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Phage Display pp 477–495Cite as

ORFeome Phage Display

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1701))

Abstract

ORFeome phage display allows the efficient functional screening of entire proteomes or even metaproteomes to identify immunogenic proteins. For this purpose, randomly fragmented, whole genomes or metagenomes are cloned into a phage-display vector allowing positive selection for open reading frames (ORF) to improve the library quality. These libraries display all possible proteins encoded by a pathogen or a microbiome on the phage surface. Consequently, immunogenic proteins can be selected from these libraries using disease-related immunoglobulins from patient serum. ORFeome phage display in particular allows the identification of immunogenic proteins that are only expressed in the host-pathogen interaction but not in cultivation, as well as the detection of very low expressed and very small immunogens and immunogenic proteins of non-cultivable organisms. The identified immunogenic proteins are potential biomarkers for the development of diagnostic assays or vaccines. These articles will give an introduction to ORFeome phage-display technology and give detailed protocols to identify immunogenic proteins by phage display.

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Acknowledgments

The introduction of this article contains updated and extended parts of the German language article Zantow et al. [23] and the dissertation of Jonas Zantow, TU Braunschweig.

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Authors and Affiliations

  1. Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung Biotechnologie, Technische Universität Braunschweig, Spielmannstr. 7, 38106, Braunschweig, Germany

    Jonas Zantow & Stefan Dübel

  2. Abteilung Biotechnologie, Institut für Biochemie, Biotechnologie und Bioinformatik, Technische Universität Braunschweig, Braunschweig, Germany

    Gustavo Marçal Schmidt Garcia Moreira & Michael Hust

Authors
  1. Jonas Zantow
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  2. Gustavo Marçal Schmidt Garcia Moreira
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  3. Stefan Dübel
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  4. Michael Hust
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Corresponding author

Correspondence to Michael Hust .

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Editors and Affiliations

  1. Technische Universität Braunschweig, Braunschweig, Germany

    Michael Hust

  2. Institute for Research in Molecular Medicine, Universiti Sains Malaysia, Minden, Penang, Malaysia

    Theam Soon Lim

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Zantow, J., Moreira, G.M.S.G., Dübel, S., Hust, M. (2018). ORFeome Phage Display. In: Hust, M., Lim, T. (eds) Phage Display. Methods in Molecular Biology, vol 1701. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7447-4_27

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  • DOI: https://doi.org/10.1007/978-1-4939-7447-4_27

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7446-7

  • Online ISBN: 978-1-4939-7447-4

  • eBook Packages: Springer Protocols

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