Intact LC-HRMS for bioanalysis is an emerging area of interest and offers potential advantages compared to surrogate peptide approaches. Reduced method development time can be achieved since there is no need to identify or optimize surrogate peptides. Standardized LC and HRMS methods can be applied based on the class of molecule being examined. Intact HRMS also provides an information rich dataset that can provide qualitative information on biotransformation and catabolism. Furthermore, while not as sensitive as surrogate peptide approaches, multiple examples have shown assay LLOQ’s of approximately 50 – 100’s of ng/mL, which are adequate for pharmacokinetic assessment of typical mAbs. Data generated within Amgen using an Agilent AssayMAP Bravo system for immunoaffinity extraction and a Q-Exactive LC-HRMS platform will be presented. LLOQ’s of 50 – 150 ng/mL were obtained from 30 µL of mouse plasma with potential application to cassette dosing.
Learn how intact protein quantification by hybrid LC-HRMS can simplify method development workflows compared to surrogate peptide based approaches.
Demonstrate method performance with and without the use of a surrogate intact protein internal standard.
Demonstrate strategy for obtaining adequate sensitivity (~50 – 150 ng/mL LLOQ) and explore potential application to cassette dosing.