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

Speaker

Brendan Powers, PhD, Staff Scientist, Covance

Dr. Brendan Powers joined Covance Bioanalytical in 2018 and has more than ten years in biological and analytical research. In his current role as Staff Scientist, Dr. Powers undertakes original research that includes developing and confirming highly sensitive, reliable assay methodologies for fast and accurate analysis of pharmaceuticals and biopharmaceuticals in biological fluids and tissues. In addition, he is a subject matter expert and global innovation leader for LC-MS analysis of biopharmaceuticals. His key areas of expertise include hybrid IA-LC-MS/MS, regulated LC-MS/MS, molecular biology and large molecule bioanalysis.

Dr. Powers is an author or co-author of more than a dozen peer-reviewed publications and industry presentations. His work can be seen in several leading publications, including the Journal of Cell Science and the Journal of Biochemistry and Molecular Cell, among others. He holds a BS in Biochemistry from the University of Illinois and a PhD in Biochemistry from Purdue University.

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