title: Novel epitopes identified by anti-PrP monoclonal antibodies produced following immunization of Prnp0/0 Balb/cJ mice with purified scrapie prions.
abstract: Prions, or infectious proteins, cause a class of uniformly fatal neurodegenerative diseases. Prions are composed solely of an aberrantly folded isoform (PrP(Sc)) of a normal cellular protein (PrP(C)). Shared sequence identity of PrP(Sc) with PrP(C) has limited the detection sensitivity of immunochemical assays, as antibodies specific for the disease-causing PrP(Sc) isoform have not been developed. Here we report the generation of three new monoclonal antibodies (MAbs) to PrP, which were isolated following immunization of Prnp(0/0) Balb/cJ mice with highly purified PrP(Sc) isolated from brain lipid rafts. Epitope mapping using synthetic PrP peptides revealed that the three MAbs bind different epitopes of PrP. The DRM1-31 MAb has a conformational epitope at the proposed binding site for the putative prion conversion co-factor "protein X." The DRM1-60 MAb binds a single linear epitope localized to the β2-α2 loop region of PrP, whereas DRM2-118 binds an epitope that includes sequences within the octarepeat region and near the site of N-terminal truncation of PrP(Sc) by proteinase K. Our novel anti-PrP MAbs with defined PrP epitopes may be useful in deciphering the conformational conversion of PrP(C) into PrP(Sc).
authors.affiliation: USDA-Agriculture Research Service, Foodborne Contaminants Research Unit, Albany, California 94710, USA.
authors.forename: Larry H
authors.initials: LH
authors.lastname: Stanker
authors.forename: Miles C
authors.initials: MC
authors.lastname: Scotcher
authors.forename: Alice
authors.initials: A
authors.lastname: Lin
authors.forename: Jeffery
authors.initials: J
authors.lastname: McGarvey
authors.forename: Stanley B
authors.initials: SB
authors.lastname: Prusiner
authors.forename: Robert
authors.initials: R
authors.lastname: Hnasko
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D000918:Antibody Specificity; D001666:Binding Sites, Antibody; D015153:Blotting, Western; D001923:Brain Chemistry; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D000939:Epitopes / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D007114:Immunization; D021962:Membrane Microdomains / Q000737:chemistry; D008647:Mesocricetus; D051379:Mice; D008822:Mice, Transgenic; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000276:immunology; D018620:PrPSc Proteins / Q000008:administration & dosage / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D017433:Protein Structure, Secondary
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D010455:Peptides; D018620:PrPSc Proteins