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Direct Mapping of Polyclonal Epitopes in Serum by HDX-MS
Clint Vorauer1, Camila Boniche-Alfaro2,3, Taylor Murphree1
1Department of Medicinal Chemistry, University of Washington, Seattle, Washington 98195, United States.
Analytical Chemistry
|October 22, 2024
Summary
Researchers developed a new method using immobilized antigen and hydrogen/deuterium exchange with mass spectrometry (HDX-MS) to map antibody binding sites on antigens. This technique reveals detailed epitope landscapes in complex serum environments.
Area of Science:
- Immunology
- Structural Biology
- Analytical Chemistry
Background:
- Understanding antibody-antigen interactions is crucial for antibody-mediated protection and immunogen design.
- Existing structural techniques primarily focus on monoclonal antibodies, limiting the study of polyclonal antibody responses.
- Mapping the full epitope scope targeted by polyclonal sera in natural responses remains challenging.
Purpose of the Study:
- To develop a novel approach for mapping epitope targeting by polyclonal sera.
- To probe antibody recognition in a native serum environment.
- To analyze epitope landscapes of complex antibody responses.
Main Methods:
- Development of an immobilized antigen platform.
- Coupling immobilized antigen with hydrogen/deuterium exchange with mass spectrometry (HDX-MS).
- Application to the well-characterized model system, *Staphylococcal* enterotoxin B (SEB).
Main Results:
- Successfully detected complex combinations of epitopes targeted by polyclonal sera.
- Discerned subtle differences in epitope targeting across various antisera.
- Provided new insights into how neutralizing antibodies and antisera target SEB.
Conclusions:
- Established a novel method for directly mapping the epitope landscape of polyclonal sera.
- The developed HDX-MS approach enables analysis in a native serum environment.
- This technique offers a powerful tool for understanding natural antibody responses and designing immunogens.
