Supplementary Materials

Supplementary Material for:

A multimechanistic antibody targeting the receptor binding site potently cross-protects against influenza B viruses

Chenguang Shen, Junyu Chen, Rui Li, Mengya Zhang, Guosong Wang, Svetlana Stegalkina, Limin Zhang, Jing Chen, Jianli Cao, Xingjian Bi, Stephen F. Anderson, Timothy Alefantis, Minwei Zhang, Xiaoyang Cai, Kunyu Yang, Qingbing Zheng, Mujing Fang, Hai Yu, Wenxin Luo, Zizheng Zheng, Quan Yuan, Jun Zhang, James Wai-Kuo Shih, Harry Kleanthous, Honglin Chen, Yixin Chen,*
Ningshao Xia*

*Corresponding author. Email: yxchen2008{at}xmu.edu.cn (Y.C.); nsxia{at}xmu.edu.cn (N.X.)

Published 18 October 2017, Sci. Transl. Med. 9,eaam5752 (2017)
DOI: 10.1126/scitranslmed.aam5752

This PDF file includes:

  • Materials and Methods
  • Fig. S1. A schematic diagram showing the 12G6 generation and selection process.
  • Fig. S2. In vitro binding and neutralization activities of mouse antibody 12G6.
  • Fig. S3. Nucleotide and amino acid sequences of the VH and VL chain regions of C12G6.
  • Fig. S4. Binding curves for reported Kd values for binding of C12G6 to influenza B HAs.
  • Fig. S5. Immunofluorescence assay activity of C12G6 against both lineages of influenza B viruses.
  • Fig. S6. Binding of C12G6 to MDCK cells infected with influenza B viruses, detected by flow cytometry.
  • Fig. S7. C12G6 binds the B/Florida/4/2006 HA subunit in Western blotting.
  • Fig. S8. Expression, purification, and characterization of influenza B HA bnAbs.
  • Fig. S9. In vitro neutralization activities (IC50 values) of C12G6 and other reported influenza B HA bnAbs.
  • Fig. S10. Body weight change curves of mice treated with C12G6 before or after challenge with influenza B viruses.
  • Fig. S11. Immunofluorescence assay activity of anti–influenza B nucleoprotein mAb 4D5 against both lineages of influenza B viruses.
  • Fig. S12. Histological analysis of lungs from mice prophylactically treated with C12G6 before infection with influenza B viruses.
  • Fig. S13. Histological analysis of lungs from mice therapeutically treated with C12G6 after infection with influenza B viruses.
  • Fig. S14. Comparison of therapeutic effects of C12G6 and oseltamivir against influenza B infection in mice.
  • Fig. S15. Body weight change curves of ferrets treated with C12G6 before or after challenge with influenza B viruses.
  • Fig. S16. Clinical sign of ferrets after prophylactic and therapeutic treatment with C12G6 against influenza B infection.
  • Fig. S17. In vivo fitness of mutant B/Hong Kong/537/2009–like viruses.
  • Fig. S18. Reactivity of C12G6 with WT or mutant B/Hong Kong/537/2009–like viruses in ELISA.
  • Fig. S19. C12G6 does not block HA0 activation.
  • Fig. S20. C12G6 inhibits syncytia formation.
  • Fig. S21. Red blood cell fusion assay.
  • Fig. S22. Comparison of the in vitro binding and neutralizing properties of C12G6 and C12G6-LALA.
  • Fig. S23. Serum antibody concentrations after C12G6 or C12G6-LALA treatment.
  • Fig. S24. ADCC and CDC activities of C12G6 against the B/Lee/1940 virus strain.
  • Fig. S25. Determination of the role of antibody effector functions in the protective efficacy of C12G6.
  • Fig. S26. ADCP activity of 12G6 against influenza B viruses.
  • Fig. S27. A schematic diagram showing the inhibition mechanisms of C12G6.
  • Table S1. ELISA EC50 and SD values of mouse antibodies 12G6 and 5G6, related to fig. S2.
  • Table S2. The full designations and abbreviations of the influenza virus strains used in this study.
  • Table S3. ELISA EC50 and SD values of C12G6 and C5G6 antibodies, related to Fig. 1B.
  • Table S4. Kd for binding of C12G6 to HAs of influenza B strains.
  • Table S5. ELISA EC50 and SD values of the indicated antibodies, related to fig. S8B.
  • Table S6. Neutralization IC50 and SD values of C12G6 and CR8033-like antibodies, related to Fig. 1D.
  • Table S7. Neutralization IC50 and SD values of CR8071-like and CR9114-like antibodies, related to Fig. 1D.
  • Table S8. Neutralization IC50 and SD values of 5A7-like and C5G6 antibodies, related to Fig. 1D.
  • Table S9. Characterization of interaction surface area.
  • Table S10. ELISA EC50 and SD values of C12G6 and C12G6-LALA antibodies, related to fig. S22.
  • Table S11. C12G6 reactivity pattern.
  • Table S12. HA sequences of influenza B viruses used in this study.
  • References (27–33)

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Other Supplementary Material for this manuscript includes the following:

  • Table S13 (Microsoft Excel format). Primary data.