An mRNA vaccine against SARS-CoV-2: Lyophilized, liposome-based vaccine candidate EG-COVID induces high levels of virus neutralizing antibodies

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1. 

   ScreenIT

   Mar 26, 2021

   SciScore for 10.1101/2021.03.22.436375: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	Immunization of mice: For immunogenicity studies, 6-week-old female B6C3F1 mice (SLC, Japan) were randomly assigned into experimental groups (n = 6).
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Bioluminescence imaging: For bioluminescence imaging studies, 6-week-old female Balb/c nude or C57BL/6N mice (SLC, Japan) were used.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The primary and secondary antibodies were SARS-CoV-2-S1 (Novus, Centennial, CO, USA), beta-actin (Cell signaling, Danvers, MA, USA) and anti-rabbit (Invitrogen, Thermo Scientific, Waltham, MA, USA).	beta-ac…

   SciScore for 10.1101/2021.03.22.436375: (What is this?)

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

   Table 1: Rigor

   Institutional Review Board Statement	not detected.
   Randomization	Immunization of mice: For immunogenicity studies, 6-week-old female B6C3F1 mice (SLC, Japan) were randomly assigned into experimental groups (n = 6).
   Blinding	not detected.
   Power Analysis	not detected.
   Sex as a biological variable	Bioluminescence imaging: For bioluminescence imaging studies, 6-week-old female Balb/c nude or C57BL/6N mice (SLC, Japan) were used.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   The primary and secondary antibodies were SARS-CoV-2-S1 (Novus, Centennial, CO, USA), beta-actin (Cell signaling, Danvers, MA, USA) and anti-rabbit (Invitrogen, Thermo Scientific, Waltham, MA, USA).	beta-actin suggested: None anti-rabbit suggested: None
   Antigen-specific antibody titer was determined by end-point dilution ELISA.	Antigen-specific suggested: None
   Bound antibodies were detected with HRP-conjugated goat anti-mouse total IgG antibody (Jackson, PA, USA) diluted 1:5000 for 1 hour at 37°C.	anti-mouse total IgG suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Cell culture and mRNA transfection: HEK-293T cell was obtained from ATCC (Manasass, VA, USA).	HEK-293T suggested: CCLV Cat# CCLV-RIE 1018, RRID:CVCL_0063)
   EG-COVID 001, 002, 003 and 004 (10 ug mRNA containg complex each) were introduced into 293T cells by 18 hour incubation in OPTi-MEM (Thermo Scientific, MA, USA).	293T suggested: None
   Vero cells were seeded at 8 × 105 cells/well in 6 well plates and cultured overnight.	Vero suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Bioluminescence imaging: For bioluminescence imaging studies, 6-week-old female Balb/c nude or C57BL/6N mice (SLC, Japan) were used.	Balb/c nude suggested: None C57BL/6N suggested: RRID:MGI:5825016)
   Immunization of mice: For immunogenicity studies, 6-week-old female B6C3F1 mice (SLC, Japan) were randomly assigned into experimental groups (n = 6).	B6C3F1 suggested: RRID:IMSR_JAX:100010)

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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 28. 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.
   • 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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2. 

   Version published to 10.1101/2021.03.22.436375v2 on bioRxiv

   Mar 24, 2021
3. 

   Version published to 10.1101/2021.03.22.436375v1 on bioRxiv

   Mar 22, 2021