The development of Nanosota-1 as anti-SARS-CoV-2 nanobody drug candidates

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1. 

   Version published to 10.7554/elife.64815 on eLife

   Aug 2, 2021
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   ScreenIT

   Nov 18, 2020

   SciScore for 10.1101/2020.11.17.386532: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols of the University of Texas Medical Branch (protocol number 2007072) and of the University of Minnesota (protocol number 2009-38426A).
   Randomization	Four groups of hamsters (n=6 each randomly assigned) were treated with Nanosota-1C-Fc via intraperitoneal injection at one of the following time points and dosages: (1) 24 hours pre-challenge at 20 mg/kg body weight of hamsters; (2) 4 hours post-challenge at 20 mg/kg body weight of hamsters; (3) 4 hours post-challenge at 10 mg/kg body weight of hamsters.
   Blinding	not detected.
   Power Analysis	At a …

   SciScore for 10.1101/2020.11.17.386532: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All of the animals were handled according to approved institutional animal care and use committee (IACUC) protocols of the University of Texas Medical Branch (protocol number 2007072) and of the University of Minnesota (protocol number 2009-38426A).
   Randomization	Four groups of hamsters (n=6 each randomly assigned) were treated with Nanosota-1C-Fc via intraperitoneal injection at one of the following time points and dosages: (1) 24 hours pre-challenge at 20 mg/kg body weight of hamsters; (2) 4 hours post-challenge at 20 mg/kg body weight of hamsters; (3) 4 hours post-challenge at 10 mg/kg body weight of hamsters.
   Blinding	not detected.
   Power Analysis	At a sample size of 6 animals per group, G*Power analysis indicates that we can detect an effect size of 1.6 with a power of .80 (alpha = .
   Sex as a biological variable	Half-life of Nanosota-1 drugs in mice: Male C57BL/6 mice (3 to 4 weeks old) (Envigo) were intravenously injected (tail-vein) with Nanosota-1C or Nanosota-1C-Fc (100 µg in 100 µl PBS buffer).
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Briefly, ELISA plates were coated with recombinant SARS-CoV-2 RBD-His or RBD-Fc, and were then incubated sequentially with nanobody drugs, HRP-conjugated anti-llama antibody (1:5,000) (Sigma) or HRP-conjugated anti-human-Fc antibody (1:5,000) (Jackson ImmunoResearch).	RBD-Fc suggested: None anti-llama suggested: None anti-human-Fc suggested: None
   The samples were then subjected to SDS-PAGE and analyzed through Western blot using an anti-His antibody.	anti-His suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Cell lines, plasmids and virus: HEK293T cells (American Type Culture Collection) were cultured in Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 units/mL penicillin, and 100 µg/mL streptomycin (Life Technologies). ss320 E. coli (	HEK293T suggested: None
   Vero E6 cells (American Type Culture Collection) were grown in Eagle’s minimal essential medium (EMEM) supplemented with penicillin (100 units/ml), streptomycin (100 µg/ml), and 10% fetal bovine serum (FBS).	Vero E6 suggested: RRID:CVCL_XD71)
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Half-life of Nanosota-1 drugs in mice: Male C57BL/6 mice (3 to 4 weeks old) (Envigo) were intravenously injected (tail-vein) with Nanosota-1C or Nanosota-1C-Fc (100 µg in 100 µl PBS buffer).	C57BL/6 suggested: None
   Software and Algorithms
   Sentences	Resources
   At a sample size of 6 animals per group, G*Power analysis indicates that we can detect an effect size of 1.6 with a power of .80 (alpha = .	G*Power suggested: (G*Power, RRID:SCR_013726)

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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:

   • No conflict of interest statement was detected. If there are no conflicts, we encourage authors to explicit state so.
   • 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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3. 

   Version published to 10.1101/2020.11.17.386532v1 on bioRxiv

   Nov 17, 2020