Flow cytometry multiplexed method for the detection of neutralizing human antibodies to the native SARS-CoV-2 spike protein
Entity
UAM. Departamento de Biología Molecular; UAM. Departamento de MedicinaPublisher
EMBO Press; Wiley Open AccessDate
2021-02-17Citation
10.15252/emmm.202013549
EMBO Molecular Medicine 13.3 (2021): e13549
ISSN
1757-4676DOI
10.15252/emmm.202013549Funded by
This work was funded by intramural grant CSIC-COVID19-004: 202020E081 (to B.A.) and CSIC-COVID19-004: 202020E165 (to MF). L.H has been supported by an FPI fellowship from the Spanish Ministry of Science and Innovation. I.B. has been supported by an H2020-MSCA-ITN-2016 training network grant of the European Union (GA 721358)Editor's Version
https://doi.org/10.15252/emmm.202013549Subjects
flow cytometry; method; S protein; SARS-CoV-2; seropositivity; COVID-19; Biología y Biomedicina / Biología; MedicinaRights
© 2021 The AuthorsAbstract
A correct identification of seropositive individuals for the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is of paramount relevance to assess the degree of protection of a human population to present and future outbreaks of the COVID-19 pandemic. We describe here a sensitive and quantitative flow cytometry method using the cytometer-friendly non-adherent Jurkat T-cell line that stably expresses the full-length native spike “S” protein of SARS-CoV-2 and a truncated form of the human EGFR that serves a normalizing role. S protein and huEGFRt coding sequences are separated by a T2A self-cleaving sequence, allowing to accurately quantify the presence of anti-S immunoglobulins by calculating a score based on the ratio of fluorescence intensities obtained by double-staining with the test sera and anti-EGFR. The method allows to detect immune individuals regardless of the result of other serological tests or even repeated PCR monitoring. As examples of its use, we show that as much as 28% of the personnel working at the CBMSO in Madrid is already immune. Additionally, we show that anti-S antibodies with protective neutralizing activity are long-lasting and can be detected in sera 8 months after infection
Files in this item
Google Scholar:Horndler, Lydia
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Delgado, Pilar
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Abia, David
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Balabanov, Ivaylo
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Martínez-Fleta, Pedro
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Cornish, Georgina
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Llamas, Miguel A.
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Serrano-Villar, Sergio
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Sánchez Madrid, Francisco
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Fresno Escudero, Manuel
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van Santen, Hisse M.
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Alarcón, Balbino
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