tetano
Editor, Senior Moderator
Adv Biol (Weinh)
. 2021 Oct 27;e2101113.
doi: 10.1002/adbi.202101113. Online ahead of print.
NMR-Based Analysis of Nanobodies to SARS-CoV-2 Nsp9 Reveals a Possible Antiviral Strategy Against COVID-19
Gennaro Esposito[SUP] 1 2 [/SUP], Yamanappa Hunashal[SUP] 1 [/SUP], Mathias Percipalle[SUP] 1 3 [/SUP], Tomas Venit[SUP] 4 [/SUP], Mame Massar Dieng[SUP] 4 [/SUP], Federico Fogolari[SUP] 2 5 [/SUP], Gholamreza Hassanzadeh[SUP] 6 [/SUP], Fabio Piano[SUP] 4 7 [/SUP], Kristin C Gunsalus[SUP] 8 9 [/SUP], Youssef Idaghdour[SUP] 4 7 [/SUP], Piergiorgio Percipalle[SUP] 4 10 [/SUP]
Affiliations
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.
. 2021 Oct 27;e2101113.
doi: 10.1002/adbi.202101113. Online ahead of print.
NMR-Based Analysis of Nanobodies to SARS-CoV-2 Nsp9 Reveals a Possible Antiviral Strategy Against COVID-19
Gennaro Esposito[SUP] 1 2 [/SUP], Yamanappa Hunashal[SUP] 1 [/SUP], Mathias Percipalle[SUP] 1 3 [/SUP], Tomas Venit[SUP] 4 [/SUP], Mame Massar Dieng[SUP] 4 [/SUP], Federico Fogolari[SUP] 2 5 [/SUP], Gholamreza Hassanzadeh[SUP] 6 [/SUP], Fabio Piano[SUP] 4 7 [/SUP], Kristin C Gunsalus[SUP] 8 9 [/SUP], Youssef Idaghdour[SUP] 4 7 [/SUP], Piergiorgio Percipalle[SUP] 4 10 [/SUP]
Affiliations
- PMID: 34705339
- DOI: 10.1002/adbi.202101113
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.