Comparison of Wild Type DNA Sequence of Spike Protein from SARS-CoV-2 with Optimized Sequence on The Induction of Protective Responses Against SARS-Cov-2 Challenge in Mouse Model

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1. Version published to 10.1080/21645515.2021.2016201

   Jan 21, 2022
2. 

   ScreenIT

   Aug 19, 2021

   SciScore for 10.1101/2021.08.13.456164: (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	Animal experiment: Female, C57BL/6 mice (6-8 weeks of age) and Balb/c mice (6-8 weeks of age) were purchased from Beijing Vital Laboratory Animal Technology Co., Ltd. (Beijing, China) and Shanghai Jiesjie Laboratory Animal Co., Ltd. (Shanghai, China), which were kept in SPF condition.
   Randomization	All animal studies were performed after randomization.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Antigen-specific humoral immune responses: Enzyme-linked immunosorbent assay (ELISA) was used to measure the antigen-specific antibody production induced by the DNA …

   SciScore for 10.1101/2021.08.13.456164: (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	Animal experiment: Female, C57BL/6 mice (6-8 weeks of age) and Balb/c mice (6-8 weeks of age) were purchased from Beijing Vital Laboratory Animal Technology Co., Ltd. (Beijing, China) and Shanghai Jiesjie Laboratory Animal Co., Ltd. (Shanghai, China), which were kept in SPF condition.
   Randomization	All animal studies were performed after randomization.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Antigen-specific humoral immune responses: Enzyme-linked immunosorbent assay (ELISA) was used to measure the antigen-specific antibody production induced by the DNA vaccinations as previously described.	antigen-specific suggested: None
   Antibodies were detected with HRP-conjugated goat anti-mouse IgG (Southern Biotech, Birmingham, AL) after the enzymatic reaction was developed, the OD values were read at 450/620 nm by an ELISA plate reader (Bio-Rad, Hercules, CA).	anti-mouse IgG suggested: (Jackson ImmunoResearch Labs Cat# 015-620-003, RRID:AB_2337231)
   The Goat anti-Rabbit IgG-HRP (Invitrogen, 32460) and Rabbit anti-hACE2 antibody (Sino Biological, 10108-RP01) were used to detect the concentration of hACE2 binding with the RBD.	anti-Rabbit IgG-HRP suggested: None anti-hACE2 suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   Seed SARS-CoV-2 (SARS-CoV-2/WH-09/human/2020) stocks and virus isolation studies were performed in Vero E6 cells, which are maintained in Dulbecco’s modified Eagle’s medium (DMEM, Invitrogen, USA) supplemented with 10% fetal bovine serum (FBS), 100 IU/mL penicillin, and 100 μg /mL streptomycin, and incubated at 36.5 °C, 5% CO2.	Vero E6 suggested: RRID:CVCL_XD71)
   The diluted samples were mixed with a virus suspension of 100 TCID50 in 96-well plates at a ratio of 1:1, followed by 2 h incubation at 36.5 °C in a 5% CO2 incubator. 1× 104 Vero cells were then added to the serum-virus mixture, and the plates were incubated for 3–5 days at 36.5 °C in a 5% CO2 incubator.	Vero suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   Animal experiment: Female, C57BL/6 mice (6-8 weeks of age) and Balb/c mice (6-8 weeks of age) were purchased from Beijing Vital Laboratory Animal Technology Co., Ltd. (Beijing, China) and Shanghai Jiesjie Laboratory Animal Co., Ltd. (Shanghai, China), which were kept in SPF condition.	C57BL/6 suggested: None Balb/c suggested: None
   Recombinant DNA
   Sentences	Resources
   Plasmid preparation: Plasmids of pGX9501, pVAX1-S-WT, and pVAX1 were transformed into DH5a E.	pGX9501 suggested: None pVAX1-S-WT suggested: None
   Software and Algorithms
   Sentences	Resources
   Six hours later, the percentage of labeled cells was detected with LSRFortessa flow cytometry (BD) and analyzed by FlowJo (TreeStar).	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   Statistics were performed using GraphPad Prism 7 software.	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 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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   Results from scite Reference Check: We found no unreliable references.

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   Read the original source
3. 

   Version published to 10.1101/2021.08.13.456164v1 on bioRxiv

   Aug 16, 2021