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Mol Cell Proteomics ; 23(3): 100734, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38342408

RESUMEN

Antigen-antibody interactions play a key role in the immune response post vaccination and the mechanism of action of antibody-based biopharmaceuticals. 4CMenB is a multicomponent vaccine against Neisseria meningitidis serogroup B in which factor H binding protein (fHbp) is one of the key antigens. In this study, we use hydrogen/deuterium exchange mass spectrometry (HDX-MS) to identify epitopes in fHbp recognized by polyclonal antibodies (pAb) from two human donors (HDs) vaccinated with 4CMenB. Our HDX-MS data reveal several epitopes recognized by the complex mixture of human pAb. Furthermore, we show that the pAb from the two HDs recognize the same epitope regions. Epitope mapping of total pAb and purified fHbp-specific pAb from the same HD reveals that the two antibody samples recognize the same main epitopes, showing that HDX-MS based epitope mapping can, in this case at least, be performed directly using total IgG pAb samples that have not undergone Ab-selective purification. Two monoclonal antibodies (mAb) were previously produced from B-cell repertoire sequences from one of the HDs and used for epitope mapping of fHbp with HDX-MS. The epitopes identified for the pAb from the same HD in this study, overlap with the epitopes recognized by the two individual mAbs. Overall, HDX-MS epitope mapping appears highly suitable for simultaneous identification of epitopes recognized by pAb from human donors and to thus both guide vaccine development and study basic human immunity to pathogens, including viruses.


Asunto(s)
Infecciones Meningocócicas , Vacunas Meningococicas , Neisseria meningitidis , Humanos , Mapeo Epitopo/métodos , Neisseria meningitidis/metabolismo , Deuterio/metabolismo , Proteínas Bacterianas/metabolismo , Infecciones Meningocócicas/prevención & control , Proteínas Portadoras , Medición de Intercambio de Deuterio , Factor H de Complemento , Antígenos Bacterianos , Epítopos , Anticuerpos Monoclonales/metabolismo , Espectrometría de Masas de Intercambio de Hidrógeno-Deuterio
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