Novel epitopes identified by anti-PrP monoclonal antibodies produced following immunization of Prnp0/0 Balb/cJ mice with purified scrapie prions

Hybridoma (Larchmt). 2012 Oct;31(5):314-24. doi: 10.1089/hyb.2012.0022.

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).

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / chemistry*
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / isolation & purification
  • Antibody Specificity
  • Binding Sites, Antibody
  • Blotting, Western
  • Brain Chemistry
  • Cricetinae
  • Enzyme-Linked Immunosorbent Assay
  • Epitope Mapping*
  • Epitopes / chemistry*
  • Epitopes / immunology
  • Humans
  • Immunization
  • Membrane Microdomains / chemistry
  • Mesocricetus
  • Mice
  • Mice, Transgenic
  • Models, Molecular
  • Molecular Sequence Data
  • Peptides / chemical synthesis*
  • Peptides / immunology
  • PrPSc Proteins / administration & dosage
  • PrPSc Proteins / chemistry*
  • PrPSc Proteins / immunology
  • PrPSc Proteins / isolation & purification
  • Protein Structure, Secondary

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • Peptides
  • PrPSc Proteins