mTR-FRET™ Technology

Single-label technology

Our technology for protein detection and protein-ligand interactions is based on the mTR-FRET™ technology, making use of single-label approach in a homogeneous high throughput compatible assay format. The novel proprietary single-label method with time-resolved fluorescence (TRF) readout requires a ligand, such as small molecule, peptide or aptamer, conjugated with our lanthanide chelates, while the target protein is intact, unlabeled. A proprietary assay buffer modulates TRF-signal of non-bound lanthanide-ligand enabling direct measurement of bound labelled ligand-receptor or ligand-cell complex.

By providing the technology to assay proteins without labelling with the single-label ligand-labeled strategy is highly cost-effective in a mix-and-measure assay format. This enables researchers to investigate a wider range of target proteins compared to dual-label or surface-immobilization strategies. The method eliminates the labelling of the target protein, providing a user-friendly format to develop assays in a microliter assay volumes.

The single-label mTR-FRET™ assay technology can be developed for small-molecule, peptide and aptamer binding to target protein. Successful mTR-FRET™ assays are based on our lanthanide chelates and propriety assay modulation solutions.
TR-FRET Technology

Dual-label technology

Förster resonance energy transfer (FRET) describes a physical phenomenon of non-radiative energy transfer based on dipole-dipole coupling that can occur from an excited state fluorophore (the donor) to a ground state fluorophore (the acceptor). TR-FRET (time-resolved FRET) is a homogeneous technique, which combines standard FRET technology with time-resolved fluorescence (TRF) measurement, eliminating short-lived background fluorescence. TR-FRET is a well-established technology and it is currently widely used in drug discovery, clinical diagnostics, and academia. As a highly versatile assay technology, TR-FRET allows the study of a wide range of low or high affinity biological interactions, using either small or large molecules.

TR-FRET is typically performed as a sandwich-type assay with two labeled binders, or labeled analyte and binder, for example antibody. TR-FRET exploits the unique photophysical properties of lanthanide labels when used as TR-FRET donor, and far-red acceptors. These properties enable essentially zero background signal and high S/B ratio, two features critical for the development of reproducible and robust assays. TR-FRET assays are simple to perform by combining the analyte/sample and detection reagents without any separation step. After incubation, result can be read by using TR-FRET compatible or certified microplate reader.

The dual-label TR-FRET assay technology can be developed for a wide range of molecules including proteins and thus it supports our mTR-FRET™ technology. Successful TR-FRET assays are based on carefully selected donor-acceptor pairs showing exceptional spectral compatibility and TR-FRET signal.
mTR-FRET – single-label detection:
Lanthanide-chelate labelled small molecule (Ln) binds to target protein. Target-protein bound labeled-molecule yields high TRF signal while non-bound Ln-labelled small molecule is quenched in the detection solution 
TR-FRET – dual-label detection:
Acceptor-dye labelled small molecule/protein binds to target-protein labelled with lanthanide-chelates. Ln-chelates of the target-protein transfer energy to the target-bound acceptor-labelled small molecule/protein providing high TR-FRET signal. Non-bound acceptor-labelled small molecule/protein lacks the proximity and no energy is transferred from Ln-labelled target protein.