Recombinant Fc-fusion vaccine of RBD induced protection against SARS-CoV-2 in non-human primate and mice

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   ScreenIT

   Dec 1, 2020

   SciScore for 10.1101/2020.11.29.402339: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All animal procedures were reviewed and approved by the Animal Experiment Committee of Laboratory Animal Center, Academy of Military Medical Sciences (AMMS), China (Assurance Number: Consent: Convalescent sera were collected from COVID-19 patients from Beijing YouAn Hospital, Capital Medical University with written informed consent.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   ELISA: ELISA was performed to detect SARS-CoV-2 RBD and S1-specific IgG antibodies in the immunized mouse or Macaca fascicularis …

   SciScore for 10.1101/2020.11.29.402339: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	IACUC: All animal procedures were reviewed and approved by the Animal Experiment Committee of Laboratory Animal Center, Academy of Military Medical Sciences (AMMS), China (Assurance Number: Consent: Convalescent sera were collected from COVID-19 patients from Beijing YouAn Hospital, Capital Medical University with written informed consent.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   ELISA: ELISA was performed to detect SARS-CoV-2 RBD and S1-specific IgG antibodies in the immunized mouse or Macaca fascicularis sera.	S1-specific IgG suggested: None
   After four washes, the bound antibodies were detected by incubation with horseradish peroxidase (HRP)-conjugated anti-mouse IgG or anti-Macaca fascicularis IgG antibody (1:5,000, Thermo Fisher Scientific) for 30 min at 37°C.	anti-mouse IgG suggested: None anti-Macaca fascicularis IgG suggested: None
   Medium and anti-CD3, anti-CD28 antibody were used for negative and positive controls, respectively.	anti-CD3 suggested: None anti-CD28 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   For protein expression, the plasmid was first transfected into CHO-K1 cells and then stable clones were isolated by the limiting dilution method in the presence of 50 μM of Methionine Sulfoximine (MSX).	CHO-K1 suggested: None
   Patient-derived SARS-CoV-2 isolates including 1) BetaCoV/Beijing/IMEBJ01/2020 (Patients originated from Beijing China; Accession No: GWHACAX01000000; Abbreviated as BJ01); 2) BetaCoV/Beijing/IMEBJ05/2020(Patients originated from Wuhan, China; Accession Nos: GWHACBB01000000. Abbreviated as BJ05);3) BetaCoV/Beijing/IMEBJ08/2020 (Patients originated from: Italy, Accession Nos: GWHAMKA01000000, Abbreviated as BJ08) were passaged in Vero cells and the virus stock was aliquoted and titrated to PFU/ml in Vero cells by plaque assay22.	Vero suggested: None
   In brief, mice or Macaca fascicularis sera at 3-fold serial dilutions were incubated with 650 TCID50 of the pseudovirus for 1 hour at 37 °C, and then 20000 Huh7 cells were added into each well.	Huh7 suggested: CLS Cat# 300156/p7178_HuH7, RRID:CVCL_0336)
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Animal immunization and challenge studies: Eight-week old human ACE2 transgenic mice (hACE2-Tg, obtained from National Institutes for Food and Drug Control, Beijing, China), Balb/C mice and 3-year old Macaca fascicularis (all from Laboratory animal center of the academic of the military medical sciences) were immunized with SARS-CoV-2 RBD-Fc Vacc as described in Fig.1a.	Balb/C suggested: None
   intraocular 0.2ml; hACE2-Tg mice were inoculated intranasally with 40 μL of BetaCoV(BJ05).	hACE2-Tg suggested: None
   Software and Algorithms
   Sentences	Resources
   Cell events were acquired using an FACS CANTO (BD), followed by FlowJo software (FlowJo LLC, Ashland, OR) analysis.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   Statistical analysis: Statistical analyses were carried out using Prism software (GraphPad).	Prism suggested: (PRISM, RRID:SCR_005375) GraphPad 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 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.
   • 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 10.1101/2020.11.29.402339v1 on bioRxiv

   Nov 30, 2020