Epitope Mapping via Phage Display from Single-Gene Libraries

  • Viola Fühner
  • Philip Alexander Heine
  • Kilian Johannes Carl Zilkens
  • Doris Meier
  • Kristian Daniel Ralph Roth
  • Gustavo Marçal Schmidt Garcia Moreira
  • Michael HustEmail author
  • Giulio Russo
Part of the Methods in Molecular Biology book series (MIMB, volume 1904)


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.

Key words

Phage display Epitope mapping Antigen fragments Protein fragments Panning 



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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Viola Fühner
    • 1
  • Philip Alexander Heine
    • 1
  • Kilian Johannes Carl Zilkens
    • 2
  • Doris Meier
    • 1
  • Kristian Daniel Ralph Roth
    • 1
  • Gustavo Marçal Schmidt Garcia Moreira
    • 1
  • Michael Hust
    • 1
    Email author
  • Giulio Russo
    • 1
  1. 1.Abteilung Biotechnologie, Institut für Biochemie, Biotechnologie und BioinformatikTechnische Universität BraunschweigBraunschweigGermany
  2. 2.YUMAB GmbHBraunschweigGermany

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