An mRNA SARS-CoV-2 Vaccine Employing Charge-Altering Releasable Transporters with a TLR-9 Agonist Induces Neutralizing Antibodies and T Cell Memory
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SciScore for 10.1101/2021.04.14.439891: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Serum preparation: For human samples, informed consent was obtained from the subjects prior to blood draw. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Mouse vaccination: Eight-to twelve-week-old female BALB/c mice were purchased from The Jackson Laboratory and housed in the Laboratory Animal Facility of the Stanford University Medical Center. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources In some assays, a human anti-RBD (Invivogene) of known antibody concentration was used as standard. anti-RBDsuggested: NoneRBD was then detected using an Alexa Fluor 488 … SciScore for 10.1101/2021.04.14.439891: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement Consent: Serum preparation: For human samples, informed consent was obtained from the subjects prior to blood draw. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable Mouse vaccination: Eight-to twelve-week-old female BALB/c mice were purchased from The Jackson Laboratory and housed in the Laboratory Animal Facility of the Stanford University Medical Center. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources In some assays, a human anti-RBD (Invivogene) of known antibody concentration was used as standard. anti-RBDsuggested: NoneRBD was then detected using an Alexa Fluor 488 conjugated anti-His antibody (clone J099B1, Biolegend). anti-Hissuggested: NoneAs positive control a neutralizing human anti-SARS-Cov-2 IgG1 antibody was used (Acro). anti-SARS-Cov-2 IgG1suggested: NoneCells were cultured in 96-well plates (Corning, V-bottom) at 1×106 cells/well and stimulated for 48 hours with 5 μg/mL of RBD-His or hCD81-His or media alone (RPMI + 5% FCS) in the presence of anti-mouse CD28 antibody [0,5 µg/ml] (Southern Biotech). hCD81-Hissuggested: Noneanti-mouse CD28suggested: NoneFollowing stimulation, cells were washed, stained with Aqua Live/dead viability dye (Thermo Fisher) in PBS, washed two additional times and stained with a cocktail of monoclonal antibodies and Fc block: CD16/32, CD4 BV605 RM4-5, CD8 FITC 53-6.7, CD44 APC IM7, CD69 PE-Cy7 H1.2F3, CD134 BV786 OX-86 CD44suggested: (BD Biosciences Cat# 563736, RRID:AB_2738395)CD69suggested: (BD Biosciences Cat# 563834, RRID:AB_2738441)CD134suggested: (BD Biosciences Cat# 744746, RRID:AB_2742454)As positive control cells were stimulated with anti-mouse CD3 [0,05 µg/ml] (Southern Biotech) and anti-mouse CD28 antibody [0,5 µg/ml] (Southern Biotech). anti-mouse CD3suggested: NoneFollowing stimulation, cells were washed, stained with Aqua Live/dead viability dye (Thermo Fisher) in PBS, washed two additional times and stained with a cocktail of monoclonal antibodies and Fc block: CD16/32, CD4 Ax700 RM4-5, CD8 APC-H7 53-6.7, CD4suggested: (Miltenyi Biotec Cat# 130-109-536, RRID:AB_2657974)Tests were performed in triplicates and with a positive control (anti-CD3 monoclonal antibody (0,05 µg/ml; Southern Biotech) anti-CD3suggested: NoneAfter stimulation wells were washed and incubated with a biotinylated anti-IFNγ antibody (R&D systems, mouse IFNγ Kit Cat # EL485) overnight at 4°C. anti-IFNγsuggested: NoneExperimental Models: Cell Lines Sentences Resources Hela and 293F transfection: Hela cells and 293F cells were plated at 106 cells per well in a 12 well plate in Opti-MEM media (ThermoFisher Scientific). 293Fsuggested: NoneOne day before transfection, 6 x 106 HEK293T cells were seeded in a 10-cm culture plate in RPMI containing 10% FCS, 2mM L-glutamine, streptomycin and penicillin. HEK293Tsuggested: NoneFor viral neutralization assays, ACE2-expressing HEK293T46 cells were plated in poly-L-lysine-coated, white-walled HEK293T46suggested: NoneExperimental Models: Organisms/Strains Sentences Resources Mouse vaccination: Eight-to twelve-week-old female BALB/c mice were purchased from The Jackson Laboratory and housed in the Laboratory Animal Facility of the Stanford University Medical Center. BALB/csuggested: RRID:IMSR_ORNL:BALB/cRl)Software and Algorithms Sentences Resources Cells were washed, fixed with 2% formaldehyde and acquired on a BD LSR II and analyzed using Cytobank V7.3.0. Cytobanksuggested: (Cytobank, RRID:SCR_014043)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: Please consider improving the rainbow (“jet”) colormap(s) used on page 3. At least one figure is not accessible to readers with colorblindness and/or is not true to the data, i.e. not perceptually uniform.
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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