Application of high-resolution MS in the quantification of a therapeutic monoclonal antibody in human plasma. (July 2014) - Mekhssian, Kevork
Background: Monoclonal antibodies are the fastest growing class of protein therapeutics. Ligand-binding assays have been the technique of choice for the quantification of these large proteins; however, LC–MS and more recently LC–HRMS have been gaining momentum as robust alternatives for the bioanalysis of antibodies in biological matrices.Results: Optimization of sample preparation and LC–HRMS analysis in MRM HR mode has allowed us to develop a highly specific dual-peptide targeted assay for the quantification of Rituximab, in human plasma. The linearity of the assay was established from 1.0 to 200 µg/ml for both light and heavy chain surrogate peptides, with accuracy and precision within 15%.Conclusion: LC–HRMS can be an effective tool for the quantification of monoclonal antibodies in regulated bioanalysis.