Prion-Specific Antibodies Produced in Wild-Type Mice

Peter M. H. Heegaard, Ann-Louise Bergström, Heidi Gertz Andersen, Henriette Cordes Hvass

    Research output: Contribution to journalJournal articleResearchpeer-review

    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.
    Original languageEnglish
    JournalMethods in Molecular Biology
    Volume1348
    Pages (from-to)285-301
    Number of pages17
    ISSN1064-3745
    DOIs
    Publication statusPublished - 2015

    Keywords

    • Monoclonal antibodies
    • Peptide-carrier protein conjugates
    • Peptide-specific antibodies
    • Prion
    • Synthetic peptide

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