Development of a chicken interferon-induced transmembrane protein 3 (IFITM3)-specific monoclonal antibody using phage display.

Abstract

Interferon-induced transmembrane protein 3 (IFITM3) has potent antiviral activity against several viruses. Recent studies have reported that the chicken IFITM3 gene also plays a pivotal role in blocking viral replication, but these studies are considerably limited due to being conducted at the RNA level only. Thus, the development of a chicken IFITM3 protein-specific antibody is needed to validate the function of IFITM3 at the protein level. Epitope prediction was performed with the immune epitope database analysis resource (IEDB-AR) program. The epitope was validated by four in silico programs, Jped4, Clustal Omega, TMpred and SOSUI. Chicken IFITM3 protein-specific monoclonal antibodies were screened by enzyme-linked immunosorbent assay through affinity between recombinant IFITM3 protein and phage-displayed candidate antibodies. Validation of the reactivity of the chicken IFITM3 protein-specific antibody to chicken tissues was carried out using western blotting. We developed a chicken IFITM3 protein-specific monoclonal antibody using phage display. The reactivity of the antibody with peripheral chicken tissues was confirmed using western blotting. To the best of our knowledge, this was the first development of a chicken IFITM3 protein-specific monoclonal antibody using phage display.