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

Crawford, Katharine H. D.; Eguia, Rachel; Dingens, Adam S.; Loes, Andrea N.; Malone, Keara D.; Wolf, Caitlin R.; Chu, Helen Y.; Tortorici, M. Alejandra; Veesler, David; Murphy, Michael; Pettie, Deleah; King, Neil P.; Balazs, Alejandro B.; Bloom, Jesse D.

doi:10.3390/v12050513

articleVirusesMay 6, 2020GOLD OA

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

KHKatharine H. D. Crawford RERachel Eguia ASAdam S. Dingens ANAndrea N. Loes KDKeara D. Malone

University of Washington · Fred Hutch Cancer Center · +3 more institutions

PubMed

Indexed incrossrefdoajpubmed

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…

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Topics & keywords

Topics

Keywords

Virology
Neutralization
Vesicular stomatitis virus
Antibody
Neutralizing antibody
Virus
Biology
Coronavirus

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