Measuring Neutralizing Antibodies to SARS-CoV-2 Using Lentiviral Spike-Pseudoviruses

Sabari Nath Neerukonda; Russell Vassell; Carol D Weiss; Wei Wang

doi:10.1007/978-1-0716-2111-0_18

Measuring Neutralizing Antibodies to SARS-CoV-2 Using Lentiviral Spike-Pseudoviruses

Methods Mol Biol. 2022:2452:305-314. doi: 10.1007/978-1-0716-2111-0_18.

Authors

Sabari Nath Neerukonda¹, Russell Vassell¹, Carol D Weiss², Wei Wang³

Affiliations

¹ Office of Vaccines Research and Review, Center for Biologics Evaluation and Research and Review, US Food and Drug Administration, Silver Spring, MD, USA.
² Office of Vaccines Research and Review, Center for Biologics Evaluation and Research and Review, US Food and Drug Administration, Silver Spring, MD, USA. carol.weiss@fda.hhs.gov.
³ Office of Vaccines Research and Review, Center for Biologics Evaluation and Research and Review, US Food and Drug Administration, Silver Spring, MD, USA. wei.wang@fda.hhs.gov.

PMID: 35554914
DOI: 10.1007/978-1-0716-2111-0_18

Abstract

Assays measuring neutralizing antibodies (nAbs) against SARS-CoV-2 are used to evaluate serological responses after SARS-CoV-2 infection and the potency of therapeutic antibodies and preventive vaccines. It is therefore imperative that neutralization assays be sensitive, specific, quantitative, and scalable for high throughput. Pseudoviruses are excellent surrogates for highly pathogenic viruses such as SARS-CoV-2 because they can be safely used to measure nAbs in a biosafety level-2 laboratory. In addition, pseudoviruses allow for easy introduction of mutations to study the effect of amino acid changes in the spike protein. In this chapter, we describe a recently optimized assay for measuring neutralizing antibodies to SARS-CoV-2 that uses a HIV-based lentiviral vector pseudotyped with the spike glycoprotein of SARS-CoV-2 to infect 293T cells stably expressing ACE2 and TMPRSS2.

Keywords: Assay optimization; Luciferase reporter; Neutralizing antibodies; Pseudovirus; SARS-CoV-2.

MeSH terms

Antibodies, Neutralizing
Antibodies, Viral
COVID-19*
Humans
Neutralization Tests
SARS-CoV-2* / genetics
Spike Glycoprotein, Coronavirus / genetics

Substances

Antibodies, Neutralizing
Antibodies, Viral
Spike Glycoprotein, Coronavirus
spike protein, SARS-CoV-2