Measure the functional potency (EC50) and efficacy (Emax) of agonists at a Gs-coupled GPCR by quantifying intracellular cAMP accumulation with a homogeneous time-resolved FRET (HTRF) readout in a stably transfected cell line. The goal is to generate concentration-response curves for agonist pharmacology and partial-agonist classification.
Confirm direct, intracellular target engagement of a small-molecule inhibitor by measuring ligand-induced thermal stabilization of the endogenous target protein in intact A549 cells, generating melt curves and an isothermal dose-response (ITDRF) to estimate cellular engagement potency.
Flag cardiac ion-channel and general cytotoxicity liabilities of lead compounds by combining a thallium-flux hERG potassium-channel inhibition assay in hERG-stable HEK293 cells with a parallel ATP-based viability counter-screen, generating IC50 (hERG) and CC50 (cytotoxicity) values to compute a safety margin.
Determine the half-maximal inhibitory concentration (IC50) of small-molecule inhibitors against a recombinant serine/threonine kinase using a luminescent ADP-detection format. The goal is to rank-order compounds by potency and generate reproducible, four-parameter logistic curves suitable for SAR decisions.
Determine the in vitro metabolic stability of lead compounds in pooled human liver microsomes by measuring NADPH-dependent substrate depletion over time, and calculate intrinsic clearance (CLint) and predicted hepatic clearance to triage compounds by metabolic liability.