Double Lock of a Potent Human Monoclonal Antibody against SARS-CoV-2

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   ScreenIT

   Nov 26, 2020

   SciScore for 10.1101/2020.11.24.393629: (What is this?)

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

   Table 1: Rigor

   NIH rigor criteria are not applicable to paper type.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Generation of humanized anti-SARS-CoV-2 antibody HB27: SARS-CoV-2 antibodies were screened from a phage-display scFv library constructed from the spleen mRNA of mice immunized with recombinant SARS-CoV-2 RBD protein.	anti-SARS-CoV-2 suggested: None
   SARS-CoV-2 RBD was used as the bait to select for specific anti-RBD scFvs by biopanning and the scFvs exhibiting potent binding for SARS-CoV-2 RBD were generated as chimeric antibodies.	anti-RBD suggested: None
   HB27 antibody was incubated for 1h, followed by incubation of RBD-His and anti-His-PE,or APC labelled ACE2-Fc for 20 min.	anti-His-PE suggested: (Miltenyi …

   SciScore for 10.1101/2020.11.24.393629: (What is this?)

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

   Table 1: Rigor

   NIH rigor criteria are not applicable to paper type.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Generation of humanized anti-SARS-CoV-2 antibody HB27: SARS-CoV-2 antibodies were screened from a phage-display scFv library constructed from the spleen mRNA of mice immunized with recombinant SARS-CoV-2 RBD protein.	anti-SARS-CoV-2 suggested: None
   SARS-CoV-2 RBD was used as the bait to select for specific anti-RBD scFvs by biopanning and the scFvs exhibiting potent binding for SARS-CoV-2 RBD were generated as chimeric antibodies.	anti-RBD suggested: None
   HB27 antibody was incubated for 1h, followed by incubation of RBD-His and anti-His-PE,or APC labelled ACE2-Fc for 20 min.	anti-His-PE suggested: (Miltenyi Biotec Cat# 130-092-691, RRID:AB_1103227)
   Then the virus was mixed with 0.3 μM ACE2 and HB27 antibody with the final concentration of 2.5 μM, 0.5 μM, 0.1 μM or 0.02 μM.	HB27 suggested: None
   Five fields were automatically collected in each well to count the number of fused and unfused cells and the antibody inhibition rate was calculated as following: fusion rate (FR) = (fused cell number) / (fused cell number+ unfused cell number), Inhibition%= (Positive Control (FR) – Sample (FR)) / (Positive Control (FR)) %.	FR suggested: None FR ) – Sample ( FR) ) / ( Positive Control ( FR suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Polyethylenimine was used to transiently transfect HEK Expi 293F cells (Thermo Fisher) with SARS-CoV RBD, SARS-CoV-2 RBD and SARS-CoV-2 S, respectively.	HEK Expi 293F suggested: None
   60 μL of SARS-CoV/SARS-CoV-2 pseudoviruses and 60 μL of serial diluted antibody samples were incubated for 1 h at 37°C, after which the pseudovirus-mAb mixtures were added into the wells containing Vero-E6 cells.	Vero-E6 suggested: None
   Immunofluorescence: 293T cells were transfected with SARS-CoV-2-S-GFP or ACE2-GFP.	293T suggested: None
   Plaque reduction neutralization tests (PRNT): The neutralization activity of HB27 against SARS-CoV-2 were examined by standard plaque reduction neutralization tests (PRNT) in Vero cells.	Vero suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Briefly, a group of 6 to 8-week-old hACE2 humanized mice or BALB/c mice were intraperitoneally administrated with HB27 (20 mg/kg) before (prophylactic) and/or after (therapeutic) challenge with 5 × 105 PFU of SARS-CoV-2 or 1.6×104 PFU of MASCp6 via intranasal route, respectively.	BALB/c suggested: None
   Software and Algorithms
   Sentences	Resources
   Data were analyzed using Flowjo and Graphpad.	Flowjo suggested: (FlowJo, RRID:SCR_008520) Graphpad suggested: (GraphPad, RRID:SCR_000306)
   Movies (32 frames, each 0.2 s, total dose 60 e−Å−2) were recorded at defocuses of between 1.25 and 2.7 μm using SerialEM, yielding a final pixel size of 1.05 Å.	SerialEM suggested: (SerialEM, RRID:SCR_017293)
   Model building and refinement: The structures of SARS-CoV-2 S trimer and a human Fab fragment (Protein Data Bank ID: 6VSB and 5N4J, respectively) were manually fitted into the refined map of SARS-CoV-2 S trimer-HB27 complex in Chimera (Pettersen et al., 2004) and then improved by manual real-space refinement in COOT (Brown et al., 2015).	COOT suggested: (Coot, RRID:SCR_014222)
   The final models were evaluated using Molprobity (Chen et al., 2010).	Molprobity suggested: (MolProbity, RRID:SCR_014226)
   Briefly, a group of 6 to 8-week-old hACE2 humanized mice or BALB/c mice were intraperitoneally administrated with HB27 (20 mg/kg) before (prophylactic) and/or after (therapeutic) challenge with 5 × 105 PFU of SARS-CoV-2 or 1.6×104 PFU of MASCp6 via intranasal route, respectively.	SARS-CoV-2 suggested: (Active Motif Cat# 91351, RRID:AB_2847848)

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   Results from OddPub: Thank you for sharing your data.

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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
   NCT04483375	Completed	Safety, Tolerability and Pharmacokinetics of SCTA01, an Anti…

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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 40, 41 and 21. 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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   Version published to 10.1101/2020.11.24.393629v1 on bioRxiv

   Nov 25, 2020