Chicken monoclonal antibodies against synthetic bovine prion protein peptide

Chicken monoclonal antibodies (mAbs) were developed against bovine prion protein (PrP) peptide. Chickens immunized with bovine PrP peptide B204 (amino acid residues 204-220) coupled to keyhole limpet hemocyanin produced specific antibodies to the peptide as determined by an enzyme-linked immunosorbent assay (ELISA) using the B204 peptide coupled to ovalbumin as target antigen. From a fusion experiment using the chicken fusion partner cell line MuH1 and immune spleen cells, 19 mAbs reactive with B204 were generated. These mAbs were subdivided into five groups based on competitive ELISA using B204 and four 10-amino acid peptides. These five groups included all combinations expected based on comparison of amino acid sequences among the five species, bovine, mouse, human, sheep and hamster, examined. These results indicate that the chicken mAb system is a suitable technique for immunological analysis of PrP in mammals.