HTRF is a homogenous assay that combines fluorescence resonance energy transfer technology (FRET) with time-resolved measurement (TR). It is one of the most frequently used methodologies for drug target studies in high-throughput screening (HTS). In FRET assays, a signal is generated through fluorescent resonance energy transfer between a donor and an acceptor fluorophore when in close proximity one to each other.

FRET can be combined with TR measurements if lanthanides are used as donors, since they show long emission lifetimes allowing for a time delay of the signal detection between the excitation of the donor and the fluorescence of the acceptor fluorophore. For its unique features, in TR-FRET experiments, buffer and media interference is dramatically reduced. HTRF assay sensitivity and robustness make it appropriate for miniaturization in 384 and 1536-well plate formats.

Homogeneous Time Resolved Fluorescence

CELT-335 hCB1/CB2 Cannabinoids receptors fluorescent ligand is suitable to measure the affinity of compounds for the CB1 receptor in competition experiments using HTRF. Shown is a representative example of the HTRF competition binding between CELT-335 and three natural cannabinoids (THC, THCA and THCV).

HTRF validation performed in the Molecular Neuropharmacology Laboratory of the University of Barcelona.

Figure from Lu Raïch et al. “Similarities and differences upon binding of naturally occurring Δ9-tetrahydrocannabinol-derivatives to cannabinoid CB1 and CB2 receptors” Pharmacol. Res., 2021,174,105970. https://doi.org/10.1016/j.phrs.2021.105970

Celtarys Ligands validated for Homogeneous Time Resolved Fluorescence assays

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