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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.<\/jats:p>","DOI":"10.3390\/v12050513","type":"journal-article","created":{"date-parts":[[2020,5,7]],"date-time":"2020-05-07T03:10:38Z","timestamp":1588821038000},"page":"513","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":725,"title":["Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6223-4019","authenticated-orcid":false,"given":"Katharine H. 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Alejandra","family":"Tortorici","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98109, USA"},{"name":"Institute Pasteur &amp; CNRS UMR 3569, Unit\u00e9 de Virologie Structurale, Paris 75015, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Veesler","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98109, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8438-2653","authenticated-orcid":false,"given":"Michael","family":"Murphy","sequence":"additional","affiliation":[{"name":"Institute for Protein Design, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5477-7195","authenticated-orcid":false,"given":"Deleah","family":"Pettie","sequence":"additional","affiliation":[{"name":"Institute for Protein Design, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2978-4692","authenticated-orcid":false,"given":"Neil P.","family":"King","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Washington, Seattle, WA 98109, USA"},{"name":"Institute for Protein Design, University of Washington, Seattle, WA 98195, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1767-3944","authenticated-orcid":false,"given":"Alejandro B.","family":"Balazs","sequence":"additional","affiliation":[{"name":"The Ragon Institute of Massachusetts General Hospital, the Massachusetts Institute Technology, and Harvard University, Cambridge, MA 02139, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1267-3408","authenticated-orcid":false,"given":"Jesse D.","family":"Bloom","sequence":"additional","affiliation":[{"name":"Division of Basic Sciences and Computational Biology Program, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA"},{"name":"Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA"},{"name":"Howard Hughes Medical Institute, Seattle, WA 98103, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,5,6]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Ju, B., Zhang, Q., Ge, X., Wang, R., Yu, J., Shan, S., Zhou, B., Song, S., Tang, X., and Yu, J. 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