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Epitope Mapping Using Yeast Display and Next Generation Sequencing

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Epitope Mapping Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1785))

Abstract

Monoclonal antibodies are the largest class of therapeutic proteins due in part to their ability to bind an antigen with a high degree of affinity and specificity. A precise determination of their epitope is important for gaining insights into their therapeutic mechanism of action and to help differentiate antibodies that bind the same antigen. Here, we describe a method to precisely and efficiently map the epitopes of multiple antibodies in parallel over the course of just several weeks. This approach is based on a combination of rational library design, yeast surface display, and next generation DNA sequencing and provides quantitative insights into the epitope residues most critical for the antibody-antigen interaction. As an example, we will use this method to map the epitopes of several antibodies that neutralize alpha toxin from Staphylococcus aureus.

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Author information

Author notes

  1. Davide Foletti & Steven Pitts

    Present address: 23andMe Inc., South San Francisco, CA, USA

  2. Wai Ling Cheung

    Present address: Princeton University, Princeton, NJ, USA

  3. Jan Berka

    Present address: Roche Sequencing Solutions, Pleasanton, CA, USA

  4. Wenwu Zhai

    Present address: NGM Biopharmaceuticals Inc., South San Francisco, CA, USA

  5. Pavel Strop & Arvind Rajpal

    Present address: Bristol-Myers Squibb Inc., Redwood City, CA, USA

  6. Jaume Pons

    Present address: Alexo Therapeutics Inc., South San Francisco, CA, USA

Authors and Affiliations

  1. Rinat, Pfizer Inc., South San Francisco, CA, USA

    Thomas Van Blarcom, Andrea Rossi, Davide Foletti, Purnima Sundar, Steven Pitts, Zea Melton, Dilduz Telman, Lora Zhao, Wai Ling Cheung, Jan Berka, Wenwu Zhai, Pavel Strop, Jaume Pons, Arvind Rajpal & Javier Chaparro-Riggers

Authors
  1. Thomas Van Blarcom
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  2. Andrea Rossi
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  3. Davide Foletti
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  4. Purnima Sundar
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  5. Steven Pitts
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  6. Zea Melton
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  7. Dilduz Telman
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  8. Lora Zhao
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  9. Wai Ling Cheung
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  10. Jan Berka
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  11. Wenwu Zhai
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  12. Pavel Strop
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  13. Jaume Pons
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  14. Arvind Rajpal
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  15. Javier Chaparro-Riggers
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Corresponding author

Correspondence to Thomas Van Blarcom .

Editor information

Editors and Affiliations

  1. KTH School of Engineering Sciences in Chemistry, Biotechnology and Health, Protein Technology, Stockholm, Sweden

    Johan Rockberg

  2. Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada

    Johan Nilvebrant

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Van Blarcom, T. et al. (2018). Epitope Mapping Using Yeast Display and Next Generation Sequencing. In: Rockberg, J., Nilvebrant, J. (eds) Epitope Mapping Protocols. Methods in Molecular Biology, vol 1785. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7841-0_7

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  • DOI: https://doi.org/10.1007/978-1-4939-7841-0_7

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7839-7

  • Online ISBN: 978-1-4939-7841-0

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