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

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Human Monoclonal Antibodies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1060))

Abstract

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.

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Acknowledgements

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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Authors and Affiliations

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

    André Frenzel

  2. mAb-Factory GmbH, mAb-factory GmbH, Braunschweig, Germany

    Jonas Kügler & Sonja Wilke

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

    Thomas Schirrmann & Michael Hust

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  1. André Frenzel
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  2. Jonas Kügler
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  1. Department of Pathology, The Lautenberg Center, IMRIC The Hebrew University Hadassah Medical School, Jerusalem, Israel

    Michael Steinitz

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Frenzel, A., Kügler, J., Wilke, S., Schirrmann, T., Hust, M. (2014). Construction of Human Antibody Gene Libraries and Selection of Antibodies by Phage Display. In: Steinitz, M. (eds) Human Monoclonal Antibodies. Methods in Molecular Biology, vol 1060. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-62703-586-6_12

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  • DOI: https://doi.org/10.1007/978-1-62703-586-6_12

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