Broadly neutralizing antibodies against Omicron variants of SARS-CoV-2 derived from mRNA-lipid nanoparticle-immunized mice
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Juan Pablo Jaworski
Review of "Broadly Neutralizing antibodies against Omicron variants of SARS-CoV-2 derived from mRNA-lipid nanoparticle-immunized mice"
Reviewer: Juan Pablo Jaworski (IVIT - CONICET) | 📗📗📗📗◻️
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Juan Pablo Jaworski
Review 1: "Broadly Neutralizing antibodies against Omicron variants of SARS-CoV-2 derived from mRNA-lipid nanoparticle-immunized mice"
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SciScore for 10.1101/2022.04.19.488843: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: All animal experiments were approved by the Academia Sinica Institutional Animal Care and Use Committee (IACUC protocol No. Sex as a biological variable Four- to six-week-old female BALB/c mice were immunized with 5 μg of the Kappa spike and RBD mRNA-LNP by intramuscular (I.M.) injection. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Anti-RBD and control antibodies were added to the plates and incubated for 1 h at room temperature. Anti-RBDsuggested: NoneThen, the cells were washed and horseradish peroxidase-conjugated anti-human antibody (1:2000) was added for 1 h at room … SciScore for 10.1101/2022.04.19.488843: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Ethics IACUC: All animal experiments were approved by the Academia Sinica Institutional Animal Care and Use Committee (IACUC protocol No. Sex as a biological variable Four- to six-week-old female BALB/c mice were immunized with 5 μg of the Kappa spike and RBD mRNA-LNP by intramuscular (I.M.) injection. Randomization not detected. Blinding not detected. Power Analysis not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Anti-RBD and control antibodies were added to the plates and incubated for 1 h at room temperature. Anti-RBDsuggested: NoneThen, the cells were washed and horseradish peroxidase-conjugated anti-human antibody (1:2000) was added for 1 h at room temperature. anti-human antibodysuggested: NoneExperimental Models: Cell Lines Sentences Resources After four inoculations with the same concentration of mRNA-LNP, the splenocytes from immunized mice were harvested and fused with mouse myeloma NS-1 cells. NS-1suggested: RRID:CVCL_IV58)The pseudovirus neutralization assays were performed using HEK293T cells that expressed human ACE2 (HEK293T/hACE2) HEK293Tsuggested: NoneThe mixtures were then added to pre-seeded HEK293T/hACE2 cells for 24 h at 37°C. HEK293T/hACE2suggested: RRID:CVCL_A7UK)Experimental Models: Organisms/Strains Sentences Resources Four- to six-week-old female BALB/c mice were immunized with 5 μg of the Kappa spike and RBD mRNA-LNP by intramuscular (I.M.) injection. BALB/csuggested: RRID:IMSR_ORNL:BALB/cRl)Recombinant DNA Sentences Resources The PCR products were cloned using the pGEM-T Easy Vector System (Promega) and analyzed by DNA sequencing. pGEM-T Easysuggested: RRID:Addgene_86229)The resulting VH was cloned into a modified pcDNA5-FRT-Gamma1 expression vector with human IgG1 constant region. pcDNA5-FRT-Gamma1suggested: NoneSoftware and Algorithms Sentences Resources Recombinant protein-based ELISA: Recombinant RBD and spike-His tag proteins for different SARS-CoV-2 variants were purchased from ACROBiosystems. ACROBiosystemssuggested: (ACRObiosystems, RRID:SCR_012550)The half maximal inhibitory concentration (IC50) was calculated by nonlinear regression using Prism software version 8.1.0 ( Prismsuggested: (PRISM, RRID:SCR_005375)GraphPad Software Inc.) GraphPadsuggested: (GraphPad Prism, RRID:SCR_002798)From the sequences, the framework regions (FRs) and complementarity determining regions (CDRs) were defined by searching with the NCBI IgBLAST program (https://www.ncbi.nlm.nih.gov/igblast/). IgBLASTsuggested: (IgBLAST, RRID:SCR_002873)After analyzing the structure with PyMOL software, we identified the key amino acid residues at which mutations may impact the original conformation of the CDRs. PyMOLsuggested: (PyMOL, RRID:SCR_000305)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.
Results from scite Reference Check: We found no unreliable references.
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