Abstract
Hog pancreatic kallikrein A and B were separated by DEAE-Sephadex A-50 chromatography and purified by preparative high performance liquid chromatography (TSK GEL G2000 SW column). Kallkrein A and B were homogeneous in SDS-polyacrylamide gel electrophoresis. Antisera were elicited in rabbits by immunization of these purified antigens. The precipitin lines formed during diffusion of kallikrein A and B against the antisera, using double diffusion method, were completely comfluent and relative cross reactivity of respective antisera were quite similar. The Fab'fragments were conjugated with β-D-galactosidase using N, N-o-phenylene-dimaleimide as a coupling reagent. Using this antibody-enzyme complex, it was possible to determine as little as 1 ng/ml of kallikrein by the sandwich enzyme immunoassay. The results obtained with various samples of kallikrein (K-1000, K-200, K-100 and K-25) were in accord with the activities of biological assay.
