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

使用基于电子显微镜的多克隆表位图谱分析对糖蛋白抗原的抗体反应的方案

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作者:Hannah L Turner, Abigail M Jackson, Sara T Richey, Leigh M Sewall, Aleksandar Antanasijevic, Lars Hangartner, Andrew B Ward

Abstract

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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