Plant-Produced Glycosylated and In Vivo Deglycosylated Receptor Binding Domain Proteins of SARS-CoV-2 Induce Potent Neutralizing Responses in Mice

被引:25
作者
Mamedov, Tarlan [1 ]
Yuksel, Damla [1 ]
Ilgin, Merve [1 ]
Gurbuzaslan, Irem [1 ]
Gulec, Burcu [1 ]
Yetiskin, Hazel [2 ]
Uygut, Muhammet Ali [2 ]
Islam Pavel, Shaikh Terkis [2 ,3 ]
Ozdarendeli, Aykut [2 ,3 ]
Mammadova, Gulshan [1 ]
Say, Deniz [1 ]
Hasanova, Gulnara [1 ]
机构
[1] Akdeniz Univ, Dept Agr Biotechnol, TR-07058 Antalya, Turkey
[2] Erciyes Univ, Fac Med, Dept Microbiol, TR-38280 Kayseri, Turkey
[3] Erciyes Univ, Dev & Applicat Ctr, Vaccine Res, TR-38280 Kayseri, Turkey
来源
VIRUSES-BASEL | 2021年 / 13卷 / 08期
关键词
COVID-19; SARS-CoV-2; spike protein; receptor binding domain (RBD) vaccine; plant; transient expression system; SPIKE PROTEIN; IMMUNE-RESPONSES; SARS-COV; GLYCOPROTEIN; MUTATIONS; INFECTIONS; EXPRESSION; VACCINES; VIRUS; MERS;
D O I
10.3390/v13081595
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
The COVID-19 pandemic, caused by SARS-CoV-2, has rapidly spread to more than 222 countries and has put global public health at high risk. The world urgently needs cost-effective and safe SARS-CoV-2 vaccines, antiviral, and therapeutic drugs to control it. In this study, we engineered the receptor binding domain (RBD) of the SARS-CoV-2 spike (S) protein and produced it in the plant Nicotiana benthamiana in a glycosylated and deglycosylated form. Expression levels of both glycosylated (gRBD) and deglycosylated (dRBD) RBD were greater than 45 mg/kg fresh weight. The purification yields were 22 mg of pure protein/kg of plant biomass for gRBD and 20 mg for dRBD, which would be sufficient for commercialization of these vaccine candidates. The purified plant-produced RBD protein was recognized by an S protein-specific monoclonal antibody, demonstrating specific reactivity of the antibody to the plant-produced RBD proteins. The SARS-CoV-2 RBD showed specific binding to angiotensin converting enzyme 2 (ACE2), the SARS-CoV-2 receptor. In mice, the plant-produced RBD antigens elicited high titers of antibodies with a potent virus-neutralizing activity. To our knowledge, this is the first report demonstrating that mice immunized with plant-produced deglycosylated RBD form elicited high titer of RBD-specific antibodies with potent neutralizing activity against SARS-CoV-2 infection. Thus, obtained data support that plant-produced glycosylated and in vivo deglycosylated RBD antigens, developed in this study, are promising vaccine candidates for the prevention of COVID-19.
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页数:19
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