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Ligand Engineering via Yeast Surface Display and Adherent Cell Panning

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Genotype Phenotype Coupling

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

Abstract

High-throughput ligand discovery and evolution—via genotype-phenotype linkage strategies—empower molecularly targeted therapy, diagnostics, and fundamental science. Maintaining high-quality target antigen in these selections, particularly for membrane targets, is often a technical challenge. Panning yeast-displayed ligand libraries on intact mammalian cells expressing the molecular target has emerged as an effective strategy. Herein we describe the techniques used to select target-binding ligands via this approach including the use of target-negative cells to deplete non-specific binders and avidity reduction to preferentially select high-affinity ligands.

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Acknowledgments

This chapter describes work funded by the American Cancer Society (130418-RSG-17-110-01-TBG to B.J.H.), the National Institutes of Health (R01 EB023339 to B.J.H.), and the California Tobacco-Related Disease Research Grants Program Office of the University of California (28FT-0072 to L.A.S.).

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Author notes

  1. Lawrence A. Stern and Patrick S. Lown contributed equally to this work.

Authors and Affiliations

  1. Department of Hematology and Hematopoietic Cell Transplantation, Beckman Research Institute of the City of Hope, Duarte, CA, USA

    Lawrence A. Stern

  2. Department of Chemical Engineering and Materials Science, University of Minnesota—Twin Cities, Minneapolis, MN, USA

    Patrick S. Lown & Benjamin J. Hackel

Authors
  1. Lawrence A. Stern
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  2. Patrick S. Lown
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  3. Benjamin J. Hackel
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Corresponding author

Correspondence to Benjamin J. Hackel .

Editor information

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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Stern, L.A., Lown, P.S., Hackel, B.J. (2020). Ligand Engineering via Yeast Surface Display and Adherent Cell Panning. 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_17

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  • DOI: https://doi.org/10.1007/978-1-4939-9853-1_17

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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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