Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-19

The COVID-19 pandemic has put global public health at high risk, rapidly spreading around the world. Although several COVID-19 vaccines are available for mass immunization, the world still urgently needs highly effective, reliable, cost-effective, and safe SARS-CoV-2 coronavirus vaccines, as well as...

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Autores principales:	Tarlan Mamedov, Damla Yuksel, Merve Ilgın, Irem Gürbüzaslan, Burcu Gulec, Gulshan Mammadova, Aykut Ozdarendeli, Hazel Yetiskin, Busra Kaplan, Shaikh Terkis Islam Pavel, Muhammet Ali Uygut, Gulnara Hasanova
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	COVID-19 SARS-CoV-2 nucleocapsid protein RBD of SARS-CoV-2 plant transient expression system Medicine R
Acceso en línea:	https://doaj.org/article/876db6d6a8484852be2252aa6989e44b
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spelling	oai:doaj.org-article:876db6d6a8484852be2252aa6989e44b2021-11-25T19:11:25ZProduction and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-1910.3390/vaccines91113372076-393Xhttps://doaj.org/article/876db6d6a8484852be2252aa6989e44b2021-11-01T00:00:00Zhttps://www.mdpi.com/2076-393X/9/11/1337https://doaj.org/toc/2076-393XThe COVID-19 pandemic has put global public health at high risk, rapidly spreading around the world. Although several COVID-19 vaccines are available for mass immunization, the world still urgently needs highly effective, reliable, cost-effective, and safe SARS-CoV-2 coronavirus vaccines, as well as antiviral and therapeutic drugs, to control the COVID-19 pandemic given the emerging variant strains of the virus. Recently, we successfully produced receptor-binding domain (RBD) variants in the <i>Nicotiana benthamiana</i> plant as promising vaccine candidates against COVID-19 and demonstrated that mice immunized with these antigens elicited a high titer of RBD-specific antibodies with potent neutralizing activity against SARS-CoV-2. In this study, we engineered the nucleocapsid (N) protein and co-expressed it with RBD of SARS-CoV-2 in <i>Nicotiana benthamiana</i> plant to produce an antigen cocktail. The purification yields were about 22 or 24 mg of pure protein/kg of plant biomass for N or N+RBD antigens, respectively. The purified plant produced N protein was recognized by N protein-specific monoclonal and polyclonal antibodies demonstrating specific reactivity of mAb to plant-produced N protein. In this study, for the first time, we report the co-expression of RBD with N protein to produce a cocktail antigen of SARS-CoV-2, which elicited high-titer antibodies with potent neutralizing activity against SARS-CoV-2. Thus, obtained data support that a plant-produced antigen cocktail, developed in this study, is a promising vaccine candidate against COVID-19.Tarlan MamedovDamla YukselMerve IlgınIrem GürbüzaslanBurcu GulecGulshan MammadovaAykut OzdarendeliHazel YetiskinBusra KaplanShaikh Terkis Islam PavelMuhammet Ali UygutGulnara HasanovaMDPI AGarticleCOVID-19SARS-CoV-2nucleocapsid proteinRBD of SARS-CoV-2planttransient expression systemMedicineRENVaccines, Vol 9, Iss 1337, p 1337 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	COVID-19 SARS-CoV-2 nucleocapsid protein RBD of SARS-CoV-2 plant transient expression system Medicine R
spellingShingle	COVID-19 SARS-CoV-2 nucleocapsid protein RBD of SARS-CoV-2 plant transient expression system Medicine R Tarlan Mamedov Damla Yuksel Merve Ilgın Irem Gürbüzaslan Burcu Gulec Gulshan Mammadova Aykut Ozdarendeli Hazel Yetiskin Busra Kaplan Shaikh Terkis Islam Pavel Muhammet Ali Uygut Gulnara Hasanova Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-19
description	The COVID-19 pandemic has put global public health at high risk, rapidly spreading around the world. Although several COVID-19 vaccines are available for mass immunization, the world still urgently needs highly effective, reliable, cost-effective, and safe SARS-CoV-2 coronavirus vaccines, as well as antiviral and therapeutic drugs, to control the COVID-19 pandemic given the emerging variant strains of the virus. Recently, we successfully produced receptor-binding domain (RBD) variants in the <i>Nicotiana benthamiana</i> plant as promising vaccine candidates against COVID-19 and demonstrated that mice immunized with these antigens elicited a high titer of RBD-specific antibodies with potent neutralizing activity against SARS-CoV-2. In this study, we engineered the nucleocapsid (N) protein and co-expressed it with RBD of SARS-CoV-2 in <i>Nicotiana benthamiana</i> plant to produce an antigen cocktail. The purification yields were about 22 or 24 mg of pure protein/kg of plant biomass for N or N+RBD antigens, respectively. The purified plant produced N protein was recognized by N protein-specific monoclonal and polyclonal antibodies demonstrating specific reactivity of mAb to plant-produced N protein. In this study, for the first time, we report the co-expression of RBD with N protein to produce a cocktail antigen of SARS-CoV-2, which elicited high-titer antibodies with potent neutralizing activity against SARS-CoV-2. Thus, obtained data support that a plant-produced antigen cocktail, developed in this study, is a promising vaccine candidate against COVID-19.
format	article
author	Tarlan Mamedov Damla Yuksel Merve Ilgın Irem Gürbüzaslan Burcu Gulec Gulshan Mammadova Aykut Ozdarendeli Hazel Yetiskin Busra Kaplan Shaikh Terkis Islam Pavel Muhammet Ali Uygut Gulnara Hasanova
author_facet	Tarlan Mamedov Damla Yuksel Merve Ilgın Irem Gürbüzaslan Burcu Gulec Gulshan Mammadova Aykut Ozdarendeli Hazel Yetiskin Busra Kaplan Shaikh Terkis Islam Pavel Muhammet Ali Uygut Gulnara Hasanova
author_sort	Tarlan Mamedov
title	Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-19
title_short	Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-19
title_full	Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-19
title_fullStr	Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-19
title_full_unstemmed	Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-19
title_sort	production and characterization of nucleocapsid and rbd cocktail antigens of sars-cov-2 in <i>nicotiana benthamiana</i> plant as a vaccine candidate against covid-19
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/876db6d6a8484852be2252aa6989e44b
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Production and Characterization of Nucleocapsid and RBD Cocktail Antigens of SARS-CoV-2 in <i>Nicotiana benthamiana</i> Plant as a Vaccine Candidate against COVID-19

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