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Fluorescence polarization based assay suitable for screening for H-Prostaglandin D Synthase inhibitors

Carl Peters, BMG LABTECH, Cary, NC, 07/2015

Prostaglandins (PG) are tissue hormones that are involved in many physiological processes. One of the most well-known prostaglandins is PGD2 which, together with other mediators, causes bronchial contraction. It has been shown that PGD2 concentration exceeds normal state levels in asthma patients. For this and other reasons, researchers are searching for ways to decrease PGD2 concentration. One possibility is to look for inhibitors that block the last step of the PGD2 biosynthesis.

Cayman Chemical developed a fluorescence polarization (FP) based competition assay that provides the possibility to screen for compounds that are able to inhibit the hematopoietic prostaglandin D synthase. A fluorescein labeled inhibitor is added to interact with the synthase, resulting in a high FP value. Test compounds compete with the labeled inhibitor for the synthase binding sites. Successful compounds can be detected via a low FP value. This assay is a great advantage compared to the traditional multi-step assay which involves the handling of the highly unstable PGD2-precursor.

The application note shows inhibitor dose response curves obtained on the PHERAstar FS and CLARIOstar microplate readers. Excellent Z' values show the reliability of the assay and of the instrumentation.