| Technologies Implemented at Berlex for Screening GPCRs Over the Past 10 Years |
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Presentation: 2B02 Session: Comparing HTS TechnologiesSofia Ribeiro, April Holbrook, Paymann Harirchian, Steve Jones, Lan Trinh, Laura Dunning, Lorraine Kent, Meina Liang, Mary Rosser, Kieu Chu and Michael Snider, Berlex Biosciences Presenting Author: Sofia Ribeiro, Berlex Biosciences - USA Over the past 10 years at Berlex, 10 GPCR projects have been implemented leading to the identification of small molecules GPCR antagonists that achieved clinical development. These successes validate the technologies used for HTS: radioligand receptor binding screens. Considerable effort was dedicated to further characterization of hits by secondary assays: calcium flux, cAMP, microphysiometry, platelet aggregation and chemotaxis. These various assays have been performed in whole cells naturally expressing the receptor of interest, or transfected with and overexpressing the receptor human whole blood or human washed platelets. Specific results in these projects have taught us that not all GPCRs are created equal. For example, one GPCR target was not amenable to HTS using receptor binding. We have identified both antagonists and agonists by primary receptor binding assays as well as by functional assays. Examples will be given of successful and unsuccessful assay development and lessons learned in the process. |