The dual function monoclonal antibodies VIR-7831 and VIR-7832 demonstrate potent in vitro and in vivo activity against SARS-CoV-2

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   ScreenIT

   Mar 15, 2021

   SciScore for 10.1101/2021.03.09.434607: (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	Twelve- to sixteen-week-old male hamsters were interperitoneally administered a non-LS version of VIR-7831 (SGHmAb-no-LS), control antibody or diluent Day -1 or Day -2 prior to virus challenge.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Monoclonal Antibodies: VIR-7831 and VIR-7832 were produced at WuXi Biologics (China).	VIR-7832 suggested: None
   Spike binding affinity quantification by SPR: Antibody was diluted to 2 μg/mL (1 mL) in HBS-EP+ buffer and injected at 10 μL/min for 30 seconds across one flow cell of a …

   SciScore for 10.1101/2021.03.09.434607: (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	Twelve- to sixteen-week-old male hamsters were interperitoneally administered a non-LS version of VIR-7831 (SGHmAb-no-LS), control antibody or diluent Day -1 or Day -2 prior to virus challenge.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Monoclonal Antibodies: VIR-7831 and VIR-7832 were produced at WuXi Biologics (China).	VIR-7832 suggested: None
   Spike binding affinity quantification by SPR: Antibody was diluted to 2 μg/mL (1 mL) in HBS-EP+ buffer and injected at 10 μL/min for 30 seconds across one flow cell of a CM5 sensor chip immobilized with anti-human Fc antibody docked in a Biacore T200.	anti-human Fc suggested: None
   SARS-CoV2-RBD diluted in HBS-EP+ buffer was then injected at a single concentration, 1:3 dilutions from 100 nM to 3.7 nM, across both the flow cell containing captured the antibody as well as a reference flow cell containing only anti-human Fc antibody.	anti-human Fc antibody. suggested: None
   After an overnight incubation at 37°C, monocytes were stained with anti-human CD14-APC antibody (BD Pharmingen).	anti-human CD14-APC suggested: None
   Plates were washed three times with PBS and then incubated for one hour at RT with 50 μL/well of goat anti-rabbit-Alexa647 secondary antibody at 1:1,000 in blocking buffer.	anti-rabbit-Alexa647 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Pseudotyped virus production: Lenti-X™ 293T cells were seeded in 10-cm dishes for 80% next day confluency.	293T suggested: None
   Live virus neutralization: VeroE6 cells were seeded into flat bottom 96-well plates at 20,000 cells/well and cultured overnight at 37°C.	VeroE6 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Jurkat effector cells expressing indicated FcγRs and stably transfected with an NFAT-driven luciferase gene were thawed, diluted in assay buffer, and added to the plate at an effector to target cell ratio of 6:1 for FcRγIIIa and FcγRIIb or 5:1 for FcγIIa.	Jurkat suggested: TKG Cat# TKG 0209, RRID:CVCL_0065)
   On the day of infection, moDCs, PBMCs, and U937 cells were counted and plated at 7.5×104 cells/well.	U937 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Binding to Cell Surface Expressed SARS-CoV-2 Spike Protein: The SARS-CoV-2 spike protein coding sequence (YP_009724390.1, Wuhan-Hu-1 strain) was cloned into a cell expression plasmid under the control of the human CMV promoter (phCMV1) to generate phCMV1 WT spike.	Wuhan-Hu-1 suggested: None
   Software and Algorithms
   Sentences	Resources
   Data were analyzed and graphed using GraphPad Prism software (v8.4.1).	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   Raw data were exported from the flow cytometer into the flow cytometry analysis software FlowJo v10 (Becton Dickinson) for gating and determination of the percentage of CD14+ cells that were also double positive for cell trace violet and PKH67.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   The percent of nucleocapsid+ cells was quantified using the Gen5 Imager software (Biotek, Vermont) as number of Cy5+ cells, [(nucleocapsid+ cells)/number of Hoechst+ cells (total cells)]×100.	Gen5 Imager suggested: None
   Data was analyzed using Prism v8.00 (GraphPad Software, La Jolla California USA, www.graphpad.com).	Prism suggested: (PRISM, RRID:SCR_005375) GraphPad suggested: (GraphPad Prism, RRID:SCR_002798)
   The spike open reading frame was localized by aligning the reference protein sequence (NCBI reference sequence: YP_009724390.1) to the genomic sequence of isolates with Exonerate v.2.4.0.	Exonerate suggested: (Exonerate, RRID:SCR_016088)
   Multiple sequence alignment was performed using MAFFT v.7.455.	MAFFT suggested: (MAFFT, RRID:SCR_011811)
   3.10 package Biostrings v.2.54.0.	Biostrings suggested: (Biostrings, RRID:SCR_016949)

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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 found bar graphs of continuous data. We recommend replacing bar graphs with more informative graphics, as many different datasets can lead to the same bar graph. The actual data may suggest different conclusions from the summary statistics. For more information, please see Weissgerber et al (2015).

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   Results from JetFighter: We did not find any issues relating to colormaps.

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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 10.1101/2021.03.09.434607 on bioRxiv

   Mar 10, 2021