Detection of prion epitopes on PrP and PrP of transmissible spongiform encephalopathies using specific monoclonal antibodies to PrP

Immunol Cell Biol. 2005 Dec;83(6):632-7. doi: 10.1111/j.1440-1711.2005.01384.x.

Abstract

Amino acid residues 90-120 of the prion protein (PrP) are likely to be critical for the conversion of PrP(c) to PrP(sc) in the transmissible spongiform encephalopathies. We raised 10 monoclonal antibodies against the 90-120 amino acid region, mapped the epitope specificity of these anti-PrP antibodies, and investigated the expression of epitopes recognized by the antibodies in both PrP(c) and PrP(sc). Four out of five of the anti-PrP antibodies raised in a prion knockout mouse immunized with the linear peptide of PrP90-120 could detect PrP(sc) in 'native' and denatured forms and PrP(c) in normal cells, as well as recognize epitopes within PrP93-112 residues. In contrast, the other six anti-PrP reagents, including five raised from the two knockout mice immunized with conformationally modified PrP90-120 peptide, could detect PrP(c) and recognize epitopes within PrP93-107 residues. Four of these reagents could also detect denatured PrP(sc) on western blots but not PrP(sc) plaques in brain tissue. The results indicate that residues PrP93-102 are exposed in PrP(c) but are buried upon conversion to the PrP(sc) isoform. Furthermore, PrP103-107 residues are partially buried in PrP(sc) while only the PrP107-112 epitope remains exposed, suggesting that the region PrP93-112 undergoes conformational changes during its conversion to PrP(sc).

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity
  • Epitopes / chemistry
  • Epitopes / genetics
  • Epitopes / immunology*
  • Humans
  • Immunohistochemistry
  • Mice
  • PrPC Proteins / chemistry
  • PrPC Proteins / genetics
  • PrPC Proteins / immunology*
  • PrPSc Proteins / chemistry
  • PrPSc Proteins / genetics
  • PrPSc Proteins / immunology*
  • Prion Diseases / genetics
  • Prion Diseases / immunology*
  • Prion Diseases / metabolism*

Substances

  • Antibodies, Monoclonal
  • Epitopes
  • PrPC Proteins
  • PrPSc Proteins