Humanized Single Domain Antibodies Neutralize SARS-CoV-2 by Targeting Spike Receptor Binding Domain

Article activity feed

1. 

   ScreenIT

   Jun 5, 2020

   SciScore for 10.1101/2020.04.14.042010: (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

   Antibodies
   Sentences	Resources
   Antibodies were obtained from ThermoFisher for anti-His-HRP, anti-human IgG-HRP, anti-His-488 and anti-CM13.	anti-His-HRP suggested: None anti-human IgG-HRP suggested: None anti-His-488 suggested: None
   Library design and construction: A synthetic sdAb phage display library was used for the screening of SARS-CoV-2 neutralizing antibodies.	SARS-CoV-2 neutralizing antibodies . suggested: None
   After 4 rounds of panning, phage ELISA identification was performed with 480 …

   SciScore for 10.1101/2020.04.14.042010: (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

   Antibodies
   Sentences	Resources
   Antibodies were obtained from ThermoFisher for anti-His-HRP, anti-human IgG-HRP, anti-His-488 and anti-CM13.	anti-His-HRP suggested: None anti-human IgG-HRP suggested: None anti-His-488 suggested: None
   Library design and construction: A synthetic sdAb phage display library was used for the screening of SARS-CoV-2 neutralizing antibodies.	SARS-CoV-2 neutralizing antibodies . suggested: None
   After 4 rounds of panning, phage ELISA identification was performed with 480 individual colonies using Anti-CM13 antibody [B62-FE2] (HRP).	Anti-CM13 suggested: None
   Maxipreped plasmids were transiently transfected into 293-F cells (Thermofisher) and the cells were further cultured in suspension for 6 days before harvesting antibody-containing supernatant.	antibody-containing supernatant . suggested: None
   The primary antibody used for Western blot was a horseradish peroxidase conjugated goat anti-mouse IgG (Sigma-Aldrich, St. Louis, MO, USA)	anti-mouse IgG suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Cells and reagents: The Vero (African green monkey kidney), HEK293T (human kidney epithelial), 293F, Calu-3 (human lung adenocarcinoma) cells were obtained from China Infrastructure of Cell Line Resource (Beijing, China) and maintained in Dulbecco’s modified Eagle’s medium (DMEM, ThermoFisher, Waltham, MA, USA) supplemented with 2-10% fetal bovine serum (FBS,	Calu-3 suggested: None
   Production of SARS-CoV-2 spike pseudotyped particle (SARS-CoV-2pp) and virus entry assay: To produce SARS-CoV-2pp, HEK293T cells were seeded 1 day prior to transfection at 2.5×106 cells in a 10-cm plate.	HEK293T suggested: None
   To conduct the virus entry assay, Vero E6 or other cells were seeded in a 96-well plate at 1 day prior to transduction.	Vero E6 suggested: None
   For antibody neutralization assay, Vero cells were seeded in 96-well plates at 1 day prior to infection.	Vero suggested: None
   Immunofluorescence microscopy and Western blot: Cultured 293T cells on coverslips were transfected with either SARS-CoV-2 S expression plasmid or empty vector for 24 h and then fixed using 4% paraformaldehyde for 15 min at room temperature, permeabilized with 0.1% Triton X-100 (Sigma-Aldrich) in PBS for 10 min.	293T suggested: None
   Software and Algorithms
   Sentences	Resources
   Statistical analysis: Data were analyzed using GraphPad Prism 6.01 (GraphPad Software, San Diego, CA, USA).	GraphPad 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: 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.

   ---

   Read the original source
2. 

   Version published to 10.1101/2020.04.14.042010 on bioRxiv

   Apr 15, 2020