A Capture Coupling Method for the Covalent Immobilization of Hexahistidine Tagged Proteins for Surface Plasmon Resonance

Surface plasmon resonance (SPR) is a robust method to detect and quantify macromolecular interactions; however, to measure binding interactions, one component must be immobilized on a sensor surface. This is typically achieved using covalent immobilization via free amines or thiols, or noncovalent immobilization using high-affinity interactions such as biotin/streptavidin or antibody/antigen. In this chapter we describe a robust method to covalently immobilize His₆ fusion proteins on the sensor surface for SPR analysis.