。バイオ医薬品の製造管理におけるknowledge-basedからscience-basedへの展開を加速し、治療用抗体の有効性と安全性に関するリスク管理の高度化に寄与することが期待される。また、得られた知見以外にも、本研究で採用した多角的アプローチは、動的な生体高分子の構造状態解析、液中試料の無乾燥・無染色高解像度計測、マルチスケールコロイド科学現象の解明など、関連分野への学術的波及が期待される。