Abstract
Duck hepatitis A virus (DHAV), the major pathogen of duck virus hepatitis (DVH), causes severe diseases that threaten the duck industry worldwide. The VP1 protein, a major structural protein of DHAV, is able to induce neutralizing antibody in ducks. The purpose of this study was to identify the antigenic mimotope of DHAV by phage display technology. A monoclonal antibody (mAb) 4E6 against DHAV-1 and DHAV-3 was prepared, and a phage library prepared with the PhD-12 Phage Display Peptide Library Kit was screened with the mAb. A novel peptide, 1GLTWKLPPSM10 was identified with high affinity to the mAb and could specifically block mAb 4E6 from binding DHAV-1 and DHAV-3. Animal tests confirmed that the immunization of ducklings with the mimotope could inhibit the virus proliferation and protect the ducklings from DVH. In summary, the neutralizing conformational mimotope 1GLTWKLPPSM10 might be a promising vaccine candidate for the prevention of DHAV infection.
Highlights
Duck hepatitis A virus (DHAV) causes highly contagious and acute duck viral hepatitis (DVH) to ducklings within 3 weeks old, concomitant with liver necrosis and hemorrhage, and high mortality rate
We identified a conformational epitope, which could be effectively recognized by monoclonal antibody 4E6 of DHAV through a 12-mer random peptide phage display library
4E6 neutralized DHAV-1 LY0801 strain with a neutralization titer of 42.2 dilution fold, whereas it neutralized the DHAV-3 SD1201 strain with a neutralization titer of 22.6 dilution fold, which indicated that DHAV-1 was the predominant serotype recognized by monoclonal antibody (mAb) 4E6
Summary
Duck hepatitis A virus (DHAV) causes highly contagious and acute duck viral hepatitis (DVH) to ducklings within 3 weeks old, concomitant with liver necrosis and hemorrhage, and high mortality rate. As one of the pathogens that seriously jeopardize duck industry in Southeast Asia, an outbreak of DHAV infection will usually be followed by great economy losses [1,2]. DHAV has been classified into three serotypes according to results of neutralization tests, namely the classical serotype 1. The mixed infections of different DHAVs had happened with increasing frequency in eastern Asia [1,2,6], intensifying the difficulty of DVH prevention and control. Excluding the poly(A) tail, which is irregular in length, the single-stranded positive-sense RNA of the complete DHAV genome is approximately 7700 nucleotides. It is encapsulated in an icosahedral structure, which is assembled
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