Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2025:2937:61-77.
doi: 10.1007/978-1-0716-4591-8_4.

Cryo-Electron Microscopy-Based Epitope Mapping

Ieva Drulyte  1   2 , Daniel L Hurdiss  3
Affiliations

Cryo-Electron Microscopy-Based Epitope Mapping

Ieva Drulyte et al. Methods Mol Biol. 2025.

Abstract

Antibodies play a pivotal role in our adaptive immune system and serve as a popular format for drug development. Understanding the molecular mechanisms underlying antibodies binding to their specific antigens is relevant for therapeutic and vaccine development, as well as for gaining insights into the recognition of self and non-self-antigens. Cryo-electron microscopy (cryo-EM) single-particle analysis (SPA) has emerged as a powerful technique to study the interactions between antibodies and a diverse array of biological targets. In this chapter, we provide a generic protocol, accompanied by valuable tips and tricks, specifically tailored for the examination of antigen-antibody complexes using the cryo-EM SPA methodology.

Keywords: Antibodies; Cryo-electron microscopy; Epitope mapping; Epitope-Paratope; Protein-protein interactions; Single-particle analysis; Structural biology.

PubMed Disclaimer

References

    1. Renaud JP, Chari A, Ciferri C et al (2018) Cryo-EM in drug discovery: achievements, limitations and prospects. Nat Rev Drug Discov 177(17):471–492. https://doi.org/10.1038/nrd.2018.77 - DOI
    1. Nakane T, Kotecha A, Sente A et al (2020) Single-particle cryo-EM at atomic resolution. Nature 587:152–156. https://doi.org/10.1038/s41586-020-2829-0 - DOI - PubMed - PMC
    1. Weissenberger G, Henderikx RJM, Peters PJ (2021) Understanding the invisible hands of sample preparation for cryo-EM. Nat Methods 18:463–471. https://doi.org/10.1038/S41592-021-01130-6 - DOI - PubMed
    1. Zivanov J, Nakane T, Forsberg BO et al (2018) New tools for automated high-resolution cryo-EM structure determination in RELION-3. eLife 7:e42166. https://doi.org/10.7554/eLife.42166
    1. Punjani A, Rubinstein JL, Fleet DJ, Brubaker MA (2017) CryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat Methods 14:290–296. https://doi.org/10.1038/nmeth.4169

MeSH terms

LinkOut - more resources