Nanoparticle-delivered TLR4 and RIG-I agonists enhance immune response to SARS-CoV-2 subunit vaccine

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1. Version published to 10.1016/j.jconrel.2022.05.023

   Jul 1, 2022
2. 

   ScreenIT

   Feb 5, 2022

   SciScore for 10.1101/2022.01.31.478507: (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	34 Female Balb/cJ (5-10 weeks old, Jackson Labs) were euthanized with CO2.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	The second emulsion was sonicated at 65% power for 5 min at RT.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   A secondary biotinylated anti-mouse IgA, total IgG, IgG1, or IgG2a antibody (SouthernBiotech) was diluted 5,000-fold in 5-fold diluted PBSTBA and was added to plates for 2 h at RT.	A secondary biotinylated anti-mouse IgA, total IgG, IgG1, or IgG2a antibody (SouthernBiotech) suggested: None anti-mouse IgA, total IgG suggested: None IgG1 suggested: None IgG2a suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Re…

   SciScore for 10.1101/2022.01.31.478507: (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	34 Female Balb/cJ (5-10 weeks old, Jackson Labs) were euthanized with CO2.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	The second emulsion was sonicated at 65% power for 5 min at RT.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   A secondary biotinylated anti-mouse IgA, total IgG, IgG1, or IgG2a antibody (SouthernBiotech) was diluted 5,000-fold in 5-fold diluted PBSTBA and was added to plates for 2 h at RT.	A secondary biotinylated anti-mouse IgA, total IgG, IgG1, or IgG2a antibody (SouthernBiotech) suggested: None anti-mouse IgA, total IgG suggested: None IgG1 suggested: None IgG2a suggested: None
   Experimental Models: Organisms/Strains
   Sentences	Resources
   To isolate T cells, spleens from female Balb/cJ mice were dissociated in 2 mg/mL Collagenase D (Sigma-Aldrich, Cat# 11088882001) in Opti-MEM media (Gibco, Cat# 11058021) and filtered through a 40 μm cell strainer.	Balb/cJ suggested: None
   Female BALB/c mice were intramuscularly injected with adjuvant-loaded NPs (4 mg/mouse) and variable doses of stabilized SARS-CoV-2 spike glycoprotein (BEI Resources, Cat# NR-52397) into the left and right anterior tibialis anterior muscles (50 μL NPs in saline per injection) on day 0 (prime) and day 28 (boost).	BALB/c suggested: None
   Software and Algorithms
   Sentences	Resources
   MPLA loading estimation via GC-MS, LC-MS, and surrogate fluorometry has been previously described.28 PUUC and CpG loading was quantified by supernatant measurement of unbound RNA or DNA, respectively, with the Nucleic Acid Quantification workflow on a Synergy HT plate reader (BioTek) with Gen5 software.	Gen5 suggested: (Gen5, RRID:SCR_017317)
   FlowJo Software (BD) was used to generate t-distributed stochastic neighbor embedding (t-SNE) plots.	FlowJo suggested: (FlowJo, RRID:SCR_008520)
   Statistical analyses were performed with GraphPad Prism 9.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)
   For antibody quantification with ELISA, area under the curve (AUC) across fold dilutions was computed with the AUC function on GraphPad as previously reported.	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 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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3. 

   Version published to 10.1101/2022.01.31.478507v1 on bioRxiv

   Feb 2, 2022