Quantitative and Standardized Pseudovirus Neutralization Assay for COVID-19

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2779; p. 259
• Main Authors: Izac, Jerilyn R, Kwee, Edward J, Gaigalas, Adolfas, Wang, Lili
• Format: Journal Article
• Language: English
• Published: United States 2024
• Subjects: Antibodies, Neutralizing; Antibodies, Viral; COVID-19; COVID-19 Serotherapy; Flow Cytometry; Humans; Immunologic Tests; Neutralization Tests; SARS-CoV-2; Spike Glycoprotein, Coronavirus; Flow cytometry; SARS-CoV-2; WHO International Standard; HEK293-hACE2-TMPRSS2-mCherry cell line; Quantification; Neutralizing antibodies; Live cell imaging; Multiplicity of infection (MOI); Pseudovirus neutralization assay; Assay standardization; VSV-ΔG spike pseudovirus; NIBSC 20/136 (WHO IS 20/136)
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-3738-8_11

COVID-19 is a global pandemic caused by the highly infectious SARS-CoV-2 virus. Efforts to combat SARS-CoV-2 infection include mass vaccination and development of monoclonal and convalescent plasma therapeutics that require precise measurements of correlative, functional neutralizing antibodies that prevent virus infection. Developing rapid, safe, easy-to-use, and high-quality neutralization assays are essential for the success of the massive effort. Here, we developed a vesicular stomatitis virus-based neutralization assay that was capable of quantifying varying degrees of neutralization in patient serum samples. This assay has two detection readouts, flow cytometry and live cell imaging. The two readout methods produced consistent values of all 50% neutralization titers, further enhancing measurement confidence on the assay. Moreover, the use of available reference standards such as the World Health Organization International Standard (NIBSC code 20/136) enables quantification and standardization of the pseudovirus neutralization assay with neutralizing antibody titers measured in International Units/mL. Quantitative and standardized neutralization assays are critical for reliable efficacy evaluation and comparison of numerous vaccines and therapeutics.