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A cell-based system combined with flow cytometry to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in patients with COVID-19
This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing H...
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| Published in: | STAR protocols 2022-03, Vol.3 (1), p.101229-101229, Article 101229 |
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| container_end_page | 101229 |
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| container_title | STAR protocols |
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| creator | Martin, Sophie Jégou, Gwénaële Nicolas, Aurore Le Gallo, Matthieu Chevet, Éric Godey, Florence Avril, Tony |
| description | This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness.
For complete details on the use and execution of this protocol, please refer to Martin et al. (2021).
[Display omitted]
•A human-cell-based assay was developed to access antibody responses against SARS-CoV-2•The assay recapitulates spike membrane insertion and posttranslational modifications•The assay represents a powerful tool to test spike variants and vaccine effectiveness
This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness. |
| doi_str_mv | 10.1016/j.xpro.2022.101229 |
| format | article |
| fullrecord | <record><control><sourceid>proquest_doaj_</sourceid><recordid>TN_cdi_doaj_primary_oai_doaj_org_article_faf35d5423bf4e3b8ed989ac576f1f86</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><els_id>S2666166722001095</els_id><doaj_id>oai_doaj_org_article_faf35d5423bf4e3b8ed989ac576f1f86</doaj_id><sourcerecordid>2639229112</sourcerecordid><originalsourceid>FETCH-LOGICAL-c506t-ef769a695c21f58e9a9a86cdc89047a3375c5b9a3211a0db6fd86cdd71d332793</originalsourceid><addsrcrecordid>eNp9Uk1vEzEQXSEQrUr_AAfkIxw2-CPrXUsIKUqBRopUiUKvltcep45218F2UvI7-MN42VK1HDiNPfPmzdcritcEzwgm_P129nMX_IxiSkcHpeJZcUo55yXhvH7-6H1SnMe4xRjTitA5aV4WJ6yiTU25OC1-LZCGritbFcGgeIwJeqR937oh_-9cukW283dIH5PvIYUjSh7BQXV7lQCpIbnWmyMKEHd-iBCR2ig3xISuF1-vy6W_KSlKQQ2xh77NFlDuOkGGIDegnUoOhhSnQsurm9VFScSr4oVVXYTze3tWfP_86dvyslxffVktF-tSV5inEmzNheKi0pTYqgGhhGq4NroReF4rxupKV61QjBKisGm5NWPY1MQwRmvBzorVxGu82spdcL0KR-mVk38cPmykCsnpDqRVllWmmlPW2jmwtgEjGqF0VXNLbMMz18eJa7dvezA6TxVU94T0aWRwt3LjD7JpOK7x2My7ieD2n7TLxVqOPsw4o2yODyRj394XC_7HHmKSvYvjGfN-_T5KypnIgiCEZiidoDr4GAPYB26C5SgkuZWjkOQoJDkJKSe9eTzMQ8pf2WTAhwkA-TwHB0FGnQ-pwbgAOuX9uf_x_wa569uQ</addsrcrecordid><sourcetype>Open Website</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>2639229112</pqid></control><display><type>article</type><title>A cell-based system combined with flow cytometry to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in patients with COVID-19</title><source>ScienceDirect®</source><source>PubMed Central</source><creator>Martin, Sophie ; Jégou, Gwénaële ; Nicolas, Aurore ; Le Gallo, Matthieu ; Chevet, Éric ; Godey, Florence ; Avril, Tony</creator><creatorcontrib>Martin, Sophie ; Jégou, Gwénaële ; Nicolas, Aurore ; Le Gallo, Matthieu ; Chevet, Éric ; Godey, Florence ; Avril, Tony</creatorcontrib><description>This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness.
For complete details on the use and execution of this protocol, please refer to Martin et al. (2021).
