An ultrapotent neutralizing bispecific antibody with broad spectrum against SARS-CoV-2 variants

  1. Hui Zhang
  2. Haohui Huang
  3. Rong Li
  4. Lu Zhang
  5. Zhiwei Wang
  6. Jiaping Li
  7. Junyou Chen
  8. Huafei Su
  9. Dandan Zheng
  10. Ziqi Su
  11. Li Wang
  12. Chunping Deng
  13. shujun Pei
  14. Shenghua Zhu
  15. Chan Li
  16. Yaochang Yuan
  17. Haitao Yue
  18. Yanqun Wang
  19. Xiaobo Li
  20. Cuihua Liu
  21. Jinchen Yu
  22. Hui Zhang
  23. Shengfeng Li
  24. Xianming Huang

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Abstract

In spite of the successful development of effective countermeasures against Covid-19, variants have and will continue to emerge that could compromise the efficacy of currently approved neutralizing antibodies and vaccines. Consequently, novel and more efficacious agents are urgently needed. We have developed a bispecific antibody, 2022, consisting of two antibodies, 2F8 and VHH18. 2F8 was isolated from our proprietary fully synthetic human IDEAL (Intelligently Designed and Engineered Antibody Library)-VH/VL library and VHH18 is a single domain antibody isolated from IDEAL-nanobody library. 2022 was constructed by attaching VHH18 to the C-terminal of Fc of 2F8. 2022 binds two non-overlapping epitopes simultaneously on the RBD of the SARS-CoV-2 spike protein and blocks the binding of RBD to human angiotensin-converting enzyme 2 (ACE2). 2022 potently neutralizes SARS-CoV-2 and all of the variants tested in both pseudovirus and live virus assays, including variants carrying mutations known to resist neutralizing antibodies approved under EUA and that reduce the protection efficiency of current effective vaccines. The half-maximum inhibitory concentration (IC50) of 2022 is 270 pM, 30 pM, 20 pM, and 1 pM, for wild-type, alpha, beta, and delta pseudovirus, respectively. In the live virus assay, 2022 has an IC50 of 26.4 pM, 13.3 pM, and 88.6 pM, for wild-type, beta, and delta live virus, respectively. In a mouse model of SARS-CoV-2, 2022 showed strong prophylactic and therapeutic effects. A single administration of 2022 intranasal (i.n.) or intraperitoneal (i.p.) 24 hours before virus challenge completely protected all mice from bodyweight loss, as compared with up to 20% loss of bodyweight in placebo treated mice. In addition, the lung viral titers were undetectable (FRNT assay) in all mice treated with 2022 either prophylactically or therapeutically, as compared with around 1×10 5 pfu/g lung tissue in placebo treated mice. In summary, bispecific antibody 2022 showed potent binding and neutralizing activity across a variety of SARS-CoV-2 variants and could be an attractive weapon to combat the ongoing waves of the COVID-19 pandemic propagated mainly by variants, especially, the much more contagious delta variant.

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  1. ScreenIT

    SciScore for 10.1101/2021.08.10.455627: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    EthicsIACUC: All protocols were approved by the Institutional Animal Care and Use Committees of Guangzhou Customs District Technology Center and Guangzhou Medical University.
    Sex as a biological variablenot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    Bound antibodies were detected with Peroxidase conjugated goat anti-human kappa light chains antibody (A7164-1ML, Sigma).
    anti-human kappa light chains
    suggested: None
    Affinity measurement of antibodies by Surface plasmon resonance (SPR): SPR experiments were all conducted with a Biacore T200 system (GE Healthcare); All assays were performed with a Sensor Chip Protein A (GE healthcare),with a HBS EP+ running buffer (0.1M HEPES, 1.5M NaCl, 0.03M EDTA supplemented with 0.005% vol/vol surfactant P20 at 25°C.) To determine the affinities of nanobody VHH18, human IgG1 antibody 2F8, and 2022 to SARS-CoV-2 RBD-His tag, spike trimer-His tag and other S1-His tag variants, antibodies were immobilized onto the sample flow paths of the sensor Protein A chip.
    human IgG1
    suggested: None
    The sensor was saturated with the first antibody, either 2F8 Fab or VHH18-His, subsequently, the above bioprobes were flown over with the different second antibody, either VHH-his or 2F8 Fab, respectively.
    VHH18-His
    suggested: None
    Cells were tested with a rabbit anti-SARS-CoV-2 nucleocapsid protein polyclonal antibody (Cat. No.: 40143-T62, SinoBiological, Inc.), and an HRP-labelled goat anti-rabbit as secondary antibody (111-035-003, Jackson ImmunoResearch).
    anti-SARS-CoV-2 nucleocapsid protein
    suggested: (Proteintech Cat# 67666-1-Ig, RRID:AB_2882862)
    anti-rabbit
    suggested: (Jackson ImmunoResearch Labs Cat# 111-035-003, RRID:AB_2313567)
    Experimental Models: Cell Lines
    SentencesResources
    Cell lines: HEK293 (ACS-4500TM, ATCC) and African Green monkey kidney-derived VeroE6 cells (CRL-1587, ATCC) were cultured and passaged in DMEM with 10% FBS.
    ACS-4500TM
    suggested: None
    VeroE6
    suggested: JCRB Cat# JCRB1819, RRID:CVCL_YQ49)
    Transiently transfected to HEK293 cells to obtain recombinant RBD alanine mutants and purified by protein A columns.
    HEK293
    suggested: CLS Cat# 300192/p777_HEK293, RRID:CVCL_0045)
    Experimental Models: Organisms/Strains
    SentencesResources
    HEK293-hACE2 was constructed and sorted by Bio-Thera Solutions, Ltd. Recombinant Proteins: Biotinylated 2019-nCoV S protein RBD, His,Avitag™ (SPD-C82E9,Acrobiosystems); SARS-Vov-2 S protein RBD, His Tag (SPD-S52H6,
    S protein RBD
    suggested: None
    Balb/c mice were mildly anesthetized with isoflurane and i.n. transduced with 2.5×109 PFU of Ad5-ACE2 in 75ul DMEM.
    Balb/c
    suggested: None
    Software and Algorithms
    SentencesResources
    Values were determined using four-parameter logistic regression (GraphPad Prism).
    GraphPad
    suggested: (GraphPad Prism, RRID:SCR_002798)

    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.
    • No funding statement was detected.
    • 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.

  2. Version published to 10.1101/2021.08.10.455627v1 on bioRxiv