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

STAR Protoc. 2023 Sep 15;4(3):102476. doi: 10.1016/j.xpro.2023.102476. Epub 2023 Jul 28.

Abstract

Electron microscopy-based polyclonal epitope mapping (EMPEM) can delineate epitope specificities of serum antibodies to a given antigen following vaccination or infection. Here, we present a protocol for the EMPEM method for rapid high-throughput assessment of antibody responses to glycoprotein antigens in vaccination and infection studies. We describe steps for antibody isolation and digestion, antigen complex and purification, and electron microscope imaging. We then detail procedures for processing and analysis of EMPEM data. For complete details on the use and execution of this protocol, please refer to Bianchi et al. (2018).1.

Keywords: Antibody; Immunology; Microscopy; Structural Biology.

MeSH terms

  • Antibodies
  • Antibody Formation*
  • Electrons*
  • Epitope Mapping
  • Glycoproteins
  • Microscopy, Electron

Substances

  • Antibodies
  • Glycoproteins