[Display omitted]
•A human-cell-based assay was developed to access antibody responses against SARS-CoV-2•The assay recapitulates spike membrane insertion and posttranslational modifications•The assay represents a powerful tool to test spike variants and vaccine effectiveness
This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness.</description><identifier>ISSN: 2666-1667</identifier><identifier>EISSN: 2666-1667</identifier><identifier>DOI: 10.1016/j.xpro.2022.101229</identifier><identifier>PMID: 35287269</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Antibody ; Antibody Formation ; Cancer ; Cell Biology ; Cell-based Assays ; COVID-19 ; Flow Cytometry - methods ; Flow Cytometry/Mass Cytometry ; Health Sciences ; HEK293 Cells ; Humans ; Life Sciences ; Membrane Proteins ; Microbiology ; Molecular Biology ; Protocol ; SARS-CoV-2 ; Spike Glycoprotein, Coronavirus</subject><ispartof>STAR protocols, 2022-03, Vol.3 (1), p.101229-101229, Article 101229</ispartof><rights>2022 The Author(s)</rights><rights>2022 The Author(s).</rights><rights>Attribution - NonCommercial - NoDerivatives</rights><rights>2022 The Author(s) 2022</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><cites>FETCH-LOGICAL-c506t-ef769a695c21f58e9a9a86cdc89047a3375c5b9a3211a0db6fd86cdd71d332793</cites><orcidid>0000-0002-1129-8514 ; 0000-0002-1256-3674</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8860709/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S2666166722001095$$EHTML$$P50$$Gelsevier$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,3548,27923,27924,45779,53790,53792</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35287269$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink><backlink>$$Uhttps://hal.science/hal-03632340$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Martin, Sophie</creatorcontrib><creatorcontrib>Jégou, Gwénaële</creatorcontrib><creatorcontrib>Nicolas, Aurore</creatorcontrib><creatorcontrib>Le Gallo, Matthieu</creatorcontrib><creatorcontrib>Chevet, Éric</creatorcontrib><creatorcontrib>Godey, Florence</creatorcontrib><creatorcontrib>Avril, Tony</creatorcontrib><title>A cell-based system combined with flow cytometry to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in patients with COVID-19</title><title>STAR protocols</title><addtitle>STAR Protoc</addtitle><description>This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness.
For complete details on the use and execution of this protocol, please refer to Martin et al. (2021).
[Display omitted]
•A human-cell-based assay was developed to access antibody responses against SARS-CoV-2•The assay recapitulates spike membrane insertion and posttranslational modifications•The assay represents a powerful tool to test spike variants and vaccine effectiveness
This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness.</description><subject>Antibody</subject><subject>Antibody Formation</subject><subject>Cancer</subject><subject>Cell Biology</subject><subject>Cell-based Assays</subject><subject>COVID-19</subject><subject>Flow Cytometry - methods</subject><subject>Flow Cytometry/Mass Cytometry</subject><subject>Health Sciences</subject><subject>HEK293 Cells</subject><subject>Humans</subject><subject>Life Sciences</subject><subject>Membrane Proteins</subject><subject>Microbiology</subject><subject>Molecular Biology</subject><subject>Protocol</subject><subject>SARS-CoV-2</subject><subject>Spike Glycoprotein, Coronavirus</subject><issn>2666-1667</issn><issn>2666-1667</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>DOA</sourceid><recordid>eNp9Uk1vEzEQXSEQrUr_AAfkIxw2-CPrXUsIKUqBRopUiUKvltcep45218F2UvI7-MN42VK1HDiNPfPmzdcritcEzwgm_P129nMX_IxiSkcHpeJZcUo55yXhvH7-6H1SnMe4xRjTitA5aV4WJ6yiTU25OC1-LZCGritbFcGgeIwJeqR937oh_-9cukW283dIH5PvIYUjSh7BQXV7lQCpIbnWmyMKEHd-iBCR2ig3xISuF1-vy6W_KSlKQQ2xh77NFlDuOkGGIDegnUoOhhSnQsurm9VFScSr4oVVXYTze3tWfP_86dvyslxffVktF-tSV5inEmzNheKi0pTYqgGhhGq4NroReF4rxupKV61QjBKisGm5NWPY1MQwRmvBzorVxGu82spdcL0KR-mVk38cPmykCsnpDqRVllWmmlPW2jmwtgEjGqF0VXNLbMMz18eJa7dvezA6TxVU94T0aWRwt3LjD7JpOK7x2My7ieD2n7TLxVqOPsw4o2yODyRj394XC_7HHmKSvYvjGfN-_T5KypnIgiCEZiidoDr4GAPYB26C5SgkuZWjkOQoJDkJKSe9eTzMQ8pf2WTAhwkA-TwHB0FGnQ-pwbgAOuX9uf_x_wa569uQ</recordid><startdate>20220318</startdate><enddate>20220318</enddate><creator>Martin, Sophie</creator><creator>Jégou, Gwénaële</creator><creator>Nicolas, Aurore</creator><creator>Le Gallo, Matthieu</creator><creator>Chevet, Éric</creator><creator>Godey, Florence</creator><creator>Avril, Tony</creator><general>Elsevier Inc</general><general>Elsevier</general><scope>6I.