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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.
  • Crawford KHD; Division of Basic Sciences and Computational Biology Program, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
  • Eguia R; Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA.
  • Dingens AS; Medical Scientist Training Program, University of Washington, Seattle, WA 98195, USA.
  • Loes AN; Division of Basic Sciences and Computational Biology Program, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
  • Malone KD; Division of Basic Sciences and Computational Biology Program, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
  • Wolf CR; Division of Basic Sciences and Computational Biology Program, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
  • Chu HY; Division of Basic Sciences and Computational Biology Program, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA.
  • Tortorici MA; Division of Allergy and Infectious Diseases, University of Washington, Seattle, WA 98195, USA.
  • Veesler D; Division of Allergy and Infectious Diseases, University of Washington, Seattle, WA 98195, USA.
  • Murphy M; Department of Biochemistry, University of Washington, Seattle, WA 98109, USA.
  • Pettie D; Institute Pasteur & CNRS UMR 3569, Unité de Virologie Structurale, 75015 Paris, France.
  • King NP; Department of Biochemistry, University of Washington, Seattle, WA 98109, USA.
  • Balazs AB; Institute for Protein Design, University of Washington, Seattle, WA 98195, USA.
  • Bloom JD; Institute for Protein Design, University of Washington, Seattle, WA 98195, USA.
Viruses ; 12(5)2020 05 06.
Article in English | MEDLINE | ID: covidwho-1389513
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Semantic information from SemMedBD (by NLM)
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8. M Protein, multiple myeloma PART_OF 2019 novel coronavirus
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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.
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Full text: Available Collection: International databases Database: MEDLINE Document Type: Article Main subject: Neutralization Tests / Spike Glycoprotein, Coronavirus / Antibodies, Viral Subject: Neutralization Tests / Spike Glycoprotein, Coronavirus / Antibodies, Viral Language: English Year: 2020

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Full text: Available Collection: International databases Database: MEDLINE Document Type: Article Main subject: Neutralization Tests / Spike Glycoprotein, Coronavirus / Antibodies, Viral Subject: Neutralization Tests / Spike Glycoprotein, Coronavirus / Antibodies, Viral Language: English Year: 2020
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