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

STAR Protoc. 2022 Feb 22;3(1):101229. doi: 10.1016/j.xpro.2022.101229. eCollection 2022 Mar 18.

Abstract

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).

Keywords: Antibody; Cell Biology; Cell-based Assays; Flow Cytometry/Mass Cytometry; Health Sciences; Microbiology; Molecular Biology.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibody Formation
  • COVID-19*
  • Flow Cytometry / methods
  • HEK293 Cells
  • Humans
  • Membrane Proteins
  • SARS-CoV-2*
  • Spike Glycoprotein, Coronavirus

Substances

  • Membrane Proteins
  • Spike Glycoprotein, Coronavirus
  • spike protein, SARS-CoV-2

Supplementary concepts

  • SARS-CoV-2 variants