Matsuda Yutaka, Kliman Michal, Mendelsohn Brian A
Ajinomoto Bio-Pharma Services, 11040 Roselle Street, San Diego, California 92121, United States.
Waters Corporation, 34 Maple Street, Milford, Massachusetts 01757-3696, United States.
J Am Soc Mass Spectrom. 2020 Jul 1. doi: 10.1021/jasms.0c00129.
Antibody-drug conjugates (ADCs) are at the forefront of the next generation of oncology biopharmaceuticals. Conventional ADCs involve stochastic conjugation of the antibody to a cytotoxic drug, creating a highly heterogeneous product. The resulting stochastic distribution often leads to a narrow therapeutic index and makes it difficult to analyze the composition of heterogeneous ADCs. With the goal of overcoming these issues, we developed a site-specific conjugation technology, named AJICAP, for production of low heterogeneity ADCs. For analysis of these site-specific ADCs, we report herein strong cation exchange chromatography coupled with UV and mass spectrometry detection (SCX-UV-MS). Retention time reproducibility after SCX column equilibration enabled monitoring of important changes in product quality. SCX-UV-MS performed with MS-compatible mobile phases was conducted for intact native ADC analysis, allowing drug-antibody ratio characterization and charge variant characterization in single analysis. Furthermore, subunit analysis of the site-specific ADCs by native SCX-UV-MS confirmed the Fc site selectivity of ADCs generated by AJICAP conjugation.
抗体药物偶联物（ADCs）处于下一代肿瘤生物制药的前沿。传统的ADCs涉及抗体与细胞毒性药物的随机偶联，产生高度异质性的产物。由此产生的随机分布通常导致治疗指数狭窄，并且难以分析异质性ADCs的组成。为了克服这些问题，我们开发了一种名为AJICAP的位点特异性偶联技术，用于生产低异质性的ADCs。为了分析这些位点特异性ADCs，我们在此报告了结合紫外和质谱检测的强阳离子交换色谱法（SCX-UV-MS）。SCX柱平衡后的保留时间重现性能够监测产品质量的重要变化。使用与质谱兼容的流动相进行的SCX-UV-MS用于完整天然ADCs的分析，允许在单次分析中表征药物-抗体比率和电荷变体。此外，通过天然SCX-UV-MS对位点特异性ADCs进行亚基分析，证实了由AJICAP偶联产生的ADCs的Fc位点选择性。
