Protocol to isolate and assess spike protein cleavage in SARS-CoV-2 variants obtained from clinical COVID-19 samples

For efficient cell entry, SARS-CoV-2 spike protein needs to be cleaved by cellular proteases. Here, we present a comprehensive protocol to assess SARS-CoV-2 spike protein cleavage in viral supernatants from SARS-CoV-2-infected cells. We also include a previous step of SARS-CoV-2 isolation from nasop...

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Autores principales: Escalera, Alba, García-Sastre, Adolfo, Aydillo, Teresa
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2022
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9189002/
https://www.ncbi.nlm.nih.gov/pubmed/35776647
http://dx.doi.org/10.1016/j.xpro.2022.101502
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author Escalera, Alba
García-Sastre, Adolfo
Aydillo, Teresa
author_facet Escalera, Alba
García-Sastre, Adolfo
Aydillo, Teresa
author_sort Escalera, Alba
collection PubMed
description For efficient cell entry, SARS-CoV-2 spike protein needs to be cleaved by cellular proteases. Here, we present a comprehensive protocol to assess SARS-CoV-2 spike protein cleavage in viral supernatants from SARS-CoV-2-infected cells. We also include a previous step of SARS-CoV-2 isolation from nasopharyngeal swabs of patients with COVID-19. We optimized the procedures to enhance successful viral isolation and specific spike detection. This protocol facilitates the evaluation of the role of spike mutations in spike protein processing. For complete details on the use and execution of this protocol, please refer to Escalera et al. (2022).
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spelling pubmed-91890022022-06-13 Protocol to isolate and assess spike protein cleavage in SARS-CoV-2 variants obtained from clinical COVID-19 samples Escalera, Alba García-Sastre, Adolfo Aydillo, Teresa STAR Protoc Protocol For efficient cell entry, SARS-CoV-2 spike protein needs to be cleaved by cellular proteases. Here, we present a comprehensive protocol to assess SARS-CoV-2 spike protein cleavage in viral supernatants from SARS-CoV-2-infected cells. We also include a previous step of SARS-CoV-2 isolation from nasopharyngeal swabs of patients with COVID-19. We optimized the procedures to enhance successful viral isolation and specific spike detection. This protocol facilitates the evaluation of the role of spike mutations in spike protein processing. For complete details on the use and execution of this protocol, please refer to Escalera et al. (2022). Elsevier 2022-06-13 /pmc/articles/PMC9189002/ /pubmed/35776647 http://dx.doi.org/10.1016/j.xpro.2022.101502 Text en © 2022 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Escalera, Alba
García-Sastre, Adolfo
Aydillo, Teresa
Protocol to isolate and assess spike protein cleavage in SARS-CoV-2 variants obtained from clinical COVID-19 samples
title Protocol to isolate and assess spike protein cleavage in SARS-CoV-2 variants obtained from clinical COVID-19 samples
title_full Protocol to isolate and assess spike protein cleavage in SARS-CoV-2 variants obtained from clinical COVID-19 samples
title_fullStr Protocol to isolate and assess spike protein cleavage in SARS-CoV-2 variants obtained from clinical COVID-19 samples
title_full_unstemmed Protocol to isolate and assess spike protein cleavage in SARS-CoV-2 variants obtained from clinical COVID-19 samples
title_short Protocol to isolate and assess spike protein cleavage in SARS-CoV-2 variants obtained from clinical COVID-19 samples
title_sort protocol to isolate and assess spike protein cleavage in sars-cov-2 variants obtained from clinical covid-19 samples
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9189002/
https://www.ncbi.nlm.nih.gov/pubmed/35776647
http://dx.doi.org/10.1016/j.xpro.2022.101502
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