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Related Experiment Video

Updated: Aug 15, 2025

Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection

Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection

Published on: June 5, 2021

A high-throughput SARS-CoV-2 pseudovirus multiplex neutralization assay.

Benjamin Louis Sievers¹, Terri Gelbart¹, Gene S Tan²

¹J. Craig Venter Institute, La Jolla, CA 92037, USA.

|January 3, 2023

Summary

This summary is machine-generated.

This study presents a novel assay for measuring neutralizing antibody titers against SARS-CoV-2 variants of concern (VOCs). The method uses dual-fluorophore pseudoviruses, enabling efficient and reliable assessment of vaccine-induced protection.

Keywords:

Antibody Cell-based Assays High Throughput Screening Microbiology

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Area of Science:

Virology
Immunology
Vaccinology

Background:

Accurate measurement of neutralizing antibody titers against SARS-CoV-2 is crucial for understanding vaccine effectiveness.
Existing methods can be time-consuming and resource-intensive, necessitating more efficient approaches.

Purpose of the Study:

To develop and validate a novel assay for quantifying neutralizing antibody titers against SARS-CoV-2 variants of concern (VOCs).
To enable simultaneous assessment of neutralization against multiple VOCs in a single assay run.

Main Methods:

Utilized single-cycle vesicular stomatitis virus (VSV) pseudoviruses engineered to display spike (S) proteins from SARS-CoV-2 VOCs.
Incorporated two distinct fluorophores, each linked to the S protein of a different VOC, allowing for multiplexed detection.
Determined neutralization titers by measuring the reduction in fluorescent signal indicative of viral entry inhibition.

Main Results:

The developed assay allows for the simultaneous determination of neutralization titers against two different VOCs in a single experimental run.
This approach offers a generalizable method that significantly reduces assay time, sample consumption, and run-to-run variability.

Conclusions:

This novel dual-fluorophore pseudovirus assay provides an efficient and reliable platform for evaluating neutralizing antibody responses to SARS-CoV-2 VOCs.
The assay's generalizability and efficiency make it a valuable tool for vaccine development and monitoring population immunity.