2024. 08.28 (수) ~ 2024. 08.30 (금)
군산새만금컨벤션센터(GSCO)
제목 | High-Resolution Cyclic Ion Mobility-Mass Spectrometry for Analytical Characterization of Therapeutic Antibody |
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작성자 | Abhik Mojumdar (Korea Basic Science Institute) |
발표구분 | 포스터발표 |
발표분야 | 4. Medical / Pharmaceutical Science |
발표자 |
Abhik Mojumdar (Korea Basic Science Institute (KBSI)) |
주저자 | Abhik Mojumdar (Korea Basic Science Institute (KBSI)) |
교신저자 |
Kun Cho (Korea Basic Science Institute (KBSI)) |
저자 |
Abhik Mojumdar (Korea Basic Science Institute (KBSI)) Hee-Jin Yoo (Korea Basic Science Institute (KBSI)) Duck-Hyun Kim (Korea Basic Science Institute (KBSI)) Sun-Hee Choi (Korea Basic Science Institute (KBSI)) Kun Cho (Korea Basic Science Institute (KBSI)) |
Therapeutic proteins exhibit structural diversity due to various modifications during production. Characterization of monoclonal antibodies (mAbs) is crucial for ensuring their quality, efficacy, and safety as therapeutic agents. Combining ion mobility (IM) with high-resolution mass spectrometry (HRMS) offers enhanced structural information, including native gas-phase collision cross-section (CCS), collision-induced unfolding (CIU), and protein-ligand binding, which are typically not provided by HRMS alone. The novel Waters™ SELECT SERIES Cyclic IMS MS platform addresses these complex biophysical challenges. This study comprehensively analyzes adalimumab, trastuzumab, and pembrolizumab, widely used therapeutic mAbs, under both native and denaturing conditions to determine their intact masses. We compared size exclusion chromatography (SEC) and reverse phase liquid chromatography (RPLC) coupled with cIM-MS, highlighting the advantages and limitations of each method in mAb characterization. Structural analyses encompassed primary structure analysis and higher-order structure analysis using advanced mass spectrometry techniques, providing insights into the sequence integrity and conformational stability of the mAbs. Additionally, subunit analysis of the mAbs was performed, providing detailed information on the individual components and their modifications, which is essential for understanding the overall structure and function of the antibodies. Our findings demonstrate that cIM-MS, in conjunction with UPLC, offers superior resolution and sensitivity in analyzing intact mAbs and their structural variants. This comprehensive analytical approach ensures robust evaluation of mAb integrity and stability, supporting the development and quality control of monoclonal antibody therapeutics. |