Analysis of biomolecular interactions by surface plasmon resonance

Ladislav Bumba

Institute of Microbiology, Czech Academy of Sciences, Videnska 1083, 142 20 Prague 4, Czech Republic

Surface Plasmon Resonance (SPR) is a physical process that can occur when plane-polarized light hits a metal film under total internal reflection conditions. This phenomenon is employed as an excellent technique to measure biomolecular interactions in real-time in a label free environment. In this method, one of the interacting partners (ligand) is immobilized to the sensor surface, while the other (analyte) is free in solution and passed over the surface. The kinetic parameters of the interactions are further calculated from association and dissociation curves. This lecture will introduce a SPR optical biosensor “ProteOn XPR36 protein interaction array system“ recently installed at the Institute of Microbiology in Prague. This system generates a 6 x 6 interaction array for the simultaneous analysis of up to six ligands with up to six analytes which enables analysis of up to 36 different protein interactions in a single run on a single chip. Few examples, such as protein-protein interaction of adenylate cyclase toxin (CyaA) with integrin receptor CD11b/CD18, lipid-protein interaction of phosphatidylinositol 4,5-bisphosphate (PIP2) with membrane channel TrpV1, or DNA-protein interaction of transcription factor FoxO4 with target DNA, will be presented to show the throughput, flexibility and versatility of this instrument.