IgA MAb blocks SARS-CoV-2 Spike-ACE2 interaction providing mucosal immunity

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1. 

   ScreenIT

   Jun 8, 2020

   SciScore for 10.1101/2020.05.15.096719: (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
   Isotype switching was conducted using primers designed to amplify the variable heavy chain of the IgG antibody.	IgG suggested: None
   IgG and IgA1 antibodies were transfected in Expi293 cells and purified as previously described32.	IgA1 suggested: None
   After incubation, the cell pellets were washed and then resuspened in PBS with 2% FBS and incubated with 10 µg/mL of anti-Myc antibody for 1 hour at 4°C.	anti-Myc suggested: None
   Experimental Models: Cell Lines
   Sentence…

   SciScore for 10.1101/2020.05.15.096719: (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
   Isotype switching was conducted using primers designed to amplify the variable heavy chain of the IgG antibody.	IgG suggested: None
   IgG and IgA1 antibodies were transfected in Expi293 cells and purified as previously described32.	IgA1 suggested: None
   After incubation, the cell pellets were washed and then resuspened in PBS with 2% FBS and incubated with 10 µg/mL of anti-Myc antibody for 1 hour at 4°C.	anti-Myc suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   IgG and IgA1 antibodies were transfected in Expi293 cells and purified as previously described32.	Expi293 suggested: RRID:CVCL_D615)
   Flow cytometry-based receptor binding inhibition assay: Vero E6 cells were harvested with PBS containing 5 mM EDTA and aliquoted to 1×106 cells per reaction.	Vero E6 suggested: None
   Before mixing with the cells, Myc-tagged SARS-CoV S1-590 or SARS-CoV-2 S1-604 was incubated with the MAb at varying concentrations for 1 hour at room temperature, then the S protein was added to the Vero cells to a final concentration of 10 nM.	Vero suggested: None
   SARS-S and SARS2-S spike protein was provided in trans by co-transfection of 293T cells with pcDNA-G with pNL4-3.	293T suggested: None
   Software and Algorithms
   Sentences	Resources
   Cells were washed twice then subjected to flow cytometric analysis using a MACSquant Flow Cytometer (Miltenyi Biotec) and analyzed by FlowJo (version 10).	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   The molecular modeling of MAb362 was performed through the program Modeller 9.15 using the basic modeling and forming the initial MAb362:SARS-CoV RBD complex.	Modeller suggested: (MODELLER, RRID:SCR_008395)
   Hydrogen bonds were determined for pairs of eligible donor/acceptor atoms using criteria set by Schrodinger (Schrodinger, LLC, The PyMOL Molecular Graphics System, Version 1.3r1. 2010.).	PyMOL suggested: (PyMOL, RRID:SCR_000305)
   Each assay was performed in triplicate. c Statistical Analysis: Statistical calculations were performed using Prism version 7.03 (GraphPad Software, La Jolla, CA).	Prism suggested: (PRISM, RRID:SCR_005375) GraphPad 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: No clinical trial numbers were referenced.

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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 page 14. 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.
   • No funding statement was detected.
   • No protocol registration statement was detected.

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2. 

   Version published to 10.1101/2020.05.15.096719v1 on bioRxiv

   May 15, 2020