Application of high-resolution MS in the quantification of a therapeutic monoclonal antibody in human plasma

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 MRMHR 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.

n the last few years, therapeutic monoclonal antibodies (mAbs) have emerged as one of the most important drug classes in the biopharmaceutical industry. At present, more than 30 mAbs are marketed for a wide range of therapeutic areas such as oncology, autoimmunity, infectious diseases and metabolic disorders [1,2]. The increasing importance of protein therapeutics necessitates performance improvements in the bioanalytical methods used in the pharmacokinetic assessments at the preclinical and clinical stages of drug development.

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