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A Streamlined Approach for the Construction of Large Yeast Surface Display Fab Antibody Libraries

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Antibody Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1827))

Abstract

Yeast surface display is a versatile platform technology for antibody discovery. Nevertheless, the construction of antibody Fab libraries typically is a tedious multistep process that involves the generation of heavy chain as well as light chain display plasmids in different haploid yeast strains followed by yeast mating. Here, we present a focused one-step Golden Gate cloning approach for the generation of yeast surface display Fab libraries that allows for simultaneous introduction of heavy-chain and light-chain variable regions into one single display vector. Thereby, the overall time as well as the materials needed for library generation can be reduced significantly.

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Author notes

  1. Simon Krah and Julius Grzeschik contributed equally to this work.

Authors and Affiliations

  1. Institute for Organic Chemistry and Biochemistry, Technische Universität Darmstadt, Darmstadt, Germany

    Simon Krah, Julius Grzeschik, Simon Rosowski, Harald Kolmar & Stefan Zielonka

  2. Protein Engineering and Antibody Technologies, Merck KGaA, Darmstadt, Germany

    Simon Krah, Ramona Gaa, Iris Willenbuecher, Deniz Demir, Lars Toleikis, Stefan Becker & Stefan Zielonka

Authors
  1. Simon Krah
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  2. Julius Grzeschik
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  3. Simon Rosowski
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  4. Ramona Gaa
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  5. Iris Willenbuecher
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  6. Deniz Demir
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  7. Lars Toleikis
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  8. Harald Kolmar
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  9. Stefan Becker
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  10. Stefan Zielonka
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Corresponding author

Correspondence to Stefan Zielonka .

Editor information

Editors and Affiliations

  1. Immunology Division, Garvan Institute of Medical Research, Darlinghurst, NSW, Australia

    Damien Nevoltris

  2. Institut Paoli-Calmettes, CRCM, Aix Marseille University, CNRS, INSERM, Marseille, France

    Patrick Chames

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Krah, S. et al. (2018). A Streamlined Approach for the Construction of Large Yeast Surface Display Fab Antibody Libraries. In: Nevoltris, D., Chames, P. (eds) Antibody Engineering. Methods in Molecular Biology, vol 1827. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8648-4_8

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  • DOI: https://doi.org/10.1007/978-1-4939-8648-4_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8647-7

  • Online ISBN: 978-1-4939-8648-4

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