Construction of Human Antibody Gene Libraries and Selection of Antibodies by Phage Display

  • André Frenzel
  • Jonas Kügler
  • Sonja Wilke
  • Thomas Schirrmann
  • Michael Hust
Part of the Methods in Molecular Biology book series (MIMB, volume 1060)


Antibody phage display is the most commonly used in vitro selection technology and has yielded thousands of useful antibodies for research, diagnostics, and therapy.

The prerequisite for successful generation and development of human recombinant antibodies using phage display is the construction of a high-quality antibody gene library. Here, we describe the methods for the construction of human immune and naive scFv gene libraries.

The success also depends on the panning strategy for the selection of binders from these libraries. In this article, we describe a panning strategy that is high-throughput compatible and allows parallel selection in microtiter plates.

Key words

Antibody Phage display scFv Library Naive library Immune library In vitro selection Panning 



We thank Stefan Dübel for discussion and corrections on the manuscript. We gratefully acknowledge the financial support by the FP7 collaborative projects AffinityProteome (contract 222635) and AFFINOMICS (contract 241481). This review is an updated and revised version of [54, 79] and the professorial dissertation (Habilitation) of Michael Hust.


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Copyright information

© Springer Science+Business Media, LLC 2014

Authors and Affiliations

  • André Frenzel
    • 1
  • Jonas Kügler
    • 2
  • Sonja Wilke
    • 2
  • Thomas Schirrmann
    • 3
  • Michael Hust
    • 3
  1. 1.Abteilung Biotechnologie Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und BioinformatikBraunschweigGermany
  2. 2.mAb-Factory GmbH, mAb-factory GmbHBraunschweigGermany
  3. 3.Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Abteilung BiotechnologieBraunschweigGermany

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