From radioactive ligands to biosensors: binding methods with olfactory proteins

In this paper, we critically review the binding protocols currently reported in the literature to measure the affinity of odorants and pheromones to soluble olfactory proteins, such as odorant-binding proteins (OBPs), chemosensory proteins (CSPs) and Niemann-Pick class C2 (NPC2) proteins. The first part contains a brief introduction on the principles of binding and a comparison of the techniques adopted or proposed so far, discussing advantages and problems of each technique, as well as their suitable application to soluble olfactory proteins. In the second part, we focus on the fluorescent binding assay, currently the most widely used approach. We analyse advantages and drawbacks, trying to identify the causes of anomalous behaviours that have been occasionally observed, and suggest how to interpret the experimental data when such events occur. In the last part, we describe the state of the art of biosensors for odorants, using soluble olfactory proteins immobilised on biochips, and discuss the possibility of using such approach as an alternative way to measure binding events and dissociation constants.