Hybrid LC-MS assays, combining features from both ligand binding for preferred analyte enrichment and LC-MS for enhanced detection selectivity, have demonstrated the potential as a complementary or alternative platform to conventional LBAs for bioanalysis of protein therapeutics and biomarkers. This novel technology effectively allows characterization and quantification of biological target analytes, including mAbs, ADCs, other complex modalities and tissue biomarkers where no suitable solutions existed previously.

This specific workshop is aimed to provide the audience a general introduction, as well as recent developments, of this powerful analytical technology. Both bottom-up and top-down approaches in gaining quantitative and qualitative (e.g., major PTMs) information of protein analytes will be described. Advantages and certain limitations associated with this new platform will be discussed. Recent progress in this field will be highlighted to demonstrate how a variety of analytical challenges are overcome to pave the way for an increasing practical applications of hybrid LC–MS in characterization and quantification of protein therapeutics and biomarkers. Case studies will include immunoaffinity enrichment, protein biotransformations by HRMS, quantification of total antibody and antibody-conjugated payload of ADCs, tissue-based bioanalysis, LC/MS in CMC for specification tests of APIs, considerations for improving the assay sensitivity, as well as outsourcing.