title: Prion-Specific Antibodies Produced in Wild-Type Mice.
abstract: Peptide-specific antibodies produced against synthetic peptides are of high value in probing protein structure and function, especially when working with challenging proteins, including not readily available, non-immunogenic, toxic, and/or pathogenic proteins. Here, we present a straightforward method for production of mouse monoclonal antibodies (MAbs) against peptides representing two sites of interest in the bovine prion protein (boPrP), the causative agent of bovine spongiform encephalopathy ("mad cow disease") and new variant Creutzfeldt-Jakob's disease (CJD) in humans, as well as a thorough characterization of their reactivity with a range of normal and pathogenic (misfolded) prion proteins. It is demonstrated that immunization of wild-type mice with ovalbumin-conjugated peptides formulated with Freund's adjuvant induces a good immune response, including high levels of specific anti-peptide antibodies, even against peptides very homologous to murine protein sequences. In general, using the strategies described here for selecting, synthesizing, and conjugating peptides and immunizing 4-5 mice with 2-3 different peptides, high-titered antibodies reacting with the target protein are routinely obtained with at least one of the peptides after three to four immunizations with incomplete Freund's adjuvant.
authors.affiliation: Innate Immunology Group, Section for Immunology and Vaccinology, National Veterinary Institute, Technical University of Denmark, Building B, Bülowsvej 27, 1870, Frederiksberg, Denmark. PMHH@vet.dtu.dk.
authors.forename: Peter M H
authors.initials: PM
authors.lastname: Heegaard
authors.affiliation: Innate Immunology Group, Section for Immunology and Vaccinology, National Veterinary Institute, Technical University of Denmark, Building B, Bülowsvej 27, 1870, Frederiksberg, Denmark.|Department Neurodegeneration, H. Lundbeck A/S, Ottiliavej 9, 2500, Valby, Denmark.
authors.forename: Ann-Louise
authors.initials: AL
authors.lastname: Bergström
authors.affiliation: Innate Immunology Group, Section for Immunology and Vaccinology, National Veterinary Institute, Technical University of Denmark, Building B, Bülowsvej 27, 1870, Frederiksberg, Denmark.
authors.forename: Heidi Gertz
authors.initials: HG
authors.lastname: Andersen
authors.affiliation: Innate Immunology Group, Section for Immunology and Vaccinology, National Veterinary Institute, Technical University of Denmark, Building B, Bülowsvej 27, 1870, Frederiksberg, Denmark.|Novo Nordisk A/S, Krogshøjvej 30, 2880, Bagsværd, Denmark.
authors.forename: Henriette
authors.initials: H
authors.lastname: Cordes
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology*; D002352:Carrier Proteins; D002417:Cattle; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D006825:Hybridomas / Q000276:immunology; D007114:Immunization; D051379:Mice; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011328:Prions / Q000235:genetics / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D000939:Epitopes; D010455:Peptides; D011328:Prions
keywords: Monoclonal antibodies; Peptide-carrier protein conjugates; Peptide-specific antibodies; Prion; Synthetic peptide