ORIGINAL RESEARCH article
Front. Immunol.
Sec. Viral Immunology
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1585793
This article is part of the Research TopicAntiviral Innate Immune Mechanisms in Animal HostsView all 4 articles
Development of a potent neutralizing nanobody against canine distemper virus hemagglutinin protein
Provisionally accepted
- China Agricultural University, Beijing, China
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Canine distemper virus (CDV) is the etiological agent of canine distemper. The virus can infect canids irrespective of age, sex, or breed, leading to a highly contagious and lethal disease that seriously threatens the health of canids, fur animals, and wildlife. Although vaccination can currently prevent CDV infection, developing effective emergency treatment drugs remains crucial. Nanobodies derived from camelid or shark heavy chain-only antibodies can effectively inhibit viral infections, suggesting their potential as therapeutic agents for treating CDV infection. In this study, we utilized a phage display nanobody library constructed from immunized alpacas and isolated a nanobody (Nb-6C6) that specifically binds to the CDV hemagglutinin (H) protein. Nb-6C6 was successfully expressed in mammalian cells and exhibited high binding affinity to CDV H (EC50 = 0.174 µg/mL).Neutralization assays further revealed that Nb-6C6 could effectively neutralize CDV (IC50 = 1.773 µg/mL). Fusion of Nb-6C6 with canine IgG Fc resulted in homodimers, significantly increasing its neutralizing activity by up to 4.6-fold. AlphaFold3 analysis indicated that the neutralizing capacity of Nb-6C6 against CDV is attributed to an interaction between residue D106 in the CDR3 region and the conserved residue R408 of the H protein. These findings suggest that the nanobody Nb-6C6 and its bivalent form exhibit high-affinity binding and potent neutralizing activity against CDV, highlighting their potential as promising therapeutic candidates for the treatment of CDV infection.
Keywords: Canine distemper virus, Hemagglutinin protein, Nanobody, phage display, neutralizing antibody
Received: 01 Mar 2025; Accepted: 15 Apr 2025.
Copyright: © 2025 Xiao, Wu, Su and Wu. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Jingliang Su, China Agricultural University, Beijing, China
Qingmin Wu, China Agricultural University, Beijing, China
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