Therapeutic activity of an inhaled potent SARS-CoV-2 neutralizing human monoclonal antibody in hamsters

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1. Version published to 10.1016/j.xcrm.2021.100218

   Mar 1, 2021
2. 

   ScreenIT

   Oct 15, 2020

   SciScore for 10.1101/2020.10.14.339150: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	Consent: The subjects provided signed written informed consent. IRB: All procedures and methods were approved by the Institutional Review Board for Human Use at the University of Alabama at Birmingham, and all experiments were performed in accordance with relevant guidelines and regulations.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	In a prophylactic experiment, 6-week old, female hamsters were IP injection of 10 mg/kg of hmAb 1212C2 (n=6), 1206D1 (n=3), human IgG1 Isotype control mAb (BioXcell, Labanon, NH; n=6), or PBS (n=5).
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resour…

   SciScore for 10.1101/2020.10.14.339150: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	Consent: The subjects provided signed written informed consent. IRB: All procedures and methods were approved by the Institutional Review Board for Human Use at the University of Alabama at Birmingham, and all experiments were performed in accordance with relevant guidelines and regulations.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	In a prophylactic experiment, 6-week old, female hamsters were IP injection of 10 mg/kg of hmAb 1212C2 (n=6), 1206D1 (n=3), human IgG1 Isotype control mAb (BioXcell, Labanon, NH; n=6), or PBS (n=5).
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   At 24 hpi, infected cells were fixed with 10% neutral formalin for 24 h and were immune-stained using anti-NP monoclonal 1C7C7 antibody (21).	anti-NP suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Paired heavy and light chain genes cloned into IgG1 expression vectors and were transfected into HEK293T cells and culture supernatant was concentrated using 100,000 MWCO Amicon Ultra centrifugal filters (Millipore-Sigma, Cork, Ireland), and IgG captured and eluted from Magne Protein A beads (Promega, Madison, WI) as previously described (27, 39).	HEK293T suggested: None
   Nucleocapsid (Sino Biological), and HepG2 whole cell lysate (Abcam, Cambridge, MA)	HepG2 suggested: CLS Cat# 300198/p2277_Hep-G2, RRID:CVCL_0027)
   Briefly, confluent monolayers of Vero E6 cells were mock infected or infected with the indicated SARS-CoV-2.	Vero E6 suggested: RRID:CVCL_XD71)
   VeroE6/TMPRSS2 cells were seeded at 2 x 104 cells/well in opaque plates (Greiner 655083).	VeroE6/TMPRSS2 suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
   Software and Algorithms
   Sentences	Resources
   Cryopreserved cells were thawed and stained for flow cytometry similar as previously described (39), using anti-CD 19-APC-Cy7 (SJ25C1, BD Biosciences), HIV gp140-AlexaFluor647, SARS-CoV-2 RBD-BV421, CD3-BV510 (OKT3, Biolegend), CD4-BV510 (HI30, Biolegend), IgD-FITC (IA6-2, BD Biosciences), CD27-PE (CLB-27/1, Life Technologies), Annexin V-PerCP-Cy5.5 (Biolegend), and Live/Dead aqua (Molecular	BD Biosciences suggested: (BD Biosciences, RRID:SCR_013311)
   Immunoglobulin sequences were analyzed by IgBlast (www.ncbi.nlm.nih.gov/igblast) and IMGT/V-QUEST (http://www.imgt.org/IMGT_vquest/vquest) to determine which sequences should lead to productive immunoglobulin, to identify the germline V(D)J gene segments with the highest identity, and to scrutinize sequence properties.	IgBlast suggested: (IgBLAST, RRID:SCR_002873)
   A non-linear regression curve fit analysis over the dilution curve can be performed using GraphPad Prism to calculate NT50.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   For each day, lungs and nasal turbinate were collected removed and photographed on white paper towels to look for gross pathology using a macroscopic pathology scoring analysis measure the distributions of pathological lesions, including consolidation, congestion, and pneumonic lesions using ImageJ software (NIH).	ImageJ suggested: (ImageJ, RRID:SCR_003070)

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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: Please consider improving the rainbow (“jet”) colormap(s) used on page 38. 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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3. 

   Version published to 10.1101/2020.10.14.339150v1 on bioRxiv

   Oct 14, 2020