Gu M, Jiao J, Liu S, Zhao W, Ge Z, Cai K, Xu L, He. Monoclonal antibody targeting a novel linear epitope on nucleoprotein confers pan-reactivity to influenza A virus. Appl Microbiol Biotechnol. 2023 Feb 23:1–14.
Nucleoprotein (NP) functions crucially in the replicative cycle of influenza A virus (IAV) via forming the ribonucleoprotein complex together with PB2, PB1, and PA proteins. As its high conservation, NP ranks one of the hot targets for design of universal diagnostic reagents and antiviral drugs for IAV. Here, we report an anti-NP murine monoclonal antibody (mAb) 5F10 prepared from traditional lymphocyte hybridoma technique with the immunogen of a clade 2.3.4.4 H5N1 subtype avian influenza virus. The specificity of mAb 5F10 to NP protein was confirmed by immunofluorescence assay and western blotting, and the mAb 5F10 could be used in immunoprecipitation and immunohistochemistry assays. Importantly, mAb 5F10 possessed broad-spectrum reactivity against H1~H11 subtypes of avian influenza viruses, including various HA clades of H5Nx subtype. In addition, mAb 5F10 also showed good affinity with H1N1 and H3N2 subtype influenza viruses of swine and human origin. Furthermore, the recognized antigenic epitope of mAb 5F10 was identified to consist of the conserved amino acid motif 81EHPSA85 in the second flexible loop region of NP protein through screening the phage display peptide library. Collectively, the mAb 5F10 which recognizes the novel universal NP linear B-cell epitope of IAV with diverse origins and subtypes will be a powerful tool for NP protein-based structural, functional, and mechanistic studies, as well as the development of detection methods and universal vaccines for IAV. KEY POINTS: ? A broad-spectrum mAb against various subtypes and sources of IAV was developed ? The mAb possessed good reactivity in IFA, western blot, IP, and IHC assays ? The mAb targeted a novel conserved linear B-cell epitope involving 81EHPSA85 on NP protein.
See Also:
Latest articles in those days:
- An aggregated dataset of serial morbidity and titer measurements from influenza A virus-infected ferrets 22 hours ago
- Structures of H5N1 influenza polymerase with ANP32B reveal mechanisms of genome replication and host adaptation 4 days ago
- Risk assessment of a highly pathogenic H5N1 influenza virus from mink 4 days ago
- Detection of clade 2.3.4.4b highly pathogenic H5N1 influenza virus in New York City 4 days ago
- Sequence-based epitope mapping of high pathogenicity avian influenza H5 clade 2.3.4.4b in Latin America 4 days ago
[Go Top] [Close Window]