Broadly neutralizing antibodies against Omicron variants of SARS-CoV-2 derived from mRNA-lipid nanoparticle-immunized mice
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Abstract
The COVID-19 pandemic continues to threaten human health worldwide, as new variants of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have emerged. Currently, the predominant circulating strains around the world are Omicron variants, which can evade many therapeutic antibodies. Thus, the development of new broadly neutralizing antibodies remains an urgent need. In this work, we address this need by using the mRNA-lipid nanoparticle immunization method to generate a set of Omicron-targeting monoclonal antibodies. Five of our novel K-RBD-mAbs show strong binding and neutralizing activities toward all SARS-CoV-2 variants of concern (Alpha, Beta, Gamma, Delta and Omicron). Notably, the epitopes of these five K-RBD-mAbs are overlapping and localized around K417 and F486 of the spike protein receptor binding domain (RBD). Chimeric derivatives of the five antibodies (K-RBD-chAbs) neutralize Omicron sublineages BA.1 and BA.2 with low IC 50 values that range from 5.7 to 12.9 ng/mL. Additionally, we performed antibody humanization on a broadly neutralizing chimeric antibody to create K-RBD-hAb-62, which still retains excellent neutralizing activity against Omicron. Our results collectively suggest that these five therapeutic antibodies may effectively combat current and emerging SARS-CoV-2 variants, including Omicron BA.1 and BA.2. Therefore, the antibodies can potentially be used as universal neutralizing antibodies against SARS-CoV-2.
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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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