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Mapping antibody binding using multiplexed epitope substitution analysis.

Michael L Paull1, Joel D Bozekowski1, Patrick S Daugherty1

  • 1Department of Chemical Engineering, University of California, Santa Barbara, CA 93106, USA.

Journal of Immunological Methods
|November 10, 2021
PubMed
Summary

Multiplexed Epitope Substitution Analysis (MESA) rapidly maps antibody binding sites. This new method accelerates the development of diagnostics, therapeutics, and vaccines by revealing precise epitope binding motifs.

Keywords:
Antibody bindingB cell epitopesEpitope mappingNext-generation sequencingRandom peptide librariesSaturation mutagenesis

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Area of Science:

  • Immunology
  • Biotechnology
  • Computational Biology

Background:

  • Understanding antibody epitopes is crucial for developing diagnostics, therapeutics, and vaccines.
  • Current epitope mapping methods are limited by low throughput and require targeted experimental approaches.

Purpose of the Study:

  • To develop a high-throughput method for characterizing antibody epitopes.
  • To accelerate the discovery and understanding of antibody-epitope interactions.

Main Methods:

  • Developed Multiplexed Epitope Substitution Analysis (MESA) to screen millions of antibody-binding peptides.
  • Utilized next-generation sequencing (NGS) to determine peptide sequences.
  • Employed computational analysis of k-mer subsequences and amino acid substitutions to identify binding motifs.

Main Results:

  • Validated MESA by accurately determining binding motifs for monoclonal antibodies.
  • Characterized population-level epitope binding motifs from 50 serum specimens against common pathogens.
  • Revealed unique binding signatures for HSV-1 seropositive specimens, demonstrating MESA's ability to capture population variability.

Conclusions:

  • MESA enables rapid identification and characterization of binding motifs for numerous epitopes in a single experiment.
  • This method significantly accelerates discoveries in antibody-epitope interactions.
  • MESA enhances the development of targeted diagnostics, therapeutics, and vaccines.