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Discovery of nanobodies against SARS-CoV-2 and an uncommon neutralizing mechanism
Dianfan Li; Tingting Li; Bingjie Zhou; Zhipu Luo; Yanling Lai; Suqiong Huang; Yuanze Zhou; Anupriya Gautam; Salome Bourgeau; Shurui Wang; Juan Bao; Jingquan Tan; Dimitri Lavillette.
Afiliación
  • Dianfan Li; Chinese Academy of Sciences
  • Tingting Li; CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (CAS)
  • Bingjie Zhou; CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai CAS
  • Zhipu Luo; Soochow University
  • Yanling Lai; CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (CAS)
  • Suqiong Huang; CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai CAS
  • Yuanze Zhou; Nanjing Crycision Biotech Co., Ltd.
  • Anupriya Gautam; CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai CAS
  • Salome Bourgeau; CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai CAS
  • Shurui Wang; Nanjing Crycision Biotech Co., Ltd
  • Juan Bao; State Key Laboratory of Molecular Biology, CAS Center for Excellence in Molecular Cell Science, Shanghai Institute of Biochemistry and Cell Biology
  • Jingquan Tan; Nanjing Crycision Biotech Co., Ltd
  • Dimitri Lavillette; Institut Pasteur Shanghai Chine Academy of Sciences
Preprint en En | PREPRINT-BIORXIV | ID: ppbiorxiv-453054
ABSTRACT
SARS-CoV-2 and its variants continue to threaten public health. The virus recognizes the host cell by attaching its Spike receptor-binding domain (RBD) to the host receptor ACE2. Therefore, RBD is a primary target for neutralizing antibodies and vaccines. Here we report the isolation, and biological and structural characterization of two single-chain antibodies (nanobodies, DL4 and DL28) from RBD-immunized alpaca. Both nanobodies bind Spike with affinities that exceeded the detection limit (picomolar) of the biolayer interferometry assay and neutralize the original SARS-CoV- 2 strain with IC50 of 0.086 g mL-1 (DL4) and 0.385 g mL-1 (DL28). DL4 and a more potent, rationally designed mutant, neutralizes the Alpha variant as potently as the original strain but only displays marginal activity against the Beta variant. By contrast, the neutralizing activity of DL28, when in the Fc-fused divalent form, was less affected by the mutations in the Beta variant (IC50 of 0.414 g mL-1 for Alpha, 1.060 g mL-1 for Beta). Crystal structure studies reveal that DL4 blocks ACE2-binding by direct competition, while DL28 neutralizes SARS-CoV-2 by an uncommon mechanism through which DL28 distorts the receptor-binding motif in RBD and hence prevents ACE2-binding. Our work provides two neutralizing nanobodies for potential therapeutic development and reveals an uncommon mechanism to design and screen novel neutralizing antibodies against SARS-CoV-2.
Licencia
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Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-BIORXIV Idioma: En Año: 2021 Tipo del documento: Preprint
Texto completo: 1 Colección: 09-preprints Base de datos: PREPRINT-BIORXIV Idioma: En Año: 2021 Tipo del documento: Preprint