Lyophilized mRNA-lipid nanoparticle vaccines with long-term stability and high antigenicity against SARS-CoV-2

Article activity feed

1. Version published to 10.1038/s41421-022-00517-9

   Jan 23, 2023
2. 

   Version published to 10.1101/2022.02.10.479867v3 on bioRxiv

   Jul 25, 2022
3. 

   ScreenIT

   Feb 15, 2022

   SciScore for 10.1101/2022.02.10.479867: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Animal experiments were executed by certified staff in Center for Animal Experiments of Wuhan University, approved by the Institutional Animal Care and Use Committee (AUP #WP2021-0607).
   Sex as a biological variable	2.5 Cell culture: HEK 293T/17 and ACE2-expressing 293T cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 2 mM L-glutamine, 10% FBS (sigma-aldrich) and 1% penicillin/streptomycin (BI) at 37 °C and 5% CO2. 2.6 Animals: Female BALB/c mice age of 6-8 weeks and heterozygous B6/JGpt-H11em1Cin(K18-ACE2)/Gpt mice (K18-hACE2 KI mice) age of 5-7 weeks and body weight of 18-20 g were used.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not …

   SciScore for 10.1101/2022.02.10.479867: (What is this?)

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

   Table 1: Rigor

   Ethics	IACUC: Animal experiments were executed by certified staff in Center for Animal Experiments of Wuhan University, approved by the Institutional Animal Care and Use Committee (AUP #WP2021-0607).
   Sex as a biological variable	2.5 Cell culture: HEK 293T/17 and ACE2-expressing 293T cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 2 mM L-glutamine, 10% FBS (sigma-aldrich) and 1% penicillin/streptomycin (BI) at 37 °C and 5% CO2. 2.6 Animals: Female BALB/c mice age of 6-8 weeks and heterozygous B6/JGpt-H11em1Cin(K18-ACE2)/Gpt mice (K18-hACE2 KI mice) age of 5-7 weeks and body weight of 18-20 g were used.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Following washes, the secondary antibody, goat Anti-Mouse IgG H&L conjugated HRP (Abcam), was incubated in plate at room temperature for 1 h.	goat Anti-Mouse IgG H&L conjugated HRP suggested: None Anti-Mouse IgG suggested: None
   2.11 SARS-CoV-2 Delta virus neutralization assay: SARS-CoV-2 Delta-neutralizing antibody was determined by in vitro inhibition of cytopathic effect (CPE), and was performed in the ABSL-3 lab at Center of Laboratory Animal Sciences, Wuhan University (Wuhan, China).	CPE suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   2.5 Cell culture: HEK 293T/17 and ACE2-expressing 293T cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 2 mM L-glutamine, 10% FBS (sigma-aldrich) and 1% penicillin/streptomycin (BI) at 37 °C and 5% CO2. 2.6 Animals: Female BALB/c mice age of 6-8 weeks and heterozygous B6/JGpt-H11em1Cin(K18-ACE2)/Gpt mice (K18-hACE2 KI mice) age of 5-7 weeks and body weight of 18-20 g were used.	HEK 293T/17 suggested: ATCC Cat# CRL-11268, RRID:CVCL_1926) 293T suggested: None
   After that, the 293T-ACE2 cells are added and incubated for 24 h.	293T-ACE2 suggested: None
   The virus-serum mixtures were added to Vero-E6 cells seeded in 24-well plate and cultured in a 5% CO2 incubator at 37 °C for 3 days.	Vero-E6 suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   2.5 Cell culture: HEK 293T/17 and ACE2-expressing 293T cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 2 mM L-glutamine, 10% FBS (sigma-aldrich) and 1% penicillin/streptomycin (BI) at 37 °C and 5% CO2. 2.6 Animals: Female BALB/c mice age of 6-8 weeks and heterozygous B6/JGpt-H11em1Cin(K18-ACE2)/Gpt mice (K18-hACE2 KI mice) age of 5-7 weeks and body weight of 18-20 g were used.	BALB/c suggested: None B6/JGpt-H11em1Cin(K18-ACE2)/Gpt suggested: None K18-hACE2 KI suggested: None
   Software and Algorithms
   Sentences	Resources
   The mRNA integrity was measured with a gel retardation assay [15] and microfluidic capillary electrophoresis (Agilent Fragment Analyser) [16], as previously reported.	Agilent Fragment Analyser suggested: (OMICtools, RRID:SCR_002250)
   Statistical analysis was mainly processed using Prism software version 8.3	Prism suggested: (PRISM, RRID:SCR_005375)
   GraphPad Software Inc.	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: Please consider improving the rainbow (“jet”) colormap(s) used on page 9. 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.
   • No funding statement was detected.
   • 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
4. 

   Version published to 10.1101/2022.02.10.479867v2 on bioRxiv

   Feb 11, 2022
5. 

   Version published to 10.1101/2022.02.10.479867v1 on bioRxiv

   Feb 10, 2022