Development of an In-House SARS-CoV-2 Pseudovirus-Based Neutralization Assay: A Proof of Concept Study
Development of In-House SARS-CoV-2 Pseudovirus-Based Neutralization Assay
Keywords:
SARS-CoV-2, COVID-19, Neutralizing antibodies, Pseudovirus, pVNTAbstract
The pseudovirus-based neutralization test (pVNT) is a valuable tool for detecting neutralizing antibodies against specific viruses, offering a safer and more versatile alternative to live virus assays. This project aimed to develop an in-house pVNT to detect neutralizing antibodies against SARS-CoV-2. A pseudovirus was engineered using a lentiviral system designed to express the wild-type SARS-CoV-2 spike protein and a GFP reporter gene. Key parameters for the assay, including cell number, pseudovirus dose, and detection time, were optimized. Performing this pVNT assay on SARS-CoV-2 convalescent and COVID-19 vaccinated sera comparison to plaque reduction neutralization test (PRNT) revealed a notably high correlation coefficient of 0.91. Moreover, the WHO International Standard for anti-SARS-CoV-2 immunoglobulin was tested in the pVNT to facilitate comparison of results in international units per milliliter (IU/mL). Our in-house pVNT demonstrates significant potential for detecting SARS-CoV-2 neutralizing antibodies and offers a promising alternative method for COVID-19 research. However, further studies are required to modify the pseudovirus spike protein to address ongoing variants of concern.
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