Separation techniques for intact antibody analysis by mass spectrometry

• DOI: 10.1080/10826076.2023.2199328
• Publication Date: 2023-04-17T18:15:26Z
• Authors: Nicole A. Schneck; Kshitij Khatri; Matthew D. Maust; Mark E. Jennings; Julien Peltier; John F. Kellie
• Keywords: Biopharmaceutical; Separation method; Ion-mobility spectrometry

The possibilities to directly couple different separation methods with mass spectrometry (MS) for the analysis of intact proteins has attracted significant attention over last decade. While sample preparation is critical MS analyses, coupling an efficient method before can significantly improve ability resolve protein heterogeneity and reveal masses or higher-order structure information (e.g., PTM conformation changes). To that end, this review focuses on common online techniques coupled detection detect antibodies large a focus biopharmaceutical applications. An overview liquid chromatography modes, such as reversed-phase, size-exclusion, mixed-mode, hydrophobic interaction, ion-exchange will be discussed, along electrophoretic separation, gas-phase using ion-mobility other next-generation tools. Finally, application space within industry discussed how improvements ultimately help characterize charge-, size-, variants novel biotherapeutics throughout all stages drug development.