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Isolation of Antigen-Specific VHH Single-Domain Antibodies by Combining Animal Immunization with Yeast Surface Display

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Genotype Phenotype Coupling

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2070))

Abstract

In addition to conventional hetero-tetrameric antibodies, the adaptive immune repertoire of camelids comprises the so-called heavy chain-only antibodies devoid of light chains. Consequently, antigen binding is mediated solely by the variable domain of the heavy chain, referred to as VHH. In recent years, these single-domain moieties emerged as promising tools for biotechnological and biomedical applications. In this chapter, we describe the generation of VHH antibody yeast surface display libraries from immunized Alpacas and Lamas as well as the facile isolation of antigen-specific molecules in a convenient fluorescence-activated cell sorting (FACS)-based selection process.

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Acknowledgments

We like to thank Preclinics GmbH for collaborating on this project.

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Authors and Affiliations

  1. Protein Engineering and Antibody Technologies (PEAT), Merck KGaA, Darmstadt, Germany

    Lukas Roth, Simon Krah, Janina Klemm, Ralf Günther, Lars Toleikis, Stefan Becker & Stefan Zielonka

  2. Discovery Pharmacology, Merck KGaA, Darmstadt, Germany

    Michael Busch

Authors
  1. Lukas Roth
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  2. Simon Krah
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  3. Janina Klemm
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  4. Ralf Günther
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  5. Lars Toleikis
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  6. Michael Busch
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  7. Stefan Becker
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  8. Stefan Zielonka
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Corresponding author

Correspondence to Stefan Zielonka .

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Editors and Affiliations

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

    Stefan Zielonka

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

    Simon Krah

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Roth, L. et al. (2020). Isolation of Antigen-Specific VHH Single-Domain Antibodies by Combining Animal Immunization with Yeast Surface Display. In: Zielonka, S., Krah, S. (eds) Genotype Phenotype Coupling. Methods in Molecular Biology, vol 2070. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9853-1_10

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  • DOI: https://doi.org/10.1007/978-1-4939-9853-1_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9852-4

  • Online ISBN: 978-1-4939-9853-1

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