Development of An Automated Immunopurification Method for The Quantitation of A Therapeutic mAb in Rat Plasma by LC-MS/MS

Bottom-up protein quantitation by surrogate peptide approach with liquid chromatography-tandem mass spectrometry (LC-MS/MS) becomes an increasingly popular field for pharmacokinetics study in drug discovery. Immunopurification (IP) is often utilized to achieve better sensitivity and specificity for LC-MS based protein analysis. However, the complicated sample preparetion procedures of IP and enzymatic digestion lead to high requirements for analyst experience and tedious manual work. Automation would increase method reproducibility and robustness, and reduce manual work. 

Here, we present an automated method for IP and tryptic digestion for protein quantitation with trastuzumab as a model system. In this study, a robust method with IMCStips for IP and ProteinWorks kits for digestion was developed, qualified and applied in a pharmacokinetics study.