Virus‐Free and Live‐Cell Visualizing SARS‐CoV‐2 Cell Entry for Studies of Neutralizing Antibodies and Compound Inhibitors

Article activity feed

1. Version published to 10.1002/smtd.202001031

   Dec 18, 2020
2. 

   ScreenIT

   Jul 25, 2020

   SciScore for 10.1101/2020.07.22.215236: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: The study was approved by the institutional review board of the School of Public Health in accordance with the Declaration of Helsinki, and written informed consent was obtained. Consent: The study was approved by the institutional review board of the School of Public Health in accordance with the Declaration of Helsinki, and written informed consent was obtained.
   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

   Antibodies
   Sentences	Resources
   Generation and production of antibodies against SARS-CoV-2 S: Balb/c mice were intraperitoneal …

   SciScore for 10.1101/2020.07.22.215236: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IRB: The study was approved by the institutional review board of the School of Public Health in accordance with the Declaration of Helsinki, and written informed consent was obtained. Consent: The study was approved by the institutional review board of the School of Public Health in accordance with the Declaration of Helsinki, and written informed consent was obtained.
   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

   Antibodies
   Sentences	Resources
   Generation and production of antibodies against SARS-CoV-2 S: Balb/c mice were intraperitoneal immunized with 5 μg of SARS-CoV2-RBD (expression in this study, n=5), SARS-CoV2-S1 (Sino Biological, 40591-V08H, n=3) and SARS-CoV2-S2 (Sino Biological, 40590-V08B, n=3), respectively.	SARS-CoV2-S2 suggested: None
   The titers for TAb, IgG, and IgM antibody were calculated via S/CO multiplied by the maximum dilution factors.	IgM suggested: None
   Commercial antibodies were used to detect intracellular ACE2 (Sino Biological, 10108-T56), TMPRSS2 (Abcam, ab92323), and GAPDH (Proteintech, HRP-60004) according to the manufacturer’s instructions.	TMPRSS2 suggested: (Abcam Cat# ab92323, RRID:AB_10585592) GAPDH suggested: (Proteintech Cat# HRP-60004, RRID:AB_2737588)
   Experimental Models: Cell Lines
   Sentences	Resources
   Cell lines: The cell lines of 293T, H1299, H1299-ACE2hR, 293T-ACE2hR and 293T-ACE2iRb3 were Dulbecco’s modified Eagle medium (Sigma, D6429) supplemented with 10% fetal bovine serum (Thermo Scientific, 10099-141), 0.1 mM non-essential amino acids (Thermo Scientific,1140-050), and were incubated at 37C° and 5% CO2 in a humidified incubator.	H1299 suggested: None
   The mixtures were then added to a monolayer of Vero cells (104 cells per well, pre-washed twice with PBS) in a 96-well plate and incubated at 37°C.	Vero suggested: RRID:CVCL_ZW93)
   Cell imaging assays: For direct visualizing the cellular binding and uptake of RBD or spike proteins, the 293T-ACE2iRb3 cells were seeded at 2×104 cells per well in poly-D-lysine pretreated CellCarrier-96 Black plate.	293T-ACE2iRb3 suggested: None
   To visualize compound-induced influence on viral entry, 293T-ACE2iRb3 (Figure 6C-D, Figure S11A) or H1299-ACE2hR cells (Figure S11B) were pretreated with serial dilutions of compounds for 1-hour.	H1299-ACE2hR suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Generation and production of antibodies against SARS-CoV-2 S: Balb/c mice were intraperitoneal immunized with 5 μg of SARS-CoV2-RBD (expression in this study, n=5), SARS-CoV2-S1 (Sino Biological, 40591-V08H, n=3) and SARS-CoV2-S2 (Sino Biological, 40590-V08B, n=3), respectively.	Balb/c suggested: None
   Software and Algorithms
   Sentences	Resources
   Particle picking, two rounds of reference-free 2D classification and final 3D reconstruction were performed by the programs cryoSPARC v2 42.	cryoSPARC suggested: (cryoSPARC, RRID:SCR_016501)
   The ID50/IC50 values were determined by 4PL regression GraphPad Prism v8.0.	GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   GraphPad Prism version 8.0.1 was used for all statistical calculations.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

   ---

   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).

   ---

   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: No clinical trial numbers were referenced.

   ---

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

   ---

   Results from JetFighter: Please consider improving the rainbow (“jet”) colormap(s) used on page 33. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.

   ---

   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.

   ---

   Read the original source
3. 

   Version published to 10.1101/2020.07.22.215236 on bioRxiv

   Jul 22, 2020