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Updated: Jul 23, 2026
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Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection
Published on: June 5, 2021
4.6K
Development of a SARS-CoV-2 neutralization assay based on a pseudotyped virus using a HIV system
Ziteng Liang1,2, Jincheng Tong2, Xi Wu2
1Chinese Academy of Medical Sciences & Peking Union Medical College Dongcheng District, Beijing China.
Medcomm
|March 25, 2024
Summary
A new fluorescent pseudovirus neutralization assay for SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) was developed. This assay is accurate, sensitive, and scalable for detecting neutralizing antibodies, aiding vaccine development.
Area of Science:
- Virology
- Immunology
- Biotechnology
Background:
- Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) poses a global health threat.
- Pseudovirus neutralization assays are valuable tools for studying SARS-CoV-2 due to their safety and efficiency in BSL-2 laboratories.
Purpose of the Study:
- To develop a robust and optimized fluorescent pseudotyped virus neutralization assay for SARS-CoV-2.
- To establish a reliable method for detecting neutralizing antibodies against SARS-CoV-2.
Main Methods:
- Engineered a high-titer fluorescent pseudovirus (SARS-CoV-2-Sdel21-AmCyan) using an HIV backbone and a modified SARS-CoV-2 spike protein.
- Established and optimized a neutralization assay using 293T AF cells in 96-well plates.
- Validated the assay's performance and scaled it up to 384-well plates.
Main Results:
- Successfully packaged high-titer fluorescent pseudovirus.
- Optimized key assay parameters including cell density, virus inoculation, and incubation times.
- Demonstrated high accuracy, sensitivity, repeatability, and correlation with luminescent assays.
Conclusions:
- A robust fluorescent pseudotyped virus neutralization assay for SARS-CoV-2 has been established.
- This assay provides a reliable platform for serum neutralization antibody detection, monoclonal antibody screening, and vaccine development.
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