This project's objective was to develop a robust, fieldable sensor for the detection of cannabinoids (natural or synthetic) in real time, so as to provide the U.S. Military and law enforcement agencies with a tool to assist in curbing cannabinoid use.
In one effort, the project developed a cannabinoid (CB) receptor-based assay that uses fluorescence resonance energy transfer (FRET) coupling and dark quenchers. Three dark quenchers synthetic cannabinoid (SC) conjugates were developed and characterized. Two of these conjugates showed weak, micromolar binding of the CB receptor; however, the binding was insufficient for a reliable robust sensing platform. In a second effort, a fluorescence-based CB activation assay was developed. Fluorescent molecules tagged to guanine troposphere were bound to the G-protein after receptor binding with SCs. Two fluorescent reporters - Europium-guanine troposphere (Eu-GTP) and boron-dipyrromethene-guanine triphosphate (BODIPY-GTP) - were tested using a high throughput plate reader approach. The BODIPY-GTP assay was the most promising, as it was able to detect four cannabinoid compounds - namely Ä9-tetrahydrocannabinol, JWH307, Win55,212, and CP55,940 - with a 30-percent increase in fluorescence under laboratory test conditions. In its current form, the BODIPY-GTP assay shows the most promise as a high throughput, laboratory- based sensor capable of testing 92 samples in under 10 minutes. 93 figures, 13 tables, 88 references and appended details on the cannabinoid receptor 2 homology model and CHARMM parameter (ParamChem) for Select Cannabinoids
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