Functional assay for screening GPCR targets

BRET-based biosensors for predicting therapeutic efficacy and side effects of your leads

BRET (bioluminescence resonance energy transfer) based technology is the system of choice for monitoring in real time intermolecular/intramolecular events (protein-protein interactions, conformational changes within a protein or protein-complexes) in living cells.

BRET is suitable for:

  • Hit to lead and lead optimization: screening/identification/characterization of pharmacological ligands efficacy (agonists, antagonists, inverse agonists, partial agonists…)
  • Receptor research (activation, inactivation, dimerization, internalization..): Tyrosin kinase and G protein-coupled receptors (GPCRs)
  • Mapping of signalling pathways: protein translocation (Calcium / Calmoduline complexe formation), ubiquitination, autophagic process, kinase activity (cAMP binding and subsequent activation of protein kinase A), second messenger production (cAMP and Ca2+ pathways)

Advantages

  • Direct, real time and rapid investigation
  • Manufactured biosensors
  • Highly sensitive and quantitative assay
  • Applicable to established stable cell lines or primary culture

Available services

Based on an unparalleled expertise in BRET based technology, we can perform:

  • Screening with a panel of biosensors (GPCRs, beta arrestin, Calcium/Calmodulin-dependent kinase…)
  • Manufacture of custom designed biosensors to target your needs
  • Target validation
  • Lead optimization
  • Mechanistic studies
  • R&D program to develop biosensors monitoring specific signaling pathways

Contact us for more information on this service.

Functional assay for screening GPCRs

BRET principle

BRET assay technology is based on a naturally occurring nonradiative resonance energy transfer event between an enzymatic energy donor (RLuciferase) and a fluorescent energy acceptor (GFP-derived fluorophores).

Appropriate energy donor and acceptor molecules are either fused to target proteins that might interact with each other (intermolecular BRET) or fused within a unique protein subject to conformational changes (intramolecular BRET).

The proximity of these candidates can then be measured by fluorescence resonance energy transfer from the donor to the acceptor. Thus, the presence or absence of the BRET signal acts as a “molecular yardstick” (see illustration below).

Bret signal
 
ipsum ut diam sed pulvinar facilisis elementum elit. commodo eleifend at