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Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays

Authors :
Katharine H. D. Crawford
Rachel Eguia
Adam S. Dingens
Andrea N. Loes
Keara D. Malone
Caitlin R. Wolf
Helen Y. Chu
M. Alejandra Tortorici
David Veesler
Michael Murphy
Deleah Pettie
Neil P. King
Alejandro B. Balazs
Jesse D. Bloom
Source :
Viruses, Vol 12, Iss 5, p 513 (2020)
Publication Year :
2020
Publisher :
MDPI AG, 2020.

Abstract

SARS-CoV-2 enters cells using its Spike protein, which is also the main target of neutralizing antibodies. Therefore, assays to measure how antibodies and sera affect Spike-mediated viral infection are important for studying immunity. Because SARS-CoV-2 is a biosafety-level-3 virus, one way to simplify such assays is to pseudotype biosafety-level-2 viral particles with Spike. Such pseudotyping has now been described for single-cycle lentiviral, retroviral, and vesicular stomatitis virus (VSV) particles, but the reagents and protocols are not widely available. Here, we detailed how to effectively pseudotype lentiviral particles with SARS-CoV-2 Spike and infect 293T cells engineered to express the SARS-CoV-2 receptor, ACE2. We also made all the key experimental reagents available in the BEI Resources repository of ATCC and the NIH. Furthermore, we demonstrated how these pseudotyped lentiviral particles could be used to measure the neutralizing activity of human sera or plasma against SARS-CoV-2 in convenient luciferase-based assays, thereby providing a valuable complement to ELISA-based methods that measure antibody binding rather than neutralization.

Details

Language :
English
ISSN :
19994915
Volume :
12
Issue :
5
Database :
Directory of Open Access Journals
Journal :
Viruses
Publication Type :
Academic Journal
Accession number :
edsdoj.88a2dcbba5204bbe892d526add0ceb42
Document Type :
article
Full Text :
https://doi.org/10.3390/v12050513