Abstract: New Perspectives for Protein Immunotherapy and Vaccine Development through Antibody Epitope Identification using Affinity- Mass spectrometry
Analysis of biomolecular interaction epitopes has recently become a key stepAnalysis of biomolecular interaction epitopes has recently become a key stepin the development and molecular evaluation of therapeutic antibodies, biomedicalpeptide and protein biomarkers and molecular vaccines. Bioaffinity analysis usingbiosensors has become an established technique for detection and quantification ofbiomolecular interactions. However, a principal limitation of biosensors is their lack ofproviding chemical structure information of affinity-bound ligands. Proteolyticexcision/extraction (PROTEX-MS), hydrogen-deuterium exchange (HDX-MS) of peptidebackbone hydrogens, and Fast- Photochemical Oxidation (FPOP) are major techniques formass spectrometry based elucidation of protein- ligand interactions, but none of these toolsalone provide quantitative affinity data. Using a surface plasmon resonance (SPR) biosensor,we have developed a continuous online biosensor-MS combination with electrosprayionization mass spectrometry that enables the simultaneous affinity isolation, structureidentification and affinity quantification of biopolymer ligands from a protein- ligand compleximmobilized on a gold chip.