Development and Characterization of Monoclonal Antibodies Against NS1 Protein of Duck Tembusu Virus

To produce monoclonal antibodies (MAbs) against NS1 protein of duck Tembusu virus (TMUV), the NS1 gene sequence was cloned into the prokaryotic expression vector pET-28a(+). The prokaryotic protein NS1 was successfully expressed in BL21 (DE3) and used as the immunogen in mice. Six MAbs against NS1 protein were obtained by using the standard hybridoma technique. All MAbs can react with the denatured NS1 protein in the Western blot assay and the native NS1 protein from the TMUV-infected BHK-21 cells in the immunofluorescence assay. The ELISA titers of the cell supernatants and ascites of MAbs were at a high level. The subtypes of the MAbs were determined by the Rapid Mouse Isotyping Kit-Gold series. Six MAbs possessed higher specificity and sensitivity, which indicated that MAbs against NS1 protein of TMUV may be used as valuable tools for analysis of the protein functions and pathogenesis of TMUV.