Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent of coronavirus disease (COVID-19) which has recently emerged as a potential threat to global public health. SARS-CoV-2 is the third known human coronavirus that has huge impact on the human population after SARS-CoV and MERS-CoV. Although some vaccines and therapeutic drugs are currently in clinical trials, none of them are approved for commercial use yet. As with SARS-CoV, SARS-CoV-2 utilizes angiotensin-converting enzyme 2 (ACE2) as the cell entry receptor to enter into the host cell. In this study, we have transiently produced human ACE2 fused with the Fc region of human IgG1 in Nicotiana benthamiana and the in vitro neutralization efficacy of the plant-produced ACE2-Fc fusion protein was assessed. The recombinant ACE2-Fc fusion protein was expressed in N. benthamiana at 100 μg/g leaf fresh weight on day 6 post-infiltration. The recombinant fusion protein showed potent binding to receptor binding domain (RBD) of SARS-CoV-2. Importantly, the plant-produced fusion protein exhibited potent anti-SARS-CoV-2 activity in vitro. Treatment with ACE2-Fc fusion protein after viral infection dramatically inhibit SARS-CoV-2 infectivity in Vero cells with an IC50 value of 0.84 μg/ml. Moreover, treatment with ACE2-Fc fusion protein at the pre-entry stage suppressed SARS-CoV-2 infection with an IC50 of 94.66 μg/ml. These findings put a spotlight on the plant-produced ACE2-Fc fusion protein as a potential therapeutic candidate against SARS-CoV-2.
Highlights
The coronavirus disease 2019 (COVID-19) outbreak originated in Wuhan, China in late December 2019 (Han et al, 2020; Li et al, 2020; Lupia et al, 2020)
COVID-19 is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which belongs to Coronaviridae family
The angiotensin-converting enzyme 2 (ACE2)-Fc fusion protein gene was designed using codon preferred for N. benthamiana and commercially synthesized
Summary
The coronavirus disease 2019 (COVID-19) outbreak originated in Wuhan, China in late December 2019 (Han et al, 2020; Li et al, 2020; Lupia et al, 2020). The outbreak has spread to more than 200 countries with more than 53.7 million confirmed cases and more than 1.3 million confirmed deaths as of 17 November 2020 (World Health Organization, 2020b). These numbers are still increasing with the ongoing pandemic which overwhelms national health care systems and has had major consequences on global economy. COVID-19 is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which belongs to Coronaviridae family. Several members of the family Coronaviridae constantly circulate in the human population and usually cause mild respiratory disease (Ciotti et al, 2020; Han et al, 2020). The closely related severe acute respiratory syndrome coronavirus (SARS-CoV) and the Middle East respiratory syndrome coronavirus (MERS-CoV) are initially transmitted from animals to humans and cause severe respiratory diseases (Fehr et al, 2017)
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