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Protein Engineering by Yeast Surface Display
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Epitope Mapping Using Yeast Display and Next Generation Sequencing.

Thomas Van Blarcom1, Andrea Rossi2, Davide Foletti2,3

  • 1Rinat, Pfizer Inc., South San Francisco, CA, USA. thomas.vanblarcom@pfizer.com.

Methods in Molecular Biology (Clifton, N.J.)
|May 2, 2018
PubMed
Summary
This summary is machine-generated.

We developed a rapid method to precisely map antibody epitopes using yeast display and DNA sequencing. This technique provides quantitative insights into critical binding residues for therapeutic proteins like monoclonal antibodies.

Keywords:
Alpha toxinAntibodyAntigenEpitope mappingFACSLibrary designNext generation sequencingStaphylococcus aureusYeast display

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Area of Science:

  • Biochemistry and Molecular Biology
  • Immunology
  • Protein Engineering

Background:

  • Monoclonal antibodies are a major class of therapeutic proteins due to their high affinity and specificity for antigens.
  • Precise epitope determination is crucial for understanding antibody mechanisms and differentiating similar antibodies.

Purpose of the Study:

  • To describe a novel method for precise and efficient parallel epitope mapping of multiple antibodies.
  • To gain quantitative insights into critical epitope residues involved in antibody-antigen interactions.

Main Methods:

  • A combination of rational library design and yeast surface display was employed.
  • Next-generation DNA sequencing was utilized for high-throughput analysis.
  • The method was validated by mapping epitopes of antibodies neutralizing Staphylococcus aureus alpha toxin.

Main Results:

  • The described method enables precise and efficient parallel epitope mapping within weeks.
  • Quantitative insights into key epitope residues essential for antibody binding were obtained.
  • Successful application demonstrated for antibodies targeting alpha toxin.

Conclusions:

  • This approach offers a significant advancement in characterizing antibody-antigen interactions.
  • The method facilitates a deeper understanding of therapeutic antibody function and development.
  • It provides a powerful tool for antibody characterization and differentiation.