NR AIBJ
AU Matsuda,H.; Mitsuda,H.; Nakamura,N.; Furusawa,S.; Mohri,S.; Kitamoto,T.
TI A chicken monoclonal antibody with specificity for the N-terminal of human prion protein
QU FEMS Immunology and Medical Microbiology 1999 Mar; 23(3): 189-94
PT journal article
AB 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-Sträussler 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.
MH Amino Acid Sequence; Animal; Antibodies, Monoclonal/*immunology; Antibody Specificity; Blotting, Western; Cattle; Cell Line; Chickens; Hamsters; Human; Immunohistochemistry; Mice; Molecular Sequence Data; Prions/*immunology; Sheep; Support, Non-U.S. Gov't
AD Department of Immunobiology, Faculty of Applied Biological Science, Hiroshima University, Japan. hmatsu@ipc.hiroshima-u.ac.jp
SP englisch
PO Niederlande