Project Examples – Development of a GPCR Cell-based Screening Assay

Objective: To develop nonradiometric, robust, and durable agonist assay for the activation of a target GPCR and a highly related GPCR.

Performance Specification: Dose-response with known agonist tool compound

Proposed Timeline: 3 months

Description: IBA took advantage of the fact that many activated GPCRs are coupled to the activation of phospho-MAPK1/MAPK2.  The two target GPCR were stably expressed in cell culture and assessed for MAPK phosphorylation in response to known agonists.

 Results: Successful expression and coupling of both GPCRs was demonstrated as assessed by western blotting cellular extracts with IBA custom made specific rabbit polyclonal antibodies and by showing a >10-fold increase in phospho-MAPK in response to known agonists.

Project Completion: 3 months