</scope><scope>AAFTH</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope><scope>1XC</scope><scope>VOOES</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-1129-8514</orcidid><orcidid>https://orcid.org/0000-0002-1256-3674</orcidid></search><sort><creationdate>20220318</creationdate><title>A cell-based system combined with flow cytometry to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in patients with COVID-19</title><author>Martin, Sophie ; Jégou, Gwénaële ; Nicolas, Aurore ; Le Gallo, Matthieu ; Chevet, Éric ; Godey, Florence ; Avril, Tony</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c506t-ef769a695c21f58e9a9a86cdc89047a3375c5b9a3211a0db6fd86cdd71d332793</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Antibody</topic><topic>Antibody Formation</topic><topic>Cancer</topic><topic>Cell Biology</topic><topic>Cell-based Assays</topic><topic>COVID-19</topic><topic>Flow Cytometry - methods</topic><topic>Flow Cytometry/Mass Cytometry</topic><topic>Health Sciences</topic><topic>HEK293 Cells</topic><topic>Humans</topic><topic>Life Sciences</topic><topic>Membrane Proteins</topic><topic>Microbiology</topic><topic>Molecular Biology</topic><topic>Protocol</topic><topic>SARS-CoV-2</topic><topic>Spike Glycoprotein, Coronavirus</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Martin, Sophie</creatorcontrib><creatorcontrib>Jégou, Gwénaële</creatorcontrib><creatorcontrib>Nicolas, Aurore</creatorcontrib><creatorcontrib>Le Gallo, Matthieu</creatorcontrib><creatorcontrib>Chevet, Éric</creatorcontrib><creatorcontrib>Godey, Florence</creatorcontrib><creatorcontrib>Avril, Tony</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><collection>Hyper Article en Ligne (HAL)</collection><collection>Hyper Article en Ligne (HAL) (Open Access)</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>STAR protocols</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Martin, Sophie</au><au>Jégou, Gwénaële</au><au>Nicolas, Aurore</au><au>Le Gallo, Matthieu</au><au>Chevet, Éric</au><au>Godey, Florence</au><au>Avril, Tony</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A cell-based system combined with flow cytometry to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in patients with COVID-19</atitle><jtitle>STAR protocols</jtitle><addtitle>STAR Protoc</addtitle><date>2022-03-18</date><risdate>2022</risdate><volume>3</volume><issue>1</issue><spage>101229</spage><epage>101229</epage><pages>101229-101229</pages><artnum>101229</artnum><issn>2666-1667</issn><eissn>2666-1667</eissn><abstract>This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness.
For complete details on the use and execution of this protocol, please refer to Martin et al. (2021).
[Display omitted]
•A human-cell-based assay was developed to access antibody responses against SARS-CoV-2•The assay recapitulates spike membrane insertion and posttranslational modifications•The assay represents a powerful tool to test spike variants and vaccine effectiveness
This protocol describes a flow cytometry approach to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in COVID-19-positive patient sera samples without the need of specific laboratory facilities for viral infection. We developed a human-cell-based system using spike-expressing HEK293T cells that mimics membrane insertion and N-glycosylation of viral integral membrane proteins in host cells. This assay represents a powerful tool to test antibody responses against SARS-CoV-2 variants and vaccine effectiveness.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>35287269</pmid><doi>10.1016/j.xpro.2022.101229</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0002-1129-8514</orcidid><orcidid>https://orcid.org/0000-0002-1256-3674</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | STAR protocols, 2022-03, Vol.3 (1), p.101229-101229, Article 101229 |
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| language | eng |
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| source | ScienceDirect®; PubMed Central |
| subjects | Antibody Antibody Formation Cancer Cell Biology Cell-based Assays COVID-19 Flow Cytometry - methods Flow Cytometry/Mass Cytometry Health Sciences HEK293 Cells Humans Life Sciences Membrane Proteins Microbiology Molecular Biology Protocol SARS-CoV-2 Spike Glycoprotein, Coronavirus |
| title | A cell-based system combined with flow cytometry to evaluate antibody responses against SARS-CoV-2 transmembrane proteins in patients with COVID-19 |
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