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Immunocomplexed Antigen Capture and Identification by Native Top-Down Mass Spectrometry.
- Source :
-
Journal of the American Society for Mass Spectrometry [J Am Soc Mass Spectrom] 2023 Oct 04; Vol. 34 (10), pp. 2093-2097. Date of Electronic Publication: 2023 Sep 08. - Publication Year :
- 2023
-
Abstract
- Antibody-antigen interactions are central to the immune response. Variation of protein antigens such as isoforms and post-translational modifications can alter their antibody binding sites. To directly connect the recognition of protein antigens with their molecular composition, we probed antibody-antigen complexes by using native tandem mass spectrometry. Specifically, we characterized the prominent peanut allergen Ara h 2 and a convergent IgE variable region discovered in patients who are allergic to peanuts. In addition to measuring the antigen-induced dimerization of IgE antibodies, we demonstrated how immunocomplexes can be isolated in the gas phase and activated to eject, identify, and characterize proteoforms of their bound antigens. Using tandem experiments, we isolated the ejected antigens and then fragmented them to identify their chemical composition. These results establish native top-down mass spectrometry as a viable platform for precise and thorough characterization of immunocomplexes to relate structure to function and enable the discovery of antigen proteoforms and their binding sites.
Details
- Language :
- English
- ISSN :
- 1879-1123
- Volume :
- 34
- Issue :
- 10
- Database :
- MEDLINE
- Journal :
- Journal of the American Society for Mass Spectrometry
- Publication Type :
- Academic Journal
- Accession number :
- 37683262
- Full Text :
- https://doi.org/10.1021/jasms.3c00235