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.
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Acknowledgments
We would like to thank Regina Lund, Karin Larsen, Panchale Olsen, Annie Ravn Pedersen, and Anne-Marie Petersen for their excellent technical assistance.
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Heegaard, P.M.H., Bergström, AL., Andersen, H.G., Cordes, H. (2015). Prion-Specific Antibodies Produced in Wild-Type Mice. In: Houen, G. (eds) Peptide Antibodies. Methods in Molecular Biology, vol 1348. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2999-3_25
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DOI: https://doi.org/10.1007/978-1-4939-2999-3_25
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2998-6
Online ISBN: 978-1-4939-2999-3
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