The long-term goal of this project is to define a standard methodology for the general high-throughput detection and validation of protein-protein interactions, and provide simultaneous quantitative measurements of binding affinity. 

The first aim of this project is to characterize and define the optimal conditions for Luminescent Resonance Energy Transfer (LRET) arising from a protein engineered in the applicant’s laboratory called TerboFluor.  Preliminary work has shown that this protein forms a novel proximity assay when paired with enhanced Green Fluorescent Proteins (eGFP) and theory suggests that it will be even better with monomeric Red Fluorescent Protein.  Hence this assay may be used detect molecular interaction events.  As TerboFluor may also provide accurate quantification of gene products via titration with Terbium (III), it may be possible to measure binding affinity by accurately determining the quantity of bound and free protein in the same protein interaction assay system.

This work is funded by a grant to CWV Hogue via the NUS Academic Research Fund.


The Terbofluor Patent Application