Affinity chromatography of pancreatic carboxypeptidases using a carboxypeptidase inhibitor from potatoes as ligand

A two-step purification scheme based on affinity chromatography has been developed for bovine and porcine carboxypeptidases A and B. Enzyme preparations of high purity are achieved from extracts of pancreatic acetone powder in less than 1 day by this procedure. The application of trypsin-treated extracts of pancreatic acetone powder to immobilized carboxypeptidase inhibitor from potatoes results in the specific retention of the target exopeptidases. After clution of the bound carboxypeptidases A and B at high pH, the resultant mixture of the two enzymes is resolved by chromatography on a column of ϵ-amino- n-caproyl- d-arginine-Sepharose 4B. Overall yields of 70–80% of the purified enzymes have been obtained with no cross-contamination of detectable tryptic or chymotryptic activities of the preparations.