Monoclonal antibodies against bovine beta-casein: Production and epitope characterization

Twenty-one murine monoclonal antibodies (MAbs) were raised against bovine b eta -casein (beta -CN). They were discriminated by: (1) their specificity i n ACP-ELISA towards beta -CN fragments and beta -CN from buffalo, ewe, goat , human and mare; and (2) their reactivity in BIAcore assays with bovine be ta -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 clu stered epitopes except one were found to be overlapping by BIAcore additivi ty assay. Epitopes were delineated at position 4-8 and 178-183. One-quarter of the residues at both termini shared equally similar to 72% of the epito pes. No MAb was bovine-specific. A single MAb discriminated bovine whole ca sein from that of ewe and goat. Epitopes were included in each of the plasm in-released proteose peptone and gamma -CN peptides, and the panel of MAbs may thus provide probes suitable for their detection.