Date of Award
A simple, reliable method is described for the routine measurement of non-steroidal anti-inflammatory drugs (NSAIDs) as inhibitors of prostaglandin synthase. Appropriate concentrations of inhibitor are incubated with ram seminal vesical prostaglandin synthase, sodium arachidonate and reduced glutathione. Prostaglandin E2 (PGE2) production is then quantitated by and enzyme linked immunosorbant assay (ELISA), in which anti-PGE2 antibody is utilized in a binding competition between test sample and adsorbed conjugate of PGE2-BSA. The antibody which remains bound to the solid phase is quantitatively determined colorimetrically by incubation with horse radish peroxidase labeled goat anti-rabbit IgG followed by incubation ·with N,N,N',N',-tetramethylbenzidine.
The relative inhibitory potency of Indomethacin was determined by comparison of PGE2 content in sample incubations, with that of controls containing no Indomethacin and was shown to inhibit prostaglandin synthase.
The combination of enzyme-inhibitor incubation assay with detection by ELISA permits the testing of other NSAID compounds for their ability to inhibit prostaglandin synthase activity expediently, and in a financially feasible manner. Furthermore, the use of an ELISA system eliminates radioactive and toxic chemical waste genera ted by radioimmu no assay methods.
Scott, Patricia, "A PROSTAGLANDIN SYNTHASE INHIBITION ASSAY WITH DETECTION BY ELISA" (1990). Theses, Dissertations & Honors Papers. 319.