Rapid selection of a human monoclonal antibody that potently neutralizes SARS-CoV-2 in two animal models

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   ScreenIT

   Jun 8, 2020

   SciScore for 10.1101/2020.05.13.093088: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: The study was carried out in accordance with the recommendations for care and use of animals by the Office of Laboratory Animal Welfare (OLAW), National Institutes of Health and the Institutional Animal Care and Use Committee (IACUC) of University of North Carolina (UNC permit no. A-3410-01).
   Randomization	The Fab and scFv libraries were constructed by randomly pairing antibody VH and VL gene, and the VH libraries -by grafting CDRs into stable VH scaffolds.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Groups of 5 each of 10 to 12-month old female BALB/c mice (Envigo, #047) were treated prophylactically (12 hours before …

   SciScore for 10.1101/2020.05.13.093088: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: The study was carried out in accordance with the recommendations for care and use of animals by the Office of Laboratory Animal Welfare (OLAW), National Institutes of Health and the Institutional Animal Care and Use Committee (IACUC) of University of North Carolina (UNC permit no. A-3410-01).
   Randomization	The Fab and scFv libraries were constructed by randomly pairing antibody VH and VL gene, and the VH libraries -by grafting CDRs into stable VH scaffolds.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Groups of 5 each of 10 to 12-month old female BALB/c mice (Envigo, #047) were treated prophylactically (12 hours before infection) intraperitoneally with 900 μg, 200 μg, or 50 μg of IgG1 ab1, respectively.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Generation, Expression and Characterization of SARS-CoV-2 RBD-Fc, S1-Fc, ACE2-Fc and CR3022 Fab: The SARS-CoV-2 surface glycoprotein and the anti-SARS-CoV antibody IgG1 CR3022 (31) and IgG1 S230 genes were synthesized by IDT (Coralville, Iowa).	S1-Fc suggested: None ACE2-Fc suggested: None SARS-CoV-2 surface glycoprotein suggested: None anti-SARS-CoV suggested: None IgG1 suggested: None
   MERS-CoV-specific IgG1 m336 antibody was expressed in human mammalian cell as described previously (5).	MERS-CoV-specific IgG1 suggested: None
   After washing, bound IgG1 CR3022 was detected by HRP-conjugated anti human Fc antibody.	anti human Fc antibody. suggested: None
   Then cells were resuspended to 100 μl PBSA buffer followed by addition of 1 μl PE conjugated anti-human Fc antibody (Sigma-Aldrich) and incubate at 4 °C for 30 min.	anti-human Fc suggested: None
   FcγRs expression level was FACS tested by using FITC conjugated anti-FcγR antibody (Invitrogen).	anti-FcγR suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Briefly, HIV-1 backbone based pseudovirus was packaged in 293T cells by co-transfecting with plasmid encoding SARS-CoV-2 S protein and plasmid encoding luciferase expressing HIV-1 genome (pNL4-3.luc.RE) using polyethylenimine (PEI).	293T suggested: None
   Additionally, Vero E6 cells treated with the MERS-CoV RBD-specific neutralizing m336 mAb (5) were included as additional controls.	Vero E6 suggested: None
   The entire library of plasmids is arrayed in duplicate in a matrix format and transfected into HEK-293T cells, followed by incubation for 36 h to allow protein expression.	HEK-293T suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Groups of 5 each of 10 to 12-month old female BALB/c mice (Envigo, #047) were treated prophylactically (12 hours before infection) intraperitoneally with 900 μg, 200 μg, or 50 μg of IgG1 ab1, respectively.	BALB/c suggested: None
   Ethics statement: Human ACE2 transgenic C3B6 mice (6–9 weeks old) and BALB/c mice (10-12 weeks old) were used for all experiments.	C3B6 suggested: None
   Software and Algorithms
   Sentences	Resources
   The concentrations at which IgG1 ab1 or hACE2-Fc achieved 50% PE-A+ cells (EC50) was obtained by fitting using the equation of “[agonist] vs. response -- variable slope (four parameter)” in the Graphpad Prism 7 (San Diego, CA).	Graphpad suggested: (GraphPad, RRID:SCR_000306)
   Statistical analyses: The statistical significance of difference between IgG1 treated and control mice lung virus titers was analyzed by the one-tailed Mann Whitney U test calculated using GraphPad Prism 7.0.	GraphPad Prism 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: 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.
   • 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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   Read the original source
2. 

   Version published to 10.1101/2020.05.13.093088v2 on bioRxiv

   Jun 2, 2020
3. 

   Version published to 10.1101/2020.05.13.093088v1 on bioRxiv

   May 14, 2020