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New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis

The emergence of the new pathogen SARS-CoV-2 determined a rapid need for monoclonal antibodies (mAbs) to detect the virus in biological fluids as a rapid tool to identify infected individuals to be treated or quarantined. The majority of commercially available antigenic tests for SARS-CoV-2 rely on...

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Autores principales: Mariotti, Sabrina, Chiantore, Maria Vincenza, Teloni, Raffaela, Iacobino, Angelo, Capocefalo, Antonio, Michelini, Zuleika, Borghi, Martina, Baggieri, Melissa, Marchi, Antonella, Bucci, Paola, Gioacchini, Silvia, D’Amelio, Raffaele, Brouwer, Philip J. M., Sandini, Silvia, Acchioni, Chiara, Sgarbanti, Marco, Di Virgilio, Antonio, Grasso, Felicia, Cara, Andrea, Negri, Donatella, Magurano, Fabio, Di Bonito, Paola, Nisini, Roberto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953266/
https://www.ncbi.nlm.nih.gov/pubmed/36831149
http://dx.doi.org/10.3390/biomedicines11020610
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author Mariotti, Sabrina
Chiantore, Maria Vincenza
Teloni, Raffaela
Iacobino, Angelo
Capocefalo, Antonio
Michelini, Zuleika
Borghi, Martina
Baggieri, Melissa
Marchi, Antonella
Bucci, Paola
Gioacchini, Silvia
D’Amelio, Raffaele
Brouwer, Philip J. M.
Sandini, Silvia
Acchioni, Chiara
Sgarbanti, Marco
Di Virgilio, Antonio
Grasso, Felicia
Cara, Andrea
Negri, Donatella
Magurano, Fabio
Di Bonito, Paola
Nisini, Roberto
author_facet Mariotti, Sabrina
Chiantore, Maria Vincenza
Teloni, Raffaela
Iacobino, Angelo
Capocefalo, Antonio
Michelini, Zuleika
Borghi, Martina
Baggieri, Melissa
Marchi, Antonella
Bucci, Paola
Gioacchini, Silvia
D’Amelio, Raffaele
Brouwer, Philip J. M.
Sandini, Silvia
Acchioni, Chiara
Sgarbanti, Marco
Di Virgilio, Antonio
Grasso, Felicia
Cara, Andrea
Negri, Donatella
Magurano, Fabio
Di Bonito, Paola
Nisini, Roberto
author_sort Mariotti, Sabrina
collection PubMed
description The emergence of the new pathogen SARS-CoV-2 determined a rapid need for monoclonal antibodies (mAbs) to detect the virus in biological fluids as a rapid tool to identify infected individuals to be treated or quarantined. The majority of commercially available antigenic tests for SARS-CoV-2 rely on the detection of N antigen in biologic fluid using anti-N antibodies, and their capacity to specifically identify subjects infected by SARS-CoV-2 is questionable due to several structural analogies among the N proteins of different coronaviruses. In order to produce new specific antibodies, BALB/c mice were immunized three times at 20-day intervals with a recombinant spike (S) protein. The procedure used was highly efficient, and 40 different specific mAbs were isolated, purified and characterized, with 13 ultimately being selected for their specificity and lack of cross reactivity with other human coronaviruses. The specific epitopes recognized by the selected mAbs were identified through a peptide library and/or by recombinant fragments of the S protein. In particular, the selected mAbs recognized different linear epitopes along the S1, excluding the receptor binding domain, and along the S2 subunits of the S protein of SARS-CoV-2 and its major variants of concern. We identified combinations of anti-S mAbs suitable for use in ELISA or rapid diagnostic tests, with the highest sensitivity and specificity coming from proof-of-concept tests using recombinant antigens, SARS-CoV-2 or biological fluids from infected individuals, that represent important additional tools for the diagnosis of COVID-19.
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spelling pubmed-99532662023-02-25 New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis Mariotti, Sabrina Chiantore, Maria Vincenza Teloni, Raffaela Iacobino, Angelo Capocefalo, Antonio Michelini, Zuleika Borghi, Martina Baggieri, Melissa Marchi, Antonella Bucci, Paola Gioacchini, Silvia D’Amelio, Raffaele Brouwer, Philip J. M. Sandini, Silvia Acchioni, Chiara Sgarbanti, Marco Di Virgilio, Antonio Grasso, Felicia Cara, Andrea Negri, Donatella Magurano, Fabio Di Bonito, Paola Nisini, Roberto Biomedicines Article The emergence of the new pathogen SARS-CoV-2 determined a rapid need for monoclonal antibodies (mAbs) to detect the virus in biological fluids as a rapid tool to identify infected individuals to be treated or quarantined. The majority of commercially available antigenic tests for SARS-CoV-2 rely on the detection of N antigen in biologic fluid using anti-N antibodies, and their capacity to specifically identify subjects infected by SARS-CoV-2 is questionable due to several structural analogies among the N proteins of different coronaviruses. In order to produce new specific antibodies, BALB/c mice were immunized three times at 20-day intervals with a recombinant spike (S) protein. The procedure used was highly efficient, and 40 different specific mAbs were isolated, purified and characterized, with 13 ultimately being selected for their specificity and lack of cross reactivity with other human coronaviruses. The specific epitopes recognized by the selected mAbs were identified through a peptide library and/or by recombinant fragments of the S protein. In particular, the selected mAbs recognized different linear epitopes along the S1, excluding the receptor binding domain, and along the S2 subunits of the S protein of SARS-CoV-2 and its major variants of concern. We identified combinations of anti-S mAbs suitable for use in ELISA or rapid diagnostic tests, with the highest sensitivity and specificity coming from proof-of-concept tests using recombinant antigens, SARS-CoV-2 or biological fluids from infected individuals, that represent important additional tools for the diagnosis of COVID-19. MDPI 2023-02-18 /pmc/articles/PMC9953266/ /pubmed/36831149 http://dx.doi.org/10.3390/biomedicines11020610 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Mariotti, Sabrina
Chiantore, Maria Vincenza
Teloni, Raffaela
Iacobino, Angelo
Capocefalo, Antonio
Michelini, Zuleika
Borghi, Martina
Baggieri, Melissa
Marchi, Antonella
Bucci, Paola
Gioacchini, Silvia
D’Amelio, Raffaele
Brouwer, Philip J. M.
Sandini, Silvia
Acchioni, Chiara
Sgarbanti, Marco
Di Virgilio, Antonio
Grasso, Felicia
Cara, Andrea
Negri, Donatella
Magurano, Fabio
Di Bonito, Paola
Nisini, Roberto
New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis
title New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis
title_full New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis
title_fullStr New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis
title_full_unstemmed New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis
title_short New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis
title_sort new monoclonal antibodies specific for different epitopes of the spike protein of sars-cov-2 and its major variants: additional tools for a more specific covid-19 diagnosis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9953266/
https://www.ncbi.nlm.nih.gov/pubmed/36831149
http://dx.doi.org/10.3390/biomedicines11020610
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