Isolation and Characterization of Mouse Monoclonal Antibodies That Neutralize SARS-CoV-2 and Its Variants of Concern Alpha, Beta, Gamma and Delta by Binding Conformational Epitopes of Glycosylated RBD With High Potency
| العنوان: | Isolation and Characterization of Mouse Monoclonal Antibodies That Neutralize SARS-CoV-2 and Its Variants of Concern Alpha, Beta, Gamma and Delta by Binding Conformational Epitopes of Glycosylated RBD With High Potency |
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| المؤلفون: | Maria Laura De Angelis, Antonella Marchi, Sabrina Mariotti, Valeria Esposito, Paola Di Bonito, Chiara Acchioni, Maria Franca Pirillo, Giulietta Venturi, Maria Vincenza Chiantore, Paul F. McKay, Fabio Tosini, Alessandra Gallinaro, Donatella R.M. Negri, Francesco Marino, Marco Sgarbanti, Andrea Canitano, Fabio Magurano, Roberto Nisini, Antonio Di Virgilio, Antonio Capocefalo, Paola Bucci, Andrea Cara, Angelo Iacobino, Melissa Baggieri, Silvia Sandini, Martina Borghi, Zuleika Michelini, Raffaela Teloni |
| المصدر: | Frontiers in Immunology, Vol 12 (2021) Frontiers in Immunology |
| بيانات النشر: | Frontiers Media S.A., 2021. |
| سنة النشر: | 2021 |
| مصطلحات موضوعية: | Glycosylation, Immunoprecipitation, medicine.drug_class, diagnosis, Immunology, Antibodies, Viral, Monoclonal antibody, Epitope, law.invention, Epitopes, Western blot, Neutralization Tests, law, Cell Line, Tumor, Chlorocebus aethiops, medicine, Animals, Humans, Immunology and Allergy, Vero Cells, Original Research, chemistry.chemical_classification, SARS-COV-2 variants, Mice, Inbred BALB C, therapy, biology, medicine.diagnostic_test, SARS-CoV-2, pathogenesis, epitopes expression, Antibodies, Monoclonal, RC581-607, Antibodies, Neutralizing, Virology, COVID-19 Drug Treatment, HEK293 Cells, chemistry, Spike Glycoprotein, Coronavirus, Vero cell, Recombinant DNA, biology.protein, Female, Angiotensin-Converting Enzyme 2, Binding Sites, Antibody, Antibody, neutralizing monoclonal antibodies, Immunologic diseases. Allergy, Glycoprotein |
| الوصف: | Antibodies targeting Receptor Binding Domain (RBD) of SARS-CoV-2 have been suggested to account for the majority of neutralizing activity in COVID-19 convalescent sera and several neutralizing antibodies (nAbs) have been isolated, characterized and proposed as emergency therapeutics in the form of monoclonal antibodies (mAbs). However, SARS-CoV-2 variants are rapidly spreading worldwide from the sites of initial identification. The variants of concern (VOC) B.1.1.7 (Alpha), B.1.351 (Beta), P.1 (Gamma) and B.1.167.2 (Delta) showed mutations in the SARS-CoV-2 spike protein potentially able to cause escape from nAb responses with a consequent reduction of efficacy of vaccines and mAbs-based therapy. We produced the recombinant RBD (rRBD) of SARS-CoV-2 spike glycoprotein from the Wuhan-Hu 1 reference sequence in a mammalian system, for mice immunization to isolate new mAbs with neutralizing activity. Here we describe four mAbs that were able to bind the rRBD in Enzyme-Linked Immunosorbent Assay and the transmembrane full-length spike protein expressed in HEK293T cells by flow cytometry assay. Moreover, the mAbs recognized the RBD in supernatants of SARS-CoV-2 infected VERO E6 cells by Western Blot under non-reducing condition or in supernatants of cells infected with lentivirus pseudotyped for spike protein, by immunoprecipitation assay. Three out of four mAbs lost their binding efficiency to completely N-deglycosylated rRBD and none was able to bind the same recombinant protein expressed in Escherichia coli, suggesting that the epitopes recognized by three mAbs are generated by the conformational structure of the glycosylated native protein. Of particular relevance, three mAbs were able to inhibit Wuhan SARS-CoV-2 infection of VERO E6 cells in a plaque-reduction neutralization test and the Wuhan SARS-CoV-2 as well as the Alpha, Beta, Gamma and Delta VOC in a pseudoviruses-based neutralization test. These mAbs represent important additional tools for diagnosis and therapy of COVID-19 and may contribute to the understanding of the functional structure of SARS-CoV-2 RBD. |
| اللغة: | English |
| تدمد: | 1664-3224 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ac5bee35b866fd6815b5be8acaf4225c https://www.frontiersin.org/articles/10.3389/fimmu.2021.750386/full |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....ac5bee35b866fd6815b5be8acaf4225c |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 16643224 |
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