A cell-based system combined with flow cytometry to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in patients with COVID-19

This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing H...

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Bibliographic Details
Published in:STAR protocols 2022-03, Vol.3 (1), p.101229-101229, Article 101229
Main Authors: Martin, Sophie, Jégou, Gwénaële, Nicolas, Aurore, Le Gallo, Matthieu, Chevet, Éric, Godey, Florence, Avril, Tony
Format: Article
Language:English
Subjects:
Antibody
Antibody Formation
Cancer
Cell Biology
Cell-based Assays
COVID-19
Flow Cytometry - methods
Flow Cytometry/Mass Cytometry
Health Sciences
HEK293 Cells
Humans
Life Sciences
Membrane Proteins
Microbiology
Molecular Biology
Protocol
SARS-CoV-2
Spike Glycoprotein, Coronavirus
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Summary:This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness. For complete details on the use and execution of this protocol, please refer to Martin et al. (2021). [Display omitted] •A human-cell-based assay was developed to access antibody responses against SARS-CoV-2•The assay recapitulates spike membrane insertion and posttranslational modifications•The assay represents a powerful tool to test spike variants and vaccine effectiveness This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness.
ISSN:2666-1667
2666-1667
DOI:10.1016/j.xpro.2022.101229