A chicken monoclonal antibody with specificity for the N-terminal of human prion protein

Chickens were immunized with human prion protein (PrP) peptide H25 (amino acid residues 25–49) coupled to keyhole limpet hemocyanin. From a fusion experiment using the chicken fusion partner cell line MuH1 and immune spleen cells, one mAb, HUC2–13, was generated which reacted with the peptide. HUC2–13 was specific for a pentapeptide (RPKPG) of the N-terminal of the peptide H25. In Western blotting analysis, the mAb reacted with PrP materials from a human Creutzfeldt—Jakob disease (CJD) case and the membrane fraction from normal murine brain, but not with the same materials pretreated with proteinase K. When compared with the HUC2–13 and the conventional mouse mAb 3F4, the background stainings using the HUC2–13 were minimal. In immunohistochemistry, the HUC2–13 stained positively with kuru plaques in brain sections from patients with Gerstmann—Straussler syndrome (GSS), and also reacted with synaptic structures of the CJD patients. However, any immunolabelings using the HUC2–13 were not observed in the section from a patient with amyotrophic lateral sclerosis (ALS) as CJD-negative control. These results indicate that the mAb HUC2–13 is a suitable tool for immunological and diagnostic analyses of prion disease in humans and other mammals.