title: Prion protein-specific antibodies that detect multiple TSE agents with high sensitivity.
abstract: This paper describes the generation, characterisation and potential applications of a panel of novel anti-prion protein monoclonal antibodies (mAbs). The mAbs were generated by immunising PRNP null mice, using a variety of regimes, with a truncated form of recombinant ovine prion protein spanning residues 94-233. Epitopes of specific antibodies were mapped using solid-phase Pepscan analysis and clustered to four distinct regions within the PrP molecule. We have demonstrated the utility of these antibodies by use of Western blotting and immunohistochemistry in tissues from a range of different species affected by transmissible spongiform encephalopathy (TSE). In comparative tests against extensively-used and widely-published, commercially available antibodies, similar or improved results can be obtained using these new mAbs, specifically in terms of sensitivity of detection. Since many of these antibodies recognise native PrPC, they could also be applied to a broad range of immunoassays such as flow cytometry, DELFIA analysis or immunoprecipitation. We are using these reagents to increase our understanding of TSE pathogenesis and for use in potential diagnostic screening assays.
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: Sandra
authors.initials: S
authors.lastname: McCutcheon
authors.affiliation: Central Veterinary Institute of Wageningen UR, Lelystad, The Netherlands.
authors.forename: Jan P M
authors.initials: JP
authors.lastname: Langeveld
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: Boon Chin
authors.initials: BC
authors.lastname: Tan
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: Andrew C
authors.initials: AC
authors.lastname: Gill
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: Christopher
authors.initials: C
authors.lastname: de Wolf
authors.affiliation: Animal Health and Veterinary Laboratories Agency, Lasswade Laboratory, Edinburgh, Scotland, United Kingdom.
authors.forename: Stuart
authors.initials: S
authors.lastname: Martin
authors.affiliation: Animal Health and Veterinary Laboratories Agency, Lasswade Laboratory, Edinburgh, Scotland, United Kingdom.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Gonzalez
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: James
authors.initials: J
authors.lastname: Alibhai
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: A Richard Alejo
authors.initials: AR
authors.lastname: Blanco
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: Lauren
authors.initials: L
authors.lastname: Campbell
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: Nora
authors.initials: N
authors.lastname: Hunter
authors.affiliation: Neurobiology Division, The Roslin Institute and Royal (Dick) School of Veterinary Sciences, Easter Bush, Edinburgh, Scotland, United Kingdom.
authors.forename: E Fiona
authors.initials: EF
authors.lastname: Houston
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D001120:Arginine / Q000235:genetics; D001665:Binding Sites; D003062:Codon / Q000235:genetics; D007132:Immunoglobulin Isotypes / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D008969:Molecular Sequence Data; D018620:PrPSc Proteins / Q000378:metabolism; D017096:Prion Diseases / Q000276:immunology*; D011328:Prions / Q000737:chemistry / Q000276:immunology*; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D016415:Sequence Alignment; D012756:Sheep
chemical_list: D000911:Antibodies, Monoclonal; D003062:Codon; D007132:Immunoglobulin Isotypes; D018620:PrPSc Proteins; D011328:Prions; D011994:Recombinant Proteins; D001120:Arginine; D011494:Protein Kinases