Abstract

Conventional approaches to purifying most connective tissue proteoglycans (PGs) generally involve treatments that consequently alter the binding properties of the purified PG (e.g., exposure to strong detergents or harsh denaturants such as guanidinium). Herein we describe a protocol that we use to isolate human decorin in which the PG remains functional in its binding to various ligands (1,2). This method utilizes a eukaryotic expression system that produces high quantities of recombinant human decorin as a soluble component of the transfected cell culture medium together with a combination of conventional ion-exchange and hydrophobic-interaction chromatography. Here we provide detailed instructions for (1) the production of conditioned medium containing secreted recombinant human decorin, (2) the initial ion-exchange chromatography step using DEAE-Sepharose, and (3) the final hydrophobic interaction chromatography step.

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