@article{0e5ff92168db4d9d8c46c8975599e902,
title = "NMR-Based Analysis of Nanobodies to SARS-CoV-2 Nsp9 Reveals a Possible Antiviral Strategy Against COVID-19",
abstract = "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.",
keywords = "COVID-19, NMR spectroscopy, Nsp9, SARS-CoV-2, nanobodies",
author = "Gennaro Esposito and Yamanappa Hunashal and Mathias Percipalle and Tomas Venit and Dieng, {Mame Massar} and Federico Fogolari and Gholamreza Hassanzadeh and Fabio Piano and Gunsalus, {Kristin C.} and Youssef Idaghdour and Piergiorgio Percipalle",
note = "Funding Information: The authors thank Xin Xie, NYUAD Center for Genomics and Systems Biology, and NYUAD CTP for the use of the instrumentation. The authors also thank the COVID-19 consortium and in particular Prof. A. Pastore for sharing the wild-type Nsp9 expression protocol. This work was supported by NYUAD research funds to GE and PP, a COVID-19 Facilitator Grant by New York University Abu Dhabi and a donation from the Cipriani family to PP. Immunization and handling of the llama were performed according to directive 2010/63/EU of the European parliament for the protection of animals used for scientific purposes and approved by the Ethical Committee for Animal Experiments of the Vrije Universiteit Brussel (permit No. 13-601-2), Brussels, Belgium. Saliva samples from COVID-19 patients were collected under NYUAD IRB-approved protocol HRPP-2020-48 (PI Idaghdour). Saliva collections were performed under informed consent of all participating subjects. Funding Information: The authors thank Xin Xie, NYUAD Center for Genomics and Systems Biology, and NYUAD CTP for the use of the instrumentation. The authors also thank the COVID‐19 consortium and in particular Prof. A. Pastore for sharing the wild‐type Nsp9 expression protocol. This work was supported by NYUAD research funds to GE and PP, a COVID‐19 Facilitator Grant by New York University Abu Dhabi and a donation from the Cipriani family to PP. Immunization and handling of the llama were performed according to directive 2010/63/EU of the European parliament for the protection of animals used for scientific purposes and approved by the Ethical Committee for Animal Experiments of the Vrije Universiteit Brussel (permit No. 13‐601‐2), Brussels, Belgium. Saliva samples from COVID‐19 patients were collected under NYUAD IRB‐approved protocol HRPP‐2020‐48 (PI Idaghdour). Saliva collections were performed under informed consent of all participating subjects. Publisher Copyright: {\textcopyright} The Authors. Advanced Biology published by Wiley-VCH GmbH",
year = "2021",
month = dec,
doi = "10.1002/adbi.202101113",
language = "English (US)",
volume = "5",
journal = "Advanced Biology",
issn = "2701-0198",
publisher = "Wiley-VCH Verlag",
number = "12",
}