Abstract
Abstract Neutralisation assays are crucial tools to quantify the presence of functional neutralising antibodies in serum samples. Since the SARS-CoV-2 virus (the causative agent of COVID-19) is designated as a category 3 biosafety level pathogen, pseudotyped viruses bearing the SARS-CoV-2 spike protein permit extensive and widespread serum/plasma screening in a BSL 2 laboratory. These assays can be used to assess viral tropism, vaccine immunogenicity, efficacy of antiviral compounds (incl. therapeutic mAbs) and serosurveillance studies. In this article, we highlight approaches to SARS-CoV-2 viral pseudotyping, its practicality, and utility in increasing our understanding of neutralising antibodies against SARS-CoV-2.
Highlights
The Role of Pseudotype Neutralisation Assays in Understanding SARS CoV-2 Diego Cantoni1, Martin Mayora-Neto1, Nigel Temperton1, * 1 Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Kent and Greenwich at Medway, Chatham ME7 4TB, UK *Author to whom correspondence should be addressed Neutralisation assays are crucial tools to quantify the presence of functional neutralising antibodies in serum samples
Several types of viral cores are used for generating Pseudotyped viruses (PV) such as murine leukaemia virus (MLV), vesicular stomatitis virus (VSV) and, human immunodeficiency virus (HIV), all of which have been described previously 1–5
Whilst it is true that ELISA assays can be used to detect presence of antibodies in response to viral infection, the system itself detects antibody binding to the viral protein, but it does not measure whether this interaction results in neutralisation, though recently next-generation kits have been developed to provide that level of information 13–15
Summary
The Role of Pseudotype Neutralisation Assays in Understanding SARS CoV-2 Diego Cantoni[1], Martin Mayora-Neto[1], Nigel Temperton1, * 1 Viral Pseudotype Unit, Medway School of Pharmacy, The Universities of Kent and Greenwich at Medway, Chatham ME7 4TB, UK *Author to whom correspondence should be addressed Neutralisation assays are crucial tools to quantify the presence of functional neutralising antibodies in serum samples. The PV neutralization assay is typically carried out in 96-well plates, whereby serum samples are serially diluted and incubated with a fixed amount of SARS- CoV-2 PVs to allow any neutralising antibodies to bind to the Spike (S).
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