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Related Experiment Video

Updated: Jun 15, 2025

Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection
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Development and Validation of Novel Cell-free Direct Neutralization Assay for SARS-CoV-2.

Ji Youn Lim1, Alyssa Fiore2, Bruce Le1

  • 1AMDI Menlo Park, 3475 Edison Way, Menlo Park, CA 94025, USA.

Journal of Virological Methods
|June 11, 2025
PubMed
Summary

A new Q-NAb IgG Test quantifies neutralizing antibodies against SARS-CoV-2 variants, offering a reliable alternative to live-virus assays. This test shows strong correlation with established methods and is suitable for BSL-2 labs.

Keywords:
SARS-CoV-2 Neutralizing AntibodiesSerological assayStandardizationValidated neutralization assayWHO International Standards

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Area of Science:

  • Immunology
  • Virology
  • Assay Development

Background:

  • Neutralizing antibody titers are key indicators of protection against SARS-CoV-2.
  • Live-virus neutralization assays, the gold standard, have limitations including BSL-3 requirements and adaptability issues with new variants.

Purpose of the Study:

  • To develop and validate a novel assay, the Q-NAb IgG Test, for quantitative determination of neutralizing antibodies against SARS-CoV-2 variants.
  • To establish an assay traceable to WHO International Standards that overcomes the limitations of live-virus assays.

Main Methods:

  • Developed a Q-NAb IgG Test using a novel Fusion Protein to mimic Spike-ACE2 interaction and block non-neutralizing antibodies.
  • Measured direct binding of residual neutralizing antibodies to variant Receptor Binding Domains (RBDs) using a fluorescent secondary antibody.
  • Validated analytical performance (sensitivity, linearity, precision, interference, specificity) and clinical performance using various serum and plasma samples.

Main Results:

  • The Q-NAb IgG Test demonstrated strong agreement with live-virus microneutralization assays for both Ancestral and Omicron BA.5 strains (Spearman's correlation ρ = 0.87 and 0.92).
  • Established analytical performance characteristics and clinical sensitivity/specificity.
  • The assay is traceable to WHO International Standards.

Conclusions:

  • The Q-NAb IgG Test is a reliable, quantitative method for measuring neutralizing antibodies against SARS-CoV-2 variants.
  • Its advantages include strong correlation with live-virus assays, traceability, a convenient microtiter plate format, low sample volume, and BSL-2 compatibility.