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. 2025:2937:33-59.
doi: 10.1007/978-1-0716-4591-8_3.

Mapping Antibody Epitopes by Solution NMR Spectroscopy: Practical Considerations

Elia Tamagnini  1 , Luca Simonelli  1 , Mattia Pedotti  1 , Luca Varani  2
Affiliations

Mapping Antibody Epitopes by Solution NMR Spectroscopy: Practical Considerations

Elia Tamagnini et al. Methods Mol Biol. 2025.

Abstract

Identifying an epitope, the region of the antigen in contact with an antibody, is useful in both basic and pharmaceutical research, as well as in vaccine design. Solution NMR spectroscopy is particularly well suited to residue-level characterization of intermolecular interfaces, including antibody-antigen interactions, and thus to epitope identification. Here we describe the use of NMR for the characterization of protein epitopes, focusing on experimental protocols and practical considerations, highlighting the advantages and drawbacks of the approach.

Keywords: Allosteric effects; Antibody; Antigen; Chemical shift mapping; Chemical shift perturbation; Epitope mapping; Solution NMR.

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