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Related Experiment Video

Updated: Nov 18, 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

Detecting SARS-CoV-2 3CLpro expression and activity using a polyclonal antiserum and a luciferase-based biosensor.

Amornrat O'Brien¹, Da-Yuan Chen², Matthew Hackbart¹

¹Department of Microbiology and Immunology, Loyola University Chicago, Stritch School of Medicine, Maywood, IL, USA.

|February 6, 2021

Summary

Researchers developed a new assay to screen for SARS-CoV-2 inhibitors targeting the 3CLpro protease, crucial for blocking viral replication and aiding COVID-19 drug development.

Keywords:

3CLpro Main protease Mpro Protease inhibitors SARS-CoV-2 nsp5 pGlo biosensor

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Last Updated: Nov 18, 2025

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Area of Science:

Virology
Drug Discovery
Biochemistry

Background:

The COVID-19 pandemic necessitates the development of antivirals targeting SARS-CoV-2.
The SARS-CoV-2 3C-like protease (3CLpro) is essential for viral replication and a key drug target.

Purpose of the Study:

To develop novel reagents for screening SARS-CoV-2 3CLpro inhibitors.
To facilitate the pre-clinical evaluation of potential antiviral drugs against COVID-19.

Main Methods:

Developed a luminescence-based biosensor assay to evaluate small molecule inhibitors of SARS-CoV-2 3CLpro.
Utilized a polyclonal rabbit antiserum against SARS-CoV 3CLpro, demonstrating cross-reactivity with SARS-CoV-2 3CLpro.

Main Results:

Successfully established a functional luminescence-based assay for SARS-CoV-2 3CLpro.
Confirmed cross-reactivity of existing antiserum, enabling its use for SARS-CoV-2 research.

Conclusions:

The developed biosensor assay and cross-reactive antiserum are valuable tools for identifying and evaluating SARS-CoV-2 protease inhibitors.
These reagents will accelerate the pre-clinical development of COVID-19 therapeutics.