The neutralization potency of anti-SARS-CoV-2 therapeutic human monoclonal antibodies is retained against novel viral variants

  1. Efi Makdasi
  2. Anat Zvi
  3. Ron Alcalay
  4. Tal Noy-Porat
  5. Eldar Peretz
  6. Adva Mechaly
  7. Yinon Levy
  8. Eyal Epstein
  9. Theodor Chitlaru
  10. Ariel Tennenhouse
  11. Moshe Aftalion
  12. David Gur
  13. Nir Paran
  14. Hadas Tamir
  15. Oren Zimhony
  16. Shay Weiss
  17. Michal Mandelboim
  18. Ella Mendelson
  19. Neta Zuckerman
  20. Ital Nemet
  21. Limor Kliker
  22. Shmuel Yitzhaki
  23. Shmuel C. Shapira
  24. Tomer Israely
  25. Sarel J. Fleishman
  26. Ohad Mazor
  27. Ronit Rosenfeld

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Abstract

A wide range of SARS-CoV-2 neutralizing monoclonal antibodies (mAbs) were reported to date, most of which target the spike glycoprotein and in particular its receptor binding domain (RBD) and N-terminal domain (NTD) of the S1 subunit. The therapeutic implementation of these antibodies has been recently challenged by emerging SARS-CoV-2 variants that harbor extensively mutated spike versions. Consequently, the re-assessment of mAbs, previously reported to neutralize the original early-version of the virus, is of high priority.

Four previously selected mAbs targeting non-overlapping epitopes, were evaluated for their binding potency to RBD versions harboring individual mutations at spike positions 417, 439, 453, 477, 484 and 501. Mutations at these positions represent the prevailing worldwide distributed modifications of the RBD, previously reported to mediate escape from antibody neutralization. Additionally, the in vitro neutralization potencies of the four RBD-specific mAbs, as well as two NTD-specific mAbs, were evaluated against two frequent SARS-CoV-2 variants of concern (VOCs): (i) the B.1.1.7 variant, emerged in the UK and (ii) the B.1.351 variant, emerged in South Africa. Variant B.1.351 was previously suggested to escape many therapeutic mAbs, including those authorized for clinical use. The possible impact of RBD mutations on recognition by mAbs is addressed by comparative structural modelling. Finally, we demonstrate the therapeutic potential of three selected mAbs by treatment of K18-hACE2 transgenic mice two days post infection with each of the virus strains.

Our results clearly indicate that despite the accumulation of spike mutations, some neutralizing mAbs preserve their potency against SARS-CoV-2. In particular, the highly potent MD65 and BL6 mAbs are shown to retain their ability to bind the prevalent novel viral mutations and to effectively protect against B.1.1.7 and B.1.351 variants of high clinical concern.

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  1. ScreenIT

    SciScore for 10.1101/2021.04.01.438035: (What is this?)

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

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Antibodies
    SentencesResources
    All antibodies (except LY-CoV555) were produced as full IgG1 antibodies as described (Barlev-Gross et al., 2021; Rosenfeld et al., 2021), expressed using ExpiCHO™ Expression system (Thermoscientific, USA) and purified on HiTrap Protein-A column (GE healthcare, UK).
    IgG1
    suggested: None
    For assessment of binding to S1 variants or mutated RBD, antibodies were captured on Protein-A or anti-Fab CH1 sensors (FAB2G) and incubated with recombinant S1 (WT, B.1.1.7 or B.1.351) or recombinant RBD (WT or mutated) for 180 sec and then transferred to buffer containing wells for additional 60 sec.
    anti-Fab
    suggested: None
    For epitope binning, MD65 antibody was biotinylated, immobilized on streptavidin sensor, incubated with a fixed concentration of WT rS1 (20 μg/ml) to reach saturation, washed and incubated with non-labeled LY-CoV555 for 180 sec.
    MD65
    suggested: None
    Experimental Models: Cell Lines
    SentencesResources
    Isolation and characterization of the MD29, MD65 and MD62 mAbs, targeting epitopes I-III on the RBD as previously reported.
    MD65
    suggested: RRID:CVCL_MD65)
    MD29 and MD65 were used as positive and negative controls, respectively.
    MD29
    suggested: RRID:CVCL_MD29)
    Titer of stock was determined by plaque assay using Vero E6 cells.
    Vero E6
    suggested: RRID:CVCL_XD71)
    Software and Algorithms
    SentencesResources
    The following His-tagged recombinant proteins were purchased from Sino Biologicals: B.1.1.7 rS1-SARS-CoV-2 spike S1 [Δ 69-70; Δ 144; N501Y; A570D; D614G; P681H], cat#40591-V08H12; B.1.351 rS1-SARS-CoV-2 spike S1 [K417N; E484K; N501Y; D614G], cat#40591-V08H10; spike RBD[N501Y] cat#40592-V08H82; spike RBD[S477N] cat#40592-V08H46; spike RBD[E484K] cat#40592-V08H84; spike RBD[N439K] cat#40592-V08H14; spike RBD[K417N] cat#40592-V08H59; spike RBD[Y453F] cat#40592-V08H80.
    Sino Biologicals
    suggested: None
    Area under curve (AUC) was calculated for each binding curve, using GraphPad Prism 5, and percent binding was calculated compared to the WT protein, representing 100% binding.
    GraphPad Prism
    suggested: (GraphPad Prism, RRID:SCR_002798)
    The identity of the B.1.1.7 strain was confirmed using NGS.
    NGS
    suggested: (PM4NGS, RRID:SCR_019164)

    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.

    About SciScore

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  2. Version published to 10.1101/2021.04.01.438035 on bioRxiv