SARS-CoV-2 mRNA-vaccine candidate; COReNAPCIN®, induces robust humoral and cellular immunity in mice and non-human primates

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1. Version published to 10.1038/s41541-022-00528-3

   Sep 2, 2022
2. 

   ScreenIT

   Mar 10, 2022

   SciScore for 10.1101/2022.03.05.483092: (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	Total number of 80 BALB/c mice in four groups (15 females and 5 males per group), and 48 C57BL/6 mice in four groups (12 females in each group), were injected intramuscularly (IM) with either phosphate-buffered saline (PBS) or 0.05, 0.5, or 3 μg mRNA-LNP at days 0 and 21.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After 2 hrs blocking in a buffer containing 5% fat-free dried milk and 0.5% Tween-20, the membrane was incubated with anti-SARS-CoV-2 spike polyclonal antibody (Sino biological) or anti-β-actin polyclonal antibody (Abcam) …

   SciScore for 10.1101/2022.03.05.483092: (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	Total number of 80 BALB/c mice in four groups (15 females and 5 males per group), and 48 C57BL/6 mice in four groups (12 females in each group), were injected intramuscularly (IM) with either phosphate-buffered saline (PBS) or 0.05, 0.5, or 3 μg mRNA-LNP at days 0 and 21.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   After 2 hrs blocking in a buffer containing 5% fat-free dried milk and 0.5% Tween-20, the membrane was incubated with anti-SARS-CoV-2 spike polyclonal antibody (Sino biological) or anti-β-actin polyclonal antibody (Abcam) overnight at 4°C.	anti-SARS-CoV-2 suggested: None anti-β-actin suggested: None
   Flow cytometry: In order to analyze transfected cells for cell surface expression of S protein, 2×105 cells of each of transfected and untransfected cells stained with SARS-CoV-2 Spike antibody (SinoBiological) and Anti-human IgG FITC (Sigma-Aldrich®).	Anti-human IgG suggested: None
   After incubation with detection antibody (7-B6-ALP) and subsequent washing, 100 μl substrate solution (BCIP/NBT-plus) added to each well.	7-B6-ALP suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   In vitro characterization of mRNA-LNP: 4-4-1 Cell transfection: Primarily, HEK 293T cells were seeded into a 12-well cell culture plate, 2×105 cells per well in HG-DMEM medium (Biowest).	HEK 293T suggested: None
   After 60 min incubation at 37°C, the virus-serum mixture was added to 96 well-plate previously seeded with 1.5 × 104 Vero cells per well (performed in triplicate).	Vero suggested: CLS Cat# 605372/p622_VERO, RRID:CVCL_0059)
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Animal models and immunization: Immunogenicity of COReNAPCIN® vaccine candidate was assessed in mice (BALB/c and C57BL/6) and rhesus macaques.	BALB/c suggested: None
   Mice immunization: BALB/c and C57BL/6 mice (16-18 g, 4-6 weeks old) were purchased from the Pasteur Institute of Iran.	C57BL/6 suggested: None
   Recombinant DNA
   Sentences	Resources
   This DNA sequence was cloned into ReNAP-IVT1 plasmid in between special 5’-UTR and 3’-UTR-polyA tail, for transcribing the mRNA under the control of a T7 promoter.	ReNAP-IVT1 suggested: None
   Spike pseudotyped lentivirus, produced by the co-transfection of plex307-egfp, pCMV3-spike (Wuhan-D614G), and pspax2 with Lipofectamine 3000 (Thermo)	pCMV3-spike suggested: None
   Software and Algorithms
   Sentences	Resources
   IC50 was calculated using the percentage of GFP positive cell versus log (dilution factor) in GraphPad Prism V8. 4.9	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   After 20 minutes incubation and washing with permeabilization and staining buffer (Biolegend), cells were acquired on BD FACS Lyrics and analyzed by Flowjo V10 (BD Biosciences). 4.12.	Flowjo suggested: (FlowJo, RRID:SCR_008520)
   Statistical analysis: Graph Pad V8 Prism was used to illustrates the figures and analyze the results statistically.	Prism suggested: (PRISM, RRID:SCR_005375)

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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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   Read the original source
3. 

   Version published to 10.1101/2022.03.05.483092v1 on bioRxiv

   Mar 7, 2022