title: Detection of prion epitopes on PrP and PrP of transmissible spongiform encephalopathies using specific monoclonal antibodies to PrP.
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).
authors.affiliation: Australian Red Cross Blood Service-Endeavour, Sydney, New South Wales, Australia. fyuan@arcbs.redcross.org.au
authors.forename: Fang F
authors.initials: FF
authors.lastname: Yuan
authors.forename: Sandra
authors.initials: S
authors.lastname: Biffin
authors.forename: Marcus W
authors.initials: MW
authors.lastname: Brazier
authors.forename: Monica
authors.initials: M
authors.lastname: Suarez
authors.forename: Roberto
authors.initials: R
authors.lastname: Cappai
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Hill
authors.forename: Steven J
authors.initials: SJ
authors.lastname: Collins
authors.forename: John S
authors.initials: JS
authors.lastname: Sullivan
authors.forename: Deborah
authors.initials: D
authors.lastname: Middleton
authors.forename: Gerd
authors.initials: G
authors.lastname: Multhaup
authors.forename: Andrew F
authors.initials: AF
authors.lastname: Geczy
authors.forename: Colin L
authors.initials: CL
authors.lastname: Masters
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D018621:PrPC Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D018620:PrPSc Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D017096:Prion Diseases / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; D018621:PrPC Proteins; D018620:PrPSc Proteins