MEANS AND METHODS FOR BIOLUMINESCENCE RESONANCE ENERGY TRANSFER (BRET) ANALYSIS IN A BIOLOGICAL SAMPLE

摘要

The invention relates to the field of in vitro detection methods using luminescence. Provided is a sensor molecule for detecting an analyte of interest in a sample using bioluminescence resonance energy transfer (BRET), the sensor molecule comprising a proteinaceous moiety tethered to a synthetic regulatory molecule. Also provided is an analytical device comprising a sensor and methods using the sensor molecule.

主权项

1. A sensor molecule for detecting an analyte of interest in a sample using bioluminescence resonance energy transfer (BRET), the BRET sensor molecule comprising a proteinaceous moiety tethered to a synthetic regulatory molecule, wherein
(i) the proteinaceous moiety comprises a luciferase enzyme (Luc) attached to binding protein (BP) capable of binding the analyte of interest; (ii) the synthetic regulatory molecule comprises a ligand (L) capable of intramolecular binding to BP, and a fluorescent acceptor that can accept the energy from the Luc through resonance energy transfer in the presence of the appropriate Luc substrate, and (iii) wherein the binding of analyte to BP changes the degree of intramolecular binding of L to BP of the BRET sensor molecule, thereby resulting in a change in BRET efficiency.