A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels

Abstract Here we describe a homogeneous bioluminescent immunoassay based on the interaction between Fc-tagged SARS-CoV-2 Spike RBD and human ACE2, and its detection by secondary antibodies labeled with NanoLuc luciferase fragments LgBit and SmBit. The assay utility for the discovery of novel inhibit...

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Autores principales:	Juliano Alves, Laurie Engel, Renata de Vasconcelos Cabral, Eduardo L. Rodrigues, Liane de Jesus Ribeiro, Luiza M. Higa, Orlando da Costa Ferreira Júnior, Terezinha Marta P. P. Castiñeiras, Isabela de Carvalho Leitão, Amilcar Tanuri, Said A. Goueli, Hicham Zegzouti
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/896cb46d47394f0b9df32dc086b1c970
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spelling	oai:doaj.org-article:896cb46d47394f0b9df32dc086b1c9702021-12-02T18:02:06ZA bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels10.1038/s41598-021-97330-32045-2322https://doaj.org/article/896cb46d47394f0b9df32dc086b1c9702021-09-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-97330-3https://doaj.org/toc/2045-2322Abstract Here we describe a homogeneous bioluminescent immunoassay based on the interaction between Fc-tagged SARS-CoV-2 Spike RBD and human ACE2, and its detection by secondary antibodies labeled with NanoLuc luciferase fragments LgBit and SmBit. The assay utility for the discovery of novel inhibitors was demonstrated with a panel of anti-RBD antibodies, ACE2-derived miniproteins and soluble ACE2. Studying the effect of RBD mutations on ACE2 binding showed that the N501Y mutation increased RBD apparent affinity toward ACE2 tenfold that resulted in escaping inhibition by some anti-RBD antibodies. In contrast, while E484K mutation did not highly change the binding affinity, it still escaped antibody inhibition likely due to changes in the epitope recognized by the antibody. Also, neutralizing antibodies (NAbs) from COVID-19 positive samples from two distinct regions (USA and Brazil) were successfully detected and the results further suggest the persistence of NAbs for at least 6 months post symptom onset. Finally, sera from vaccinated individuals were tested for NAbs and showed varying neutralizing activity after first and second doses, suggesting the assay can be used to assess immunity of vaccinated populations. Our results demonstrate the broad utility and ease of use of this methodology both for drug discovery and clinical research applications.Juliano AlvesLaurie EngelRenata de Vasconcelos CabralEduardo L. RodriguesLiane de Jesus RibeiroLuiza M. HigaOrlando da Costa Ferreira JúniorTerezinha Marta P. P. CastiñeirasIsabela de Carvalho LeitãoAmilcar TanuriSaid A. GoueliHicham ZegzoutiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-15 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Juliano Alves Laurie Engel Renata de Vasconcelos Cabral Eduardo L. Rodrigues Liane de Jesus Ribeiro Luiza M. Higa Orlando da Costa Ferreira Júnior Terezinha Marta P. P. Castiñeiras Isabela de Carvalho Leitão Amilcar Tanuri Said A. Goueli Hicham Zegzouti A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels
description	Abstract Here we describe a homogeneous bioluminescent immunoassay based on the interaction between Fc-tagged SARS-CoV-2 Spike RBD and human ACE2, and its detection by secondary antibodies labeled with NanoLuc luciferase fragments LgBit and SmBit. The assay utility for the discovery of novel inhibitors was demonstrated with a panel of anti-RBD antibodies, ACE2-derived miniproteins and soluble ACE2. Studying the effect of RBD mutations on ACE2 binding showed that the N501Y mutation increased RBD apparent affinity toward ACE2 tenfold that resulted in escaping inhibition by some anti-RBD antibodies. In contrast, while E484K mutation did not highly change the binding affinity, it still escaped antibody inhibition likely due to changes in the epitope recognized by the antibody. Also, neutralizing antibodies (NAbs) from COVID-19 positive samples from two distinct regions (USA and Brazil) were successfully detected and the results further suggest the persistence of NAbs for at least 6 months post symptom onset. Finally, sera from vaccinated individuals were tested for NAbs and showed varying neutralizing activity after first and second doses, suggesting the assay can be used to assess immunity of vaccinated populations. Our results demonstrate the broad utility and ease of use of this methodology both for drug discovery and clinical research applications.
format	article
author	Juliano Alves Laurie Engel Renata de Vasconcelos Cabral Eduardo L. Rodrigues Liane de Jesus Ribeiro Luiza M. Higa Orlando da Costa Ferreira Júnior Terezinha Marta P. P. Castiñeiras Isabela de Carvalho Leitão Amilcar Tanuri Said A. Goueli Hicham Zegzouti
author_facet	Juliano Alves Laurie Engel Renata de Vasconcelos Cabral Eduardo L. Rodrigues Liane de Jesus Ribeiro Luiza M. Higa Orlando da Costa Ferreira Júnior Terezinha Marta P. P. Castiñeiras Isabela de Carvalho Leitão Amilcar Tanuri Said A. Goueli Hicham Zegzouti
author_sort	Juliano Alves
title	A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels
title_short	A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels
title_full	A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels
title_fullStr	A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels
title_full_unstemmed	A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels
title_sort	bioluminescent and homogeneous sars-cov-2 spike rbd and hace2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/896cb46d47394f0b9df32dc086b1c970
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A bioluminescent and homogeneous SARS-CoV-2 spike RBD and hACE2 interaction assay for antiviral screening and monitoring patient neutralizing antibody levels

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