Next-generation intranasal Covid-19 vaccine: a polymersome-based protein subunit formulation that provides robust protection against multiple variants of concern and early reduction in viral load of the upper airway in the golden Syrian hamster model

Article activity feed

1. 

   ScreenIT

   Feb 16, 2022

   SciScore for 10.1101/2022.02.12.480188: (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	Golden Syrian hamsters comprising equal numbers of males and females were purchased from an approved vendor.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   …

   Antibodies
   Sentences	Resources
   Serum IgG ELISA: Indirect ELISA was performed to analyze sera for binding antibodies to SARS-CoV-2 spike protein.	SARS-CoV-2 spike protein . suggested: None
   Secondary detection antibody (goat anti-species-HRP IgG; Abcam) was added at a dilution of 1:10,000 in 100 µl per well.	goat anti-species-HRP IgG; Abcam suggested: None anti-species-HRP IgG suggested: None
   Experimental Models: Cell Lines
   Sentences

   SciScore for 10.1101/2022.02.12.480188: (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	Golden Syrian hamsters comprising equal numbers of males and females were purchased from an approved vendor.
   Randomization	not detected.
   Blinding	not detected.
   Power Analysis	not detected.
   Cell Line Authentication	not detected.

   Table 2: Resources

   Antibodies
   Sentences	Resources
   Serum IgG ELISA: Indirect ELISA was performed to analyze sera for binding antibodies to SARS-CoV-2 spike protein.	SARS-CoV-2 spike protein . suggested: None
   Secondary detection antibody (goat anti-species-HRP IgG; Abcam) was added at a dilution of 1:10,000 in 100 µl per well.	goat anti-species-HRP IgG; Abcam suggested: None anti-species-HRP IgG suggested: None
   Experimental Models: Cell Lines
   Sentences	Resources
   In brief, 2.4 x105 Sf9 insect cells were plated into a well of a 24 well plate.	Sf9 suggested: None
   Plaque reduction neutralization test (PRNT): Vero 76 cells were cultured in 24-well plates at 175,000 cells/well in DMEM + 10% v/v FBS + Gentamicin and incubated at 37 °C, 5% CO2.	Vero 76 suggested: None
   Subsequently, Vero cell culture medium was removed from 24-well plate and 250 μl of titrated serum samples were added in duplicates.	Vero suggested: None
   Recombinant DNA
   Sentences	Resources
   Baculovirus production: The Beta variant spike protein ectodomain gene (amino acids 1-1201) containing the native signal peptide, 3Q mutations to the furin cleavage site and 2P mutations, was codon-optimized for insect cell protein expression, using GenScript proprietary algorithm and was directly synthesized into pFAST-BAC1 transfer plasmid.	pFAST-BAC1 suggested: None
   To generate a control for the amplification reaction, RNA was isolated from the applicable SARS-CoV-2 plasmid control using the same procedure. qPCR was set up using TaqMan Fast Virus 1-Step Real-time RT-PCR protocol with assay setup performed using Qiagen Qiagility automated PCR setup platform and analyzed in Applied Biosystems on QuantStudio 3.	SARS-CoV-2 suggested: RRID:Addgene_164583)
   Software and Algorithms
   Sentences	Resources
   Encapsulation of antigen were quantified by densiometric analysis using a known S1S2 protein standards in Fiji ImageJ software (v.	ImageJ suggested: (ImageJ, RRID:SCR_003070)
   Serum titration curves were analyzed by 5-parameter, non-linear regression using GraphPad Prism version 8.4.3 to determine antibody titers.	GraphPad Prism suggested: (GraphPad Prism, RRID:SCR_002798)

   ---

   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).

   ---

   Results from LimitationRecognizer: We detected the following sentences addressing limitations in the study:

   The main limitation of the present study was that T cell responses had not been investigated, as commercialized antibodies required for assessment of hamster T cell subsets and function were largely unavailable 66. Concerns over such constraints of the hamster model had been raised 67. Nevertheless, our previous mouse study did establish the presence of functional, memory CD4+ and CD8+ T cells after primary vaccination 34. Unlike neutralizing antibodies, which generally exhibited narrow target specificity as evidenced by moderate to severe reduction in potency against different variants, T cell epitopes were remarkably conserved 24. Studies had shown CD4+ and CD8+ T cells from previous vaccination or infection to retain robust activity against Omicron, despite the variant’s extensive mutations and increased resistance to neutralizing antibodies 22, 23. It was believed that cross-reactive T cells may contribute to the control Omicron infection and possibly account for the reduction in disease severity compared to the earlier Delta wave 68.

   ---

   Results from TrialIdentifier: No clinical trial numbers were referenced.

   ---

   Results from Barzooka: We did not find any issues relating to the usage of bar graphs.

   ---

   Results from JetFighter: We did not find any issues relating to colormaps.

   ---

   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.

   ---

   Results from scite Reference Check: We found no unreliable references.

   ---

   Read the original source
2. 

   Version published to 10.1101/2022.02.12.480188v1 on bioRxiv

   Feb 14, 2022