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

✍ Scribed by Scott P. Ager; G.Michael Hass

• Publisher: Elsevier Science
• Year: 1977
• Tongue: English
• Weight: 685 KB
• Volume: 83
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(77)90537-1

✦ Synopsis

A two-step purification scheme based on affmity 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 trypsintreated extracts of pancreatic acetone powder to immobilized carboxypeptidase inhibitor from potatoes results in the specific retention of the target exopeptidases. After elution 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 c-amino-n-caproyl-o-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.