Purification of recombinant human COX-1 and COX-2.

Abstract

Human recombinant COX-1 and COX-2 was prepared in a purified form. The genes were cloned and expressed in insect cells, extracted by detergents, and purified by ion-exchange followed by size exclusion chromatography. Insect cells from 10 L fermentation yielded 2.3 mg of COX-1 with an overall yield of 75%, and 29 mg of COX-2 with an overall yield of 45%. Enzyme prepared in this manner was fully active and proved to be useful in biophysical studies including direct binding studies. The COX-2 provided material that was subsequently used in X-ray crystallography studies.