Monoclonal Antibodies against Bovine -Casein: Production and Epitope Characterization

Twenty-one murine monoclonal antibodies (MAbs) were raised against bovine -casein (-CN). They were discriminated by: (1) their specificity in ACP-ELISA towards -CN fragments and -CN from buffalo, ewe, goat, human and mare; and (2) their reactivity in BIAcore assays with bovine -CN. The antigen sequences were compared to delineate epitopes. The MAbs recognized 14 distinct epitopes clustered in six discrete determinants (4–8, 14–24, 33–48, 49–91, 178–183, 184–209). Within the determinants, all clustered epitopes except one were found to be overlapping by BIAcore additivity assay. Epitopes were delineated at position 4–8 and 178–183. One-quarter of the residues at both termini shared equally ~ 72% of the epitopes. No MAb was bovine-specific. A single MAb discriminated bovine whole casein from that of ewe and goat. Epitopes were included in each of the plasmin-released proteose peptone and -CN peptides, and the panel of MAbs may thus provide probes suitable for their detection.