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Related Experiment Video

Updated: Jan 17, 2026

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 Safe and Accessible Cell-Based Spike-ACE2 Binding Assay for Evaluating SARS-CoV-2 Neutralization Activity in

Martin A Rossotti¹, Shannon Ryan¹, Greg Hussack¹

¹Human Health Therapeutics Research Centre, National Research Council Canada, Ottawa, ON K1A 0R6, Canada.

Methods and Protocols

|September 22, 2025

Summary

This summary is machine-generated.

A new cell-based assay using the SARS-CoV-2 spike protein and ACE2 interaction can identify neutralizing antibodies without live virus. This method enhances laboratory biosafety and correlates well with traditional neutralization assays.

Keywords:

ACE2 SARS-CoV-2 antibodies flow cytometry nanobodies neutralization neutralizing antibodies (nAbs)serum spike vaccination

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High-throughput Confocal Imaging of Quantum Dot-Conjugated SARS-CoV-2 Spike Trimers to Track Binding and Endocytosis in HEK293T Cells

Published on: April 21, 2022

Area of Science:

Virology
Immunology
Biotechnology

Background:

Severe Acute Respiratory Syndrome-Coronavirus 2 (SARS-CoV-2) is a Risk Group 3 pathogen causing COVID-19.
Handling live SARS-CoV-2 necessitates high-level biosafety containment (CL3 facilities).
Traditional neutralization assays often require infectious viral agents, posing biosafety challenges.

Purpose of the Study:

To describe a cell-based spike-ACE2 binding assay as a safer alternative for identifying SARS-CoV-2 neutralizing antibodies.
To validate this assay as a reliable surrogate for neutralization activity.
To highlight the biosafety advantages of using a non-replicating viral agent assay.

Main Methods:

Utilized recombinant SARS-CoV-2 trimeric spike protein and human ACE2.
Quantified the binding interaction using flow cytometry.
Compared results with pseudotyped lentiviral and live virus neutralization assays.

Main Results:

The cell-based spike-ACE2 binding assay effectively identifies potential neutralizing antibodies against SARS-CoV-2.
The assay demonstrated a strong correlation with established neutralization methods.
This surrogate assay successfully identified neutralizing antibodies without using infectious viral particles.

Conclusions:

The described cell-based assay provides a biosafely enhanced method for detecting SARS-CoV-2 neutralizing antibodies.
This approach is a valuable tool for antibody screening and research, applicable beyond SARS-CoV-2.
The assay offers a reliable and safer alternative to traditional neutralization assays.