Thermodynamic profiling and fragment screening of GPCRs using grating-coupled interferometry.

Area of Science:

• Biochemistry
• Pharmacology
• Biophysics

Background:

• G protein-coupled receptors (GPCRs) are crucial drug targets but often underexplored due to limited knowledge and technical challenges.
• Characterizing GPCRs is essential for advancing drug discovery efforts.

Purpose of the Study:

• To evaluate grating-coupled interferometry (GCI) as a biosensor platform for GPCR research.
• To demonstrate the utility of GCI for kinetic, thermodynamic, and fragment-based drug discovery on GPCRs.

Main Methods:

• Utilized grating-coupled interferometry (GCI) with the human adenosine A2A receptor (A2A R) as a model.
• Employed waveRAPID injection method for kinetic and affinity determination.
• Combined waveRAPID injections with WAVEdelta system for thermodynamic profiling and performed kinetic fragment screening.

Main Results:

• GCI provided high-quality kinetic data comparable to established Biacore technology.
• Demonstrated fast and reliable determination of binding kinetics and affinities using minimal protein.
• Identified specific A2A R binders through kinetic fragment screening, confirmed by nanoDSF.

Conclusions:

• GCI is a versatile and powerful platform for GPCR research, offering rich kinetic and thermodynamic information.
• This approach facilitates early-stage drug discovery by enabling efficient characterization of GPCR targets and ligands.
• GCI supports fragment-based discovery, accelerating the identification of novel drug candidates for GPCRs.