Induction of neutralizing antibodies against SARS-CoV-2 variants by a multivalent mRNA-lipid nanoparticle vaccine encoding SARS-CoV-2/SARS-CoV Spike protein receptor-binding domains

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   ScreenIT

   Apr 30, 2022

   SciScore for 10.1101/2022.04.28.489834: (What is this?)

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

   Table 1: Rigor

   Ethics	not detected.
   Sex as a biological variable	Immunization of mice with RBD mRNA-LNPs: Male C57BL/6J mice (aged 4–5 weeks) were purchased from the Jackson Laboratory and housed according to the regulatory standards of the University of California, San Diego.
   Randomization	Mice were randomly allocated to experimental groups.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	Contamination: The cell lines were tested and confirmed to be negative for mycoplasma.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   Cell lines: HEK293FT and Vero E6 cells were maintained in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum (GIBCO).	HEK293FT suggested: None

   SciScore for 10.1101/2022.04.28.489834: (What is this?)

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

   Table 1: Rigor

   Ethics	not detected.
   Sex as a biological variable	Immunization of mice with RBD mRNA-LNPs: Male C57BL/6J mice (aged 4–5 weeks) were purchased from the Jackson Laboratory and housed according to the regulatory standards of the University of California, San Diego.
   Randomization	Mice were randomly allocated to experimental groups.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	Contamination: The cell lines were tested and confirmed to be negative for mycoplasma.

   Table 2: Resources

   Experimental Models: Cell Lines
   Sentences	Resources
   Cell lines: HEK293FT and Vero E6 cells were maintained in Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum (GIBCO).	HEK293FT suggested: None Vero E6 suggested: None
   SARS-CoV-2 pseudovirus production: Plasmids encoding the Spike proteins (lacking the C-terminal 19-amino acids) of SARS-CoV-1 (CUHK-W1), SARS-CoV-2 (Wuhan-Hu-1), variant B.1.351, variant B.1.617.2, Wuhan-N501Y, and Wuhan-E484K were transfected into 293T cells with Lipofectamine 3000 (ThermoFisher).	293T suggested: None
   Verification of protein-coding capability of vaccine mRNAs: To confirm that the synthesized mRNAs could be translated into GFP or RBD proteins, 293FT cells were seeded at 3 × 105 cells/mL in 6-well plates, grown for 24 h, and then transfected with 1 mg mRNA per well using Lipofectamine 3000 (Invitrogen) according to the manufacturer’s instructions.	293FT suggested: None
   Vero cells were seeded at 2 × 105 cells/mL of 100 μL/well in 96-well plates and cultured overnight at 37°C.	Vero suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Immunization of mice with RBD mRNA-LNPs: Male C57BL/6J mice (aged 4–5 weeks) were purchased from the Jackson Laboratory and housed according to the regulatory standards of the University of California, San Diego.	C57BL/6J suggested: RRID:IMSR_JAX:000664)
   Software and Algorithms
   Sentences	Resources
   Statistical analyses were conducted using GraphPad Prism 8.0.	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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   Results from scite Reference Check: We found no unreliable references.

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   Version published to 10.1101/2022.04.28.489834v1 on bioRxiv

   Apr 28, 2022