NMR-Based Analysis of Nanobodies to SARS-CoV-2 Nsp9 Reveals a Possible Antiviral Strategy Against COVID-19

Gennaro Esposito, Yamanappa Hunashal, Mathias Percipalle, Tomas Venit, Mame Massar Dieng, Federico Fogolari, Gholamreza Hassanzadeh, Fabio Piano, Kristin C. Gunsalus, Youssef Idaghdour, Piergiorgio Percipalle

Research output: Contribution to journalArticlepeer-review


Following the entry into the host cell, SARS-CoV-2 replication is mediated by the replication transcription complex (RTC) assembled through a number of nonstructural proteins (Nsps). A monomeric form of Nsp9 is particularly important for RTC assembly and function. In the present study, 136 unique nanobodies targeting Nsp9 are generated. Several nanobodies belonging to different B-cell lineages are expressed, purified, and characterized. Results from immunoassays applied to purified Nsp9 and neat saliva from coronavirus disease (COVID-19) patients show that these nanobodies effectively and specifically recognize both recombinant and endogenous Nsp9. Nuclear magnetic resonance analyses supported by molecular dynamics reveal a composite Nsp9 oligomerization pattern and demonstrate that both nanobodies stabilize the tetrameric form of wild-type Nsp9 also identifying the epitopes on the tetrameric assembly. These results can have important implications in the potential use of these nanobodies to combat viral replication.

Original languageEnglish (US)
Article number2101113
JournalAdvanced Biology
Issue number12
StatePublished - Dec 2021


  • COVID-19
  • NMR spectroscopy
  • Nsp9
  • SARS-CoV-2
  • nanobodies

ASJC Scopus subject areas

  • General Biochemistry, Genetics and Molecular Biology
  • Biomedical Engineering
  • Biomaterials
  • General Medicine


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