The isolation of chymotrypsin-like enzymes by affinity chromatography using sepharose-4-phenylbutylamine.

Abstract

Affinity chromatography of chymotrypsin-like proteases on a column of Sepharose–4-phenylbutylamine (PBA) has been developed. Sepharose–PBA (Sepharose–NH∙[CH2]4∙C6H5) has been shown to selectively adsorb chymotrypsin α and B from weakly alkaline solutions and to allow to pass through unretarded porcine trypsin, bovine trypsinogen, and chymotrypsin α modified with active-site-directed irreversible inhibitors. Chymotrypsinogen A and bovine trypsin were only slightly retarded whereas a preparation of subtilisin was markedly retarded and separated into two distinct peaks. Sepharose–PBA has been utilized successfully for the selective isolation of chymotryps in-like proteases from extracts of moose pancreas (Alces alces).