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Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope

Hannah L Turner1, Abigail M Jackson1, Sara T Richey1

  • 1Department of Integrative Structural and Computational Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.

STAR Protocols
|July 30, 2023
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Summary

Electron microscopy-based polyclonal epitope mapping (EMPEM) rapidly assesses antibody responses to glycoprotein antigens after vaccination or infection. This method enables high-throughput analysis of antibody specificities for improved vaccine and infection studies.

Keywords:
AntibodyImmunologyMicroscopyStructural Biology

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Area of Science:

  • Immunology
  • Microscopy
  • Biochemistry

Background:

  • Antibody epitope specificity is crucial for understanding immune responses to vaccination and infection.
  • Current methods for epitope mapping can be time-consuming and lack high-throughput capabilities.
  • Glycoprotein antigens are common targets in infectious diseases and vaccine development.

Purpose of the Study:

  • To present a protocol for rapid, high-throughput electron microscopy-based polyclonal epitope mapping (EMPEM).
  • To enable detailed assessment of antibody responses to glycoprotein antigens in vaccination and infection studies.

Main Methods:

  • Detailed protocol for antibody isolation, digestion, and antigen complex purification.
  • Procedures for electron microscope imaging and EMPEM data processing and analysis.
  • Adaptation of EMPEM for high-throughput assessment of antibody responses.

Main Results:

  • Demonstration of EMPEM as a rapid and high-throughput method for epitope mapping.
  • Successful delineation of antibody specificities against glycoprotein antigens.
  • Application of the protocol for vaccination and infection studies.

Conclusions:

  • EMPEM provides a powerful tool for rapid, high-throughput epitope mapping of antibody responses.
  • This protocol facilitates detailed analysis of antibody specificities to glycoprotein antigens.
  • EMPEM is valuable for advancing vaccine and infection research.