Bio-nanotechnology and nanoscale analysis are becoming increasingly prevalent, raising the demand for techniques with high sensitivity that can detect biomolecules in biological samples. Bioluminescent photoproteins, such as aequorin, possess great potential to provide with a solution to this new challenge because they can be detected at low concentrations, and have different emission wavelengths depending on the protein variant used, a property that can be exploited in multiplex analysis. In our laboratory we design and prepare new aequorin variants to broaden the scope of their use in bioanalysis, thus allowing for detection of biomolecules that are not measurable by other technologies. Protein molecular switches with optical properties are another type of designer biomolecules that, in the presence of a target ligand, demonstrate an altered response manifested by an “on/off” signal.
Specifically, we design and develop bioluminescent molecular switches that incorporate the recognition properties of binding proteins with the bioluminescence afforded by the aequorin variants. In addition, we are investigating the use of a series of computational and synthetic approaches along with genetic engineering strategies to prepare new bioluminescent proteins and molecular switches with a wide range of spectral properties. These molecules can provide with new enabling technologies forin vitroandin vivobiosensing, imaging, and multiplex analysis that have a number of advantages over existing methods.