Quantitative and Standardized Pseudovirus Neutralization Assay for COVID-19.
Methods Mol Biol
; 2779: 259-271, 2024.
Article
en En
| MEDLINE
| ID: mdl-38526789
ABSTRACT
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.
Palabras clave
Assay standardization; Flow cytometry; HEK293-hACE2-TMPRSS2-mCherry cell line; Live cell imaging; Multiplicity of infection (MOI); NIBSC 20/136 (WHO IS 20/136); Neutralizing antibodies; Pseudovirus neutralization assay; Quantification; SARS-CoV-2; VSV-ΔG spike pseudovirus; WHO International Standard
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
COVID-19
Límite:
Humans
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2024
Tipo del documento:
Article
País de afiliación:
Estados Unidos