Immunochemical, chromatographic, and milk-clotting activity measurements for quantification of milk-clotting enzymes in bovine rennets

Abstract

Summary: Monospecific rabbit antibodies were produced against bovine chymosin (rennin) and bovine pepsin A and used for quantitative measurements of these enzymes by rocket immunoelectrophoresis. Bovine pepsin B was identified and measured quantitatively in tandem crossed immunoelectrophoresis, using a polyspecific antibody preparation. Chromatographically purified chymosin B and bovine pepsins A and B were used as standards. The contents of chymosin and bovine pepsin A were also determined by 2 forms of DEAE chromatography, and by an assay utilizing the ratio of milk-clotting activity at pH 6·0 and 6·5. Results obtained by measurements on 3 bovine rennets (calf rennet, adult bovine rennet, and a mixture of such types) by the different methods of analysis were in good agreement. The advantages of the immunochemical methods are direct and simultaneous identification and quantitation of enzymes.