Simultaneous Measurement of IgM and IgG Antibodies to SARSCoV- 2 Spike, RBD, and Nucleocapsid Multiplexed in a Single Assay on the xMAP INTELLIFLEX DR-SE Flow Analyzer

Andrew Cameron, Jessica L. Bohrhunter, Claire A. Porterfield, Rupinder Mangat, Michael H. Karasick, Zachary Pearson, Stephen Angeloni, Nicole D. Pecora

Research output: Contribution to journalArticlepeer-review

Abstract

The multiplex capabilities of the new xMAP INTELLIFLEX DR-SE flow analyzer were explored by modifying a serological assay previously used to characterize the IgG antibody to infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The goal was to examine the instrument's performance and to simultaneously measure IgM and IgG antibody responses against multiple SARS-CoV-2 antigens in a single assay. Specific antibodies against the SARS-CoV-2 spike (S), receptor binding domain (RBD), and nucleocapsid (N) proteins were investigated in 310 symptomatic case patients using a fluorescent microsphere immunoassay and simultaneous detection of IgM and IgG. Neutralization potential was studied using the addition of soluble angiotensin- converting enzyme 2 (ACE2) to block antibody binding. A profile extending to 180 days from symptom onset (DFSO) was described for antibodies specific to each viral antigen. Generally, IgM levels peaked and declined rapidly ;3-4 weeks following infection, whereas S- and RBD-specific IgG plateaued at 80 DFSO. ACE2 more effectively prevented IgM and IgG binding in convalescent cases . 30 DFSO, suggesting those antibodies had greater neutralization potential. This work highlighted the multiplex and multi-analyte potential of the xMAP INTELLIFLEX DR-SE, and provided further evidence for antigen-specific IgM and IgG trajectories in acute and convalescent cases.

Original languageEnglish (US)
JournalMicrobiology Spectrum
Volume10
Issue number2
DOIs
StatePublished - Apr 2022
Externally publishedYes

Keywords

  • ACE2
  • antibody
  • COVID-19
  • fluorescent microsphere immunoassay
  • FMIA
  • microsphere
  • nucleocapsid
  • SARS-CoV-2
  • serology
  • spike glycoprotein

ASJC Scopus subject areas

  • Physiology
  • Ecology
  • Immunology and Microbiology(all)
  • Genetics
  • Microbiology (medical)
  • Cell Biology
  • Infectious Diseases

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