United Kingdom (UK)
Recombinant monoclonal antibody to Influenza A nucleoprotein. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from phage display aNP-VHH1.
UniProt Accession Number of Target Protein: P03466
Alternative Name(s) of Target: IAV NP; Influenza A virus nucleoprotein; Influenza A NP; Protein N; αNP-VHH1
Immunogen: The original nanobody was generated by subjecting alpacas to five rounds of immunization with alcohol-fixed influenza virus PR8. The generated phage library underwent three separate methods of panning to isolate influenza virus-specific VHHs.
Specificity: This antibody is specific for an epitope on the influenza virus A NP. The epitope is thought to overlap with a protein region responsible for several host protein interactions, including importin-a and Mx proteins. Immunoprecipitation confirmed no reactivity towards the M1, PB1, PB2, or PA proteins of Influenza A.
Application Notes: The original version of this antibody (alpaca VHH) was confirmed to target NP antigens in an immunoprecipitation assay. The specificity of this antibody's binding has been validated using both a His tag and mCherry attachment. Furthermore, X-ray crystallography revealed its binding with IVA NP, where it binds to the NP body domain. Mini-genome assays have shown that this antibody not only interferes with Influenza A replication but also prevents the nuclear import of RNPs. Additionally, the antibody has been utilized for the immunoprecipitation of IVA NP and in Western blotting, further confirming its inhibitory effect on the nuclear localization of its target ribonuclear protein, disrupting the virus's function (Ashour et al., 2015; PMID: 25540369). Flow cytometry, in conjunction with negative-stain EM, demonstrated the interference of influenza A vRNP nuclear import by the VHH version of this antibody. This interference occurs through the masking of the nuclear localization sequence of NPs, proposing a model for the inhibition of vRNP nuclear import (Hanke et al., 2016; PMID: 27965447).
Antibody first published in:
Ashour et al. Intracellular expression of camelid single-domain antibodies specific for influenza virus nucleoprotein uncovers distinct features of its nuclear localization. J Virol. 2015 Mar;89(5):2792-800. doi: 10.1128/JVI.02693-14. PMID:25540369
Note on publication:
The original publication introduces a novel method utilizing camelid-derived single-domain antibody fragments to specifically target and disrupt the function of influenza virus nucleoprotein in living cells.