Protein Therapeutic Bioanalysis by Hybrid IA-LC-MS/MS:
Optimization of a Preclinical Antibody Workflow

Bioanalysis of protein therapeutics by LC-MS/MS continues to gain popularity as a complementary approach to ligand-binding assays (LBA). LC-MS/MS offers multiple advantages over LBA including higher specificity, multiplexing capability and reduced development time. Immunoaffinity enrichment can be coupled to LC-MS/MS to produce a hybrid method that increases sensitivity and selectivity.

This webinar will highlight the systematic optimization of a flexible, hybrid IA-LC-MS/MS workflow that uses streptavidin-coupled magnetic beads, ’rapid’ trypsin digestion and surrogate peptide quantification.

Key areas of focus will include:

• The selection of streptavidin-coupled magnetic beads as the platform for the immunoaffinity enrichment system due to their amenability to use in combination with analyte-specific biotinylated capture reagents
• A workflow case study for a humanized antibody therapeutic in preclinical serum using generic conserved surrogate peptides for quantification that, when optimized, ultimately requires only ~3 hours with minimal direct handling
• Additional applications for the optimized workflow, such as highly selective and sensitive analysis of humanized antibody in preclinical samples and protein therapeutics using analyte-specific biotinylated capture reagents