Immunological identification of milk-clotting enzymes

Abstract

SUMMARYThe 6 most widely used milk-clotting enzymes, i.e. chymosin, bovine and porcine pepsins and proteinases from Mucor miehei, M. pusillus and Endothia parasitica, have been purified and used to prepare rabbit antisera against each of them. The antisera were adsorbed with appropriate cross-linked antigens to remove non-specific antibodies. Monospecific antisera thus obtained were used to identify enzymes eventually added to calf or bovine rennets, using the double radial immunodiffusion technique. The threshold of sensitivity was c. 1%, expressed as clotting activity. This qualitative method complements the quantitative procedure recently proposed for determining chymosin and bovine pepsin A in commercial extracts of bovine veils.