1College
of Zoology, Gansu Agricultural University, Lanzhou, People's
Republic of China.2China Agricultural Vet. Bio.
Science and Technology Co. LTD, Lanzhou, People's Republic of China
*Corresponding author:
zengqy@gsau.edu.cn
Abstract
Previously, nanobodies (Nbs) targeting ORF2 protein of novel goose astrovirus (nGAstV)
were identified and tested for their specificity and binding affinity to nGAstV.
However, their ability to neutralize and prevent the viral infection in goslings
remained unexplored, especially those fused with Fc domain of chicken IgG (cFc).
In the present study, using nGAstV ORF2 as target antigen, Nb-positive clones
from already developed Nb phage library reacting with nGAstV were obtained
through three rounds of panning and enrichment. Four Nbs based on sequence
variation and solubility were selected, fused with cFc, and genetically
engineered to produce chimeric nanobodies. The specific reactivity of these
Nbs-Fc with nGAstV was assessed by IFA and WB analysis, the affinity and IC50
of Nbs-Fc were determined by bio-layer Interferometry tests and iELISA,
respectively. And the neutralizing activity of one nanobody with good biological
activity (Nb9-2-cFc) was assessed by cell infection and in vivo antiviral
assays in goslings. All the four Nbs-Fc were successfully expressed and
confirmed through SDS-PAGE. These Nbs exhibited strong binding affinity to
nGAstV, as demonstrated by IFA and WB analysis. Among these, Nb9-2-cFc showed
the highest affinity (Kd=59.6 pM) and neutralization activity
(IC50=15.75ng/mL) against nGAstV. Further, In vitro assays
revealed that this Nb could reduce virus proliferation by blocking the binding
of ORF2 to its receptor. In vivo evaluation of prophylactic activity in
goslings revealed that Nb9-2-cFc provided significant protection against nGAstV
infection by reducing viral load and histopathological damage in target organs.
These findings highlight Nb9-2-cFc as a promising candidate for emergency
prevention and treatment against nGAstV infection in goslings.
To Cite This Article:
Ge Y, Yang J, Liu X, An F, Cui Z and Zeng Q,
2025.
Single-domain antibody fused with chicken IgG Fc-fragment protects goslings
against novel goose astrovirus (nGAstV).
Pak Vet J. http://dx.doi.org/10.29261/pakvetj/2025.120