2.12 Surface Plasmon Resonance in Binding Site, Kinetic, and Concentration Analyses
Surface Plasmon Resonance is a biosensor technique that has been refined to provide an insight into the binding behavior of antibody-antigen pairs, which is at the heart of every immunoassay. In this chapter the relative importance of the association and dissociation binding rates, and how they can be visualized, are explained.
Abstract
Surface plasmon resonance is a label-free biosensor technique that has been refined, particularly by BiaCore, to provide a convenient tool for studying protein binding characteristics. This is an illuminating method for the study of antibody binding properties for immunoassay and biopharmaceutical development. The principles are explained, including immobilization and capture techniques, and the interpretation of sensorgrams to visualize antibody on and off rates. Epitope mapping and kinetic analysis are explained. Concentration analysis using SPR is also explained, including calibration-free concentration analysis.
Contributor
Robert Karlsson is currently a staff scientist in Bio-Analysis Systems at GE Healthcare Life Sciences. Robert graduated from the University of Stockholm in 1976. He started his professional career as an Analytical Chemist in Pharmacia later the same year. In the 1980s he was recruited to a project in Pharmacia to develop an SPR biosensor. This resulted in the launch of the first BiacoreTM system in 1990. He published the original paper on kinetic analysis using Surface Plasmon Resonance in 1991 and has since published close to 40 papers describing Biacore applications. Prior to being a staff scientist Robert held positions as senior scientist, group manager and research director. As a research director Robert was responsible for biochemistry, systems, applications and scientific market support teams. Today Robert is engaged with label-free and biomolecular imaging technologies.
Keywords
Surface plasmon resonance, biosensors, protein binding, binding site, monoclonal antibody, affinity, avidity, kinetics, equilibrium constant, association constant, dissociation constant, on rate, off rate, sensorgram, epitope mapping, therapeutic antibodies.
