A PROSTAGLANDIN SYNTHASE INHIBITION ASSAY WITH DETECTION BY ELISA

Author

Patricia Scott, Longwood University

Date of Award

4-25-1990

Degree Type

Honors Paper

Department

Chemistry

Abstract

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.

Recommended Citation

Scott, Patricia, "A PROSTAGLANDIN SYNTHASE INHIBITION ASSAY WITH DETECTION BY ELISA" (1990). Theses & Honors Papers. 319.
https://digitalcommons.longwood.edu/etd/319