Cryo-Electron Microscopy-Based Epitope Mapping

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.