A trimeric human angiotensin-converting enzyme 2 as an anti-SARS-CoV-2 agent

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1. Version published to 10.1038/s41594-020-00549-3

   Jan 11, 2021
2. 

   ScreenIT

   Sep 21, 2020

   SciScore for 10.1101/2020.09.18.301952: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   All the ACE2 variants were expressed in HEK 293F cells by transient transfection using Opti-MEM (Gibco-Thermo Fisher Scientific, Waltham, MA).	HEK 293F suggested: RRID:CVCL_6642)
   The expression construct was transiently transfected in HEK 293T cells using polyethylenimine (Polysciences, Inc, Warrington, PA).	HEK 293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   Briefly, HEK293 cells were transfected with pCMV-AT1R-IRES-Puro gene using …

   SciScore for 10.1101/2020.09.18.301952: (What is this?)

   Please note, not all rigor criteria are appropriate for all manuscripts.

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   All the ACE2 variants were expressed in HEK 293F cells by transient transfection using Opti-MEM (Gibco-Thermo Fisher Scientific, Waltham, MA).	HEK 293F suggested: RRID:CVCL_6642)
   The expression construct was transiently transfected in HEK 293T cells using polyethylenimine (Polysciences, Inc, Warrington, PA).	HEK 293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   Briefly, HEK293 cells were transfected with pCMV-AT1R-IRES-Puro gene using Lipofectamine 3000 reagent (Thermo Fisher Scientifics).	HEK293 suggested: None
   Neutralization of HIV-based pseudovirus containing a full-length SARS-CoV-2 S protein was measured using a single-round infection assay in HEK 293T/ACE2 target cells.	HEK 293T/ACE2 suggested: None
   Pseudotyped virus particles were produced in 293T/17 cells (ATCC) by co-transfection of a plasmid encoding codon-optimized SARS-CoV-2 full-length S containing G614, a packaging plasmid pCMV ΔR8.2 expressing HIV gag and pol, and a luciferase reporter plasmid pHR’ CMV-Luc.	293T/17 suggested: ATCC Cat# CRL-11268, RRID:CVCL_1926)
   The 293T cell line stably overexpressing the human ACE2 protein was created by the Farzan group at Scripps Research Institute.	293T suggested: NCBI_Iran Cat# C498, RRID:CVCL_0063)
   200 μl of each dilution or control were added to confluent monolayers of NR-596 Vero E6 cells in triplicate and incubated for 1 hour at 37°C and 5% CO2.	Vero E6 suggested: None
   Software and Algorithms
   Sentences	Resources
   Automated data collection was carried out using SerialEM version65 at a nominal magnification of 105,000× and the K3 detector in counting mode (calibrated pixel size, 0.825 Å) at a exposure rate of ~14.8 electrons per physical pixel per second.	SerialEM suggested: (SerialEM, RRID:SCR_017293)
   All density maps were corrected from the modulation transfer function of the K3 detector and then sharpened by applying a temperature factor that was estimated using post-processing in RELION.	RELION suggested: (RELION, RRID:SCR_016274)
   Several rounds of manual building were performed in Coot.	Coot suggested: (Coot, RRID:SCR_014222)
   Briefly, HEK293 cells were transfected with pCMV-AT1R-IRES-Puro gene using Lipofectamine 3000 reagent (Thermo Fisher Scientifics).	Thermo Fisher Scientifics suggested: None
   The half maximal inhibitory concentrations (IC50) were calculated using GraphPad Prism 8.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

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   Results 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).

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   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.

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   Results from TrialIdentifier: We found the following clinical trial numbers in your paper:
   

   Identifier	Status	Title
   NCT04335136	Completed	Recombinant Human Angiotensin-converting Enzyme 2 (rhACE2) a…
   NCT00886353	Completed	Safety and Tolerability Study of APN01 (Recombinant Human An…

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   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

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   Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on pages 27 and 29. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

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   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.

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   Read the original source
3. 

   Version published to 10.1101/2020.09.18.301952 on bioRxiv

   Sep 18, 2020