Polymersomes Decorated with the SARS-CoV-2 Spike Protein Receptor-Binding Domain Elicit Robust Humoral and Cellular Immunity
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SciScore for 10.1101/2021.04.08.438884: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: Mouse vaccination experiments: All experiments were performed in accordance with the Institutional Animal Care and Use Committee at the University of Chicago. Randomization Female 8-week-old C57BL/6 mice (Jackson Laboratory) were randomly assigned to cohorts of n = 5 and vaccinated with 10 μg of antigen and 5 μg of adjuvant s. Blinding not detected. Power Analysis not detected. Sex as a biological variable Female 8-week-old C57BL/6 mice (Jackson Laboratory) were randomly assigned to cohorts of n = 5 and vaccinated with 10 μg of antigen and 5 μg of adjuvant s. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Wells were then washed … SciScore for 10.1101/2021.04.08.438884: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement IACUC: Mouse vaccination experiments: All experiments were performed in accordance with the Institutional Animal Care and Use Committee at the University of Chicago. Randomization Female 8-week-old C57BL/6 mice (Jackson Laboratory) were randomly assigned to cohorts of n = 5 and vaccinated with 10 μg of antigen and 5 μg of adjuvant s. Blinding not detected. Power Analysis not detected. Sex as a biological variable Female 8-week-old C57BL/6 mice (Jackson Laboratory) were randomly assigned to cohorts of n = 5 and vaccinated with 10 μg of antigen and 5 μg of adjuvant s. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Wells were then washed five times with PBS-T and incubated for an additional hour at RT with horseradish peroxide (HRP)-conjugated antibodies against mouse IgG, IgG1, IgG2b, IgG3, or IgA (Southern Biotech). mouse IgGsuggested: NoneIgG1suggested: NoneIgG2bsuggested: NoneIgG3suggested: NoneIgA ( Southern Biotech)suggested: NoneExperimental Models: Cell Lines Sentences Resources Suspension-adapted HEK-293F cells were maintained in serum-free Free Style 293 Expression Medium (Gibco). HEK-293Fsuggested: RRID:CVCL_6642)MPLA PS in vitro activity: To determine TLR4 activation, HEK-Blue™ TLR4 cells (Invivogen) were incubated with increasing concentrations of MPLA PS for 24 h 37 °C in a 5% CO2 incubator. HEK-Blue™ TLR4suggested: NoneRBDsurfin vitro activity: For the cell-based hACE2-binding assay, human embryonic kidney (HEK)-293T cells overexpressing human ACE-2 (HEK-hACE2) were obtained from BEI Resources (NIH NIAID) HEK)-293Tsuggested: NoneFluorescently labeled empty PS, RBDsurf, or RBDfree was incubated at varying concentrations with 5 × 104 HEK-hACE2 cells at 4 °C for 20 min. HEK-hACE2suggested: NoneThe pre-incubated viruses were then applied to Vero-E6 cell monolayers, and the cells were maintained until > 90% cell death for the negative control (4-5 d). Vero-E6suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Female 8-week-old C57BL/6 mice (Jackson Laboratory) were randomly assigned to cohorts of n = 5 and vaccinated with 10 μg of antigen and 5 μg of adjuvant s. C57BL/6suggested: NoneSoftware and Algorithms Sentences Resources Gels were imaged on a ChemiDoc XRS+ Gel Documentation System (Bio-Rad) and analyzed using ImageJ. ImageJsuggested: (ImageJ, RRID:SCR_003070)RBDsurfin vitro activity: For the cell-based hACE2-binding assay, human embryonic kidney (HEK)-293T cells overexpressing human ACE-2 (HEK-hACE2) were obtained from BEI Resources (NIH NIAID) NIAIDsuggested: (NIAID, RRID:SCR_016598)Samples were acquired on a BD LSR-Fortessa (BD) and analyzed using FlowJo™ software. FlowJo™suggested: (FlowJo, RRID:SCR_008520)Cells were subsequently fixed and permeabilized using BD Cytofix/Cytoperm™, and intracellular cytokines were stained using fluorescent-mAbs listed in Supplementary Table 3. BD Cytofix/Cytoperm™suggested: NonePeptide array analysis: Antibody specificity to linear epitopes of the spike protein was analyzed using a CelluSpots™ Covid19_hullB Peptide Array ( Antibodysuggested: (Antibodypedia, RRID:SCR_012782)Spots were analyzed using Spotfinder software (version v3.2.1) Spotfindersuggested: (Spotfinder, RRID:SCR_000085)Software packages and statistical analysis: Statistical analysis was performed using GraphPad Prism 9 (GraphPad). GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)All flow cytometry data were analyzed using FlowJo_v10.7.2 software (FlowJo LLC, BD Biosciences). FlowJosuggested: (FlowJo, RRID:SCR_008520)Figures 1 and 2 were created using BioRender (https://biorender.com) as part of an Academic License through the Chicago Immunoengineering Innovation Center. BioRendersuggested: (Biorender, RRID:SCR_018361)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: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.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.
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