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Thioredoxin-Displayed Multipeptide Immunogens

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Book cover Peptide Antibodies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1348))

Abstract

Fusion to carrier proteins is an effective strategy for stabilizing and providing immunogenicity to peptide epitopes. This is commonly achieved by cross-linking of chemically synthesized peptides to carrier proteins. An alternative approach is internal grafting of selected peptide epitopes to a scaffold protein via double stranded-oligonucleotide insertion or gene synthesis, followed by recombinant expression of the resulting chimeric polypeptide. The scaffold protein should confer immunogenicity to the stabilized and structurally constrained peptide, but also afford easy production of the antigen in recombinant form. A macromolecular scaffold that meets the above criteria is the redox protein thioredoxin, especially bacterial thioredoxin. Here we describe our current methodology for internal grafting of selected peptide epitopes to thioredoxin as tandemly arranged multipeptide repeats (“Thioredoxin Displayed Multipeptide Immunogens”), bacterial expression and purification of the recombinant thioredoxin–multipeptide fusion proteins and their use as antigens for the production of anti-peptide antibodies for prophylactic vaccine as well as diagnostic purposes.

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Acknowledgments

This work was supported by grants from the Italian Association for Cancer Research (AIRC, grant IG 12956) and from the Regione Emilia Romagna, Ricerca Regione-Università 2010–2012, Strategic Programme “A tailored approach to the immune-monitoring and clinical management of viral and autoimmune diseases” to S.O. We thank Arturo Roberto Viscomi (GSK, San Polo di Torrile, Parma, Italy) for precious and highly competent help at an early stage of this work. E.C. was supported by a postdoctoral fellowship from AIRC.

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Authors and Affiliations

  1. Biochemistry and Molecular Biology Unit, Department of Life Sciences, University of Parma, Parma, Italy

    Angelo Bolchi, Elena Canali, Andrea Santoni, Gloria Spagnoli & Simone Ottonello

  2. German Cancer Research Center, Heidelberg, Germany

    Daniele Viarisio & Martin Müller

  3. Infections and Cancer Biology Group, International Agency for Research on Cancer-World Health Organization, Lyon, France

    Rosita Accardi & Massimo Tommasino

  4. Dipartimento di Bioscienze, Università di Parma, Parco Area delle Scienze 23/A, Parma, 43124, Italy

    Simone Ottonello

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  1. Angelo Bolchi
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  2. Elena Canali
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  3. Andrea Santoni
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  4. Gloria Spagnoli
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  5. Daniele Viarisio
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  6. Rosita Accardi
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  7. Massimo Tommasino
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  8. Martin Müller
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  9. Simone Ottonello
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Correspondence to Simone Ottonello .

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  1. Statens Serum Institut, Copenhagen, Denmark

    Gunnar Houen

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Bolchi, A. et al. (2015). Thioredoxin-Displayed Multipeptide Immunogens. 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_14

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  • DOI: https://doi.org/10.1007/978-1-4939-2999-3_14

  • 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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