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Pseudovirus-Based Assays for the Measurement of Antibody-Mediated Neutralization of SARS-CoV-2

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SARS-CoV-2

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2452))

Abstract

SARS-CoV-2 has emerged as a significant cause of morbidity and mortality worldwide. Virus neutralization assays are critical for the development and evaluation of vaccines and immunotherapeutics, as well as for conducting basic research into the immune response, spread, and pathogenesis of this disease. However, neutralization assays traditionally require the use of infectious virus which must be carefully handled in a BSL-3 setting, thus complicating the assay and restricting its use to labs with access to BSL-3 facilities. Pseudovirus-based assays are an alternative to the use of infectious virus. SARS-CoV-2 pseudovirus contains only the spike structural protein, and infection results in a single round of replication, thus allowing for the assay to be run safely under BSL-2 conditions. In this chapter, we describe protocols and considerations for the production and titration of lentivirus-based SARS-CoV-2 pseudovirus, as well as for running and analysis of FACS-based pseudovirus neutralization assays.

Copyright Statement:

Peifang Sun, Maya Williams, Gabriel Defang, and Kevin R. Porter authors are military service members or federal/contracted employees of the U.S. Government. This work was prepared as part of their official duties. Title 17 U.S.C. 105 provides that “copyright protection under this title is not available for any work of the United States Government.” Title 17 U.S.C. 101 defines a U.S. Government work as a work prepared by a military service member or employee of the U.S. Government as part of that person’s official duties.

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Authors and Affiliations

  1. Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, USA

    Corey Balinsky & Vihasi Jani

  2. Viral and Rickettsial Diseases Department, Naval Medical Research Center, Silver Spring, MD, USA

    Peifang Sun, Gabriel Defang & Kevin R. Porter

  3. Chemistry Division, US Naval Research Laboratory, Washington, DC, USA

    Maya Williams

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  1. Corey Balinsky
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  3. Peifang Sun
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  4. Maya Williams
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  5. Gabriel Defang
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  6. Kevin R. Porter
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Editors and Affiliations

  1. Department of Microbiology and Immunology; Infectious Disease TRD; BSL3 Core Facility, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

    Justin Jang Hann Chu

  2. The University of British Columbia, Vancouver, BC, Canada

    Bintou Ahmadou Ahidjo

  3. BSL3 Core Facility, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore

    Chee Keng Mok

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The views expressed in this chapter reflect the results of research conducted by the author and do not necessarily reflect the official policy or position of the Department of the Navy, Department of Defense, the United States Government, nor the Henry M. Jackson Foundation for the Advancement of Military Medicine, Inc.

This work was supported/funded by work unit number: A1436, DHP RDT&E supplemental COVID funding.

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Balinsky, C., Jani, V., Sun, P., Williams, M., Defang, G., Porter, K.R. (2022). Pseudovirus-Based Assays for the Measurement of Antibody-Mediated Neutralization of SARS-CoV-2. In: Chu, J.J.H., Ahidjo, B.A., Mok, C.K. (eds) SARS-CoV-2. Methods in Molecular Biology, vol 2452. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2111-0_21

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