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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 insight 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 insight 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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Epitope Mapping Using Yeast Display and Next Generation Sequencing

  • 实验方案

  • 发表日期：

    2025年

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  • 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

摘要

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 insight 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 insight 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.