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

• Published in: STAR protocols Vol. 3; no. 1; p. 101229
• Main Authors: Martin, Sophie, Jégou, Gwénaële, Nicolas, Aurore, Le Gallo, Matthieu, Chevet, Éric, Godey, Florence, Avril, Tony
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 18.03.2022; Elsevier
• 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; Microbiology; Molecular Biology; Antibody; Flow Cytometry/Mass Cytometry; Cell-based Assays; Cell Biology; Health Sciences
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2022.101229

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.