Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping

Electron microscopy-based polyclonal epitope mapping (EMPEM) can delineate epitope specificities of serum antibodies to a given antigen following vaccination or infection. Here, we present a protocol for the EMPEM method for rapid high-throughput assessment of antibody responses to glycoprotein anti...

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Autores principales: Turner, Hannah L., Jackson, Abigail M., Richey, Sara T., Sewall, Leigh M., Antanasijevic, Aleksandar, Hangartner, Lars, Ward, Andrew B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10400963/
https://www.ncbi.nlm.nih.gov/pubmed/37516970
http://dx.doi.org/10.1016/j.xpro.2023.102476
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author Turner, Hannah L.
Jackson, Abigail M.
Richey, Sara T.
Sewall, Leigh M.
Antanasijevic, Aleksandar
Hangartner, Lars
Ward, Andrew B.
author_facet Turner, Hannah L.
Jackson, Abigail M.
Richey, Sara T.
Sewall, Leigh M.
Antanasijevic, Aleksandar
Hangartner, Lars
Ward, Andrew B.
author_sort Turner, Hannah L.
collection PubMed
description Electron microscopy-based polyclonal epitope mapping (EMPEM) can delineate epitope specificities of serum antibodies to a given antigen following vaccination or infection. Here, we present a protocol for the EMPEM method for rapid high-throughput assessment of antibody responses to glycoprotein antigens in vaccination and infection studies. We describe steps for antibody isolation and digestion, antigen complex and purification, and electron microscope imaging. We then detail procedures for processing and analysis of EMPEM data. For complete details on the use and execution of this protocol, please refer to Bianchi et al. (2018).(1)
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spelling pubmed-104009632023-08-05 Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping Turner, Hannah L. Jackson, Abigail M. Richey, Sara T. Sewall, Leigh M. Antanasijevic, Aleksandar Hangartner, Lars Ward, Andrew B. STAR Protoc Protocol Electron microscopy-based polyclonal epitope mapping (EMPEM) can delineate epitope specificities of serum antibodies to a given antigen following vaccination or infection. Here, we present a protocol for the EMPEM method for rapid high-throughput assessment of antibody responses to glycoprotein antigens in vaccination and infection studies. We describe steps for antibody isolation and digestion, antigen complex and purification, and electron microscope imaging. We then detail procedures for processing and analysis of EMPEM data. For complete details on the use and execution of this protocol, please refer to Bianchi et al. (2018).(1) Elsevier 2023-07-28 /pmc/articles/PMC10400963/ /pubmed/37516970 http://dx.doi.org/10.1016/j.xpro.2023.102476 Text en © 2023 The Author(s) https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Turner, Hannah L.
Jackson, Abigail M.
Richey, Sara T.
Sewall, Leigh M.
Antanasijevic, Aleksandar
Hangartner, Lars
Ward, Andrew B.
Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping
title Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping
title_full Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping
title_fullStr Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping
title_full_unstemmed Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping
title_short Protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping
title_sort protocol for analyzing antibody responses to glycoprotein antigens using electron-microscopy-based polyclonal epitope mapping
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10400963/
https://www.ncbi.nlm.nih.gov/pubmed/37516970
http://dx.doi.org/10.1016/j.xpro.2023.102476
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