Broadly neutralizing antibodies against Omicron variants of SARS-CoV-2 derived from mRNA-lipid nanoparticle-immunized mice

  1. Ruei-Min Lu
  2. Kang-Hao Liang
  3. Hsiao-Ling Chiang
  4. Fu-Fei Hsu
  5. Hsiu-Ting Lin
  6. Wan-Yu Chen
  7. Feng-Yi Ke
  8. Monika Kumari
  9. Yu-Chi Chou
  10. Mi-Hua Tao
  11. Yi-Ling Lin
  12. Han-Chung Wu

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Abstract

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  1. Version published to 10.1016/j.heliyon.2023.e15587
  4. ScreenIT

    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

    EthicsIACUC: All animal experiments were approved by the Academia Sinica Institutional Animal Care and Use Committee (IACUC protocol No.
    Sex as a biological variableFour- 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.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Anti-RBD and control antibodies were added to the plates and incubated for 1 h at room temperature.
    Anti-RBD
    suggested: None
    Then, the cells were washed and horseradish peroxidase-conjugated anti-human antibody (1:2000) was added for 1 h at room temperature.
    anti-human antibody
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    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-1
    suggested: RRID:CVCL_IV58)
    The pseudovirus neutralization assays were performed using HEK293T cells that expressed human ACE2 (HEK293T/hACE2)
    HEK293T
    suggested: None
    The mixtures were then added to pre-seeded HEK293T/hACE2 cells for 24 h at 37°C.
    HEK293T/hACE2
    suggested: RRID:CVCL_A7UK)
    Experimental Models: Organisms/Strains
    SentencesResources
    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/c
    suggested: RRID:IMSR_ORNL:BALB/cRl)
    Recombinant DNA
    SentencesResources
    The PCR products were cloned using the pGEM-T Easy Vector System (Promega) and analyzed by DNA sequencing.
    pGEM-T Easy
    suggested: RRID:Addgene_86229)
    The resulting VH was cloned into a modified pcDNA5-FRT-Gamma1 expression vector with human IgG1 constant region.
    pcDNA5-FRT-Gamma1
    suggested: None
    Software and Algorithms
    SentencesResources
    Recombinant protein-based ELISA: Recombinant RBD and spike-His tag proteins for different SARS-CoV-2 variants were purchased from ACROBiosystems.
    ACROBiosystems
    suggested: (ACRObiosystems, RRID:SCR_012550)
    The half maximal inhibitory concentration (IC50) was calculated by nonlinear regression using Prism software version 8.1.0 (
    Prism
    suggested: (PRISM, RRID:SCR_005375)
    GraphPad Software Inc.)
    GraphPad
    suggested: (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/).
    IgBLAST
    suggested: (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.
    PyMOL
    suggested: (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.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.

  5. Version published to 10.1101/2022.04.19.488843v1 on bioRxiv