High level production and characterization of truncated human angiotensin converting enzyme 2 in Nicotiana benthamiana plant as a potential therapeutic target in COVID-19
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Abstract
The COVID-19 pandemic, which is 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 safe, a cost-effective SARS-CoV-2 coronavirus vaccine, therapeutic and antiviral drugs to combat the COVID-19. Angiotensin-converting enzyme 2 (ACE2), as a key receptor for SARS-CoV-2 infections, has been proposed as a potential therapeutic target in COVID-19 patients. In this study, we report high level production (about ∼0.75 g /kg leaf biomass) of glycosylated and non-glycosylated forms of recombinant human truncated ACE2 in Nicotiana benthamiana plant. The plant produced recombinant human truncated ACE2s successfully bind to the SARC-CoV-2 spike protein, but deglycosylated ACE2 binds more strongly than the glycosylated counterpart. Importantly, both deglycosylated and glycosylated forms of AEC2 stable at elevated temperatures for prolonged periods and demonstrated strong anti-SARS-CoV-2 activity in vitro . The IC 50 values of glycosylated and deglycosylated AEC2 were 0.4 and 24 μg/ml, respectively, for the pre-entry infection, when incubated with 100TCID 50 of SARS-CoV-2. Thus, plant produced truncated ACE2s are promising cost-effective and safe candidate as a potential therapeutic targets in the treatment of COVID-19 patients.
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SciScore for 10.1101/2021.05.17.444533: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After washing, wells were incubated with anti-mouse HRP -IgG antibody (BioLegend). anti-mouse HRP -IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources A total of 150 µL incubated mixture was then inoculated on Vero E6 Cells grown in a 96-well flat-bottomed tissue culture plate (Greiner, Germany). Vero E6suggested: NoneRecombinant DNA Sentences Resources AGL1 carrying the pEAQ-ACE2 plasmid was then infiltrated into 6-7-week old N. benthamiana plants. pEAQ-ACE2suggested: None… SciScore for 10.1101/2021.05.17.444533: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics not detected. Sex as a biological variable not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources After washing, wells were incubated with anti-mouse HRP -IgG antibody (BioLegend). anti-mouse HRP -IgGsuggested: NoneExperimental Models: Cell Lines Sentences Resources A total of 150 µL incubated mixture was then inoculated on Vero E6 Cells grown in a 96-well flat-bottomed tissue culture plate (Greiner, Germany). Vero E6suggested: NoneRecombinant DNA Sentences Resources AGL1 carrying the pEAQ-ACE2 plasmid was then infiltrated into 6-7-week old N. benthamiana plants. pEAQ-ACE2suggested: NoneSoftware and Algorithms Sentences Resources Elution fractions were collected as 0.5 ml/eppendorf and protein concentrations in the eluted fractions were measured by BioDrop. BioDropsuggested: NoneResults from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.Results from TrialIdentifier: We found the following clinical trial numbers in your paper:
Identifier Status Title NCT00886353 Completed Safety and Tolerability Study of APN01 (Recombinant Human An… NCT01597635 Completed The Safety, Tolerability, PK and PD of GSK2586881 in Patient… Results from Barzooka: We did not find any issues relating to the usage of bar graphs.
Results from JetFighter: We did not find any issues relating to colormaps.
Results from rtransparent:- Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
- Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
- No protocol registration statement was detected.
Results from scite Reference Check: We found no unreliable references.
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