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Related Concept Videos

Enzyme-Linked Immunosorbent Assay01:33

Enzyme-Linked Immunosorbent Assay

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In 1971, Peter Perlman and Eva Engvall developed an Enzyme-linked immunosorbent assay (ELISA or EIA). ELISA differs from western blot in that the assays are conducted in microtiter plates or in vivo rather than on an absorbent membrane.
There are many different types of ELISAs, but they all involve an antibody molecule whose constant region binds an enzyme, leaving the variable region free to bind its specific antigen.  Enzyme-substrate reaction allows the antigen to be visualized or...
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Related Experiment Video

Updated: Jan 18, 2026

Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection
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A High-Throughput Broad Neutralizing Antibody Assay for Detecting SARS-CoV-2 Variant Immunity in Population.

Xiaohan Zhang1,2,3, Yajie Wang4, Mansheng Li2

  • 1Nosocomial Infection Management Department, Tianjin Medical University Cancer Institute & Hospital, National Clinical Research Center for Cancer, Tianjin's Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin 300202, China.

ACS Infectious Diseases
|September 12, 2025
PubMed
Summary

A new high-throughput assay detects neutralizing antibodies against SARS-CoV-2 variants. This tool is crucial for understanding immune escape and developing better COVID-19 vaccines and strategies.

Keywords:
COVID-19SARS-CoV-2flow cytometryneutralizing antibodyvariant

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Area of Science:

  • Immunology
  • Virology
  • Proteomics
  • Vaccinology

Background:

  • Detecting neutralizing antibodies (NAbs) against evolving SARS-CoV-2 variants is critical for assessing host immune escape.
  • Existing methods for NAb detection can be time-consuming or lack the sensitivity needed for diverse variants.

Purpose of the Study:

  • To develop a rapid, sensitive, and high-throughput assay for detecting broad neutralizing antibodies (bNAbs) against various SARS-CoV-2 variants.
  • To evaluate the assay's performance against established neutralization assays and apply it to assess NAb responses in vaccinated individuals and HIV patients.

Main Methods:

  • Development of a flow cytometry-based assay utilizing magnetic-fluorescent microspheres for detecting NAbs against diverse SARS-CoV-2 variants.
  • Validation of the assay by correlating results with IgG serological assays, cPass surrogate virus neutralization test (sVNT), pseudovirus neutralization assays, and live virus neutralization assays.
  • Application of the assay to analyze NAb responses in healthy individuals post-third-dose vaccination and in individuals with HIV experiencing breakthrough SARS-CoV-2 infections.

Main Results:

  • The developed assay demonstrated high sensitivity (35-fold greater than Luminex) and strong correlations with multiple established neutralization assays (R values ranging from 0.64 to 0.96).
  • Omicron BA.1-BA.5 variants showed significant resistance to inactivated vaccines in healthy individuals.
  • In HIV patients, breakthrough infections with Omicron variants induced broad neutralizing activity, but NAb levels were lower in individuals with decreased immunity (CD4+ < 350 cells/μL) compared to those with recovered immunity (CD4+ > 500 cells/μL).

Conclusions:

  • The high-throughput bNAb assay is a reliable and sensitive tool for detecting neutralizing antibodies against SARS-CoV-2 variants.
  • The findings highlight the immune escape of Omicron variants from inactivated vaccines and underscore the importance of maintaining immune function for effective NAb production, particularly in immunocompromised individuals.
  • This platform can inform the development of next-generation COVID-19 vaccines and vaccination strategies.