A hybrid LBA-LCMS assay for determination of a protein biomarker in monkey serum was developed. The procedure involves magbead-based immunoenrichment followed by tryptic digestion and signature peptide detection by LCMS. The assay was able to demonstrate sub-ng/mL sensitivity. Challenges common to immunoassays were encountered, including minimum required dilution and interference from an anti-analyte antibody present in samples. However, the hybrid nature of the assay allowed novel approaches to overcoming these challenges.
Learning Objectives:
Upon completion, participants will be able to discuss some of the challenges and advantages of analyzing biomarker proteins by LCMS.
Upon completion, participants will be able to understand the principles of using immunoprecipitation and tryptic digestion to a signature peptide for quantitation of a protein in biological samples.
Upon completion, participants will be able to describe the basic aspects of a workflow for developing an LCMS assay for a large molecule protein.
