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Epitope Mapping via Phage Display from Single-Gene Libraries

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Human Monoclonal Antibodies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1904))

Abstract

Antibodies are widely used in a large variety of research applications, for diagnostics and therapy of numerous diseases, primarily cancer and autoimmune diseases. Antibodies are binding specifically to target structures (antigens). The antigen-binding properties are not only dependent on the antibody sequence, but also on the discrete antigen region recognized by the antibody (epitope). Knowing the epitope is valuable information for the improvement of diagnostic assays or therapeutic antibodies, as well as to understand the immune response of a vaccine. While huge progress has been made in the pipelines for the generation and functional characterization of antibodies, the available technologies for epitope mapping are still lacking effectiveness in terms of time and effort. Also, no technique available offers the absolute guarantee of succeeding. Thus, research to develop and improve epitope mapping techniques is still an active field. Phage display from random peptide libraries or single-gene libraries are currently among the most exploited methods for epitope mapping. The first is based on the generation of mimotopes and it is fastened to the need of high-throughput sequencing and complex bioinformatic analysis. The second provides original epitope sequences without requiring complex analysis or expensive techniques, but depends on further investigation to define the functional amino acids within the epitope. In this book chapter, we describe how to perform epitope mapping by antigen fragment phage display from single-gene antigen libraries and how to construct these types of libraries. Thus, we also provide figures and analysis to demonstrate the actual potential of this technique and to prove the necessity of certain procedural steps.

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Acknowledgments

We thank CNPq for providing the scholarship of GMSGM (process 204693/2014-4). V.F. was funded by the Federal State of Lower Saxony, Niedersächsisches Vorab (VWZN2889/3215/3266). This chapter is a completely revised and updated version of [46].

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Authors and Affiliations

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

    Viola Fühner, Philip Alexander Heine, Doris Meier, Kristian Daniel Ralph Roth, Gustavo Marçal Schmidt Garcia Moreira, Michael Hust & Giulio Russo

  2. YUMAB GmbH, Braunschweig, Germany

    Kilian Johannes Carl Zilkens

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  1. Viola Fühner
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  2. Philip Alexander Heine
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  3. Kilian Johannes Carl Zilkens
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  4. Doris Meier
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  5. Kristian Daniel Ralph Roth
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  6. Gustavo Marçal Schmidt Garcia Moreira
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  7. Michael Hust
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  8. Giulio Russo
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Correspondence to Michael Hust .

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  1. Department of Pathology, The Hebrew University, Hadassah Medical School, Jerusalem, Israel

    Michael Steinitz

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Fühner, V. et al. (2019). Epitope Mapping via Phage Display from Single-Gene Libraries. In: Steinitz, M. (eds) Human Monoclonal Antibodies. Methods in Molecular Biology, vol 1904. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8958-4_17

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  • DOI: https://doi.org/10.1007/978-1-4939-8958-4_17

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  • Publisher Name: Humana Press, New York, NY

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  • Online ISBN: 978-1-4939-8958-4

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