Among them, NS1 protein is a glycoprotein which is present in cell-surface or in the form of cell-associated protein . minimal determinant of the linear B cell epitope was recognized and identified with mAb 3G2. The accurate linear B-cell epitope was 269DEKEIV274 located in NS1 protein. Furthermore, sequence alignment showed that the epitope was highly conserved and specific among TMUV strains and other flavivirus respectively. The linear B-cell epitope of TMUV NS1 protein could benefit the development of new vaccines and diagnostic assays. Introduction Tembusu virus infection in ducks is caused by Tembusu virus (TMUV), which was first reported in April 2010 in AZD4547 China. The virus can infect more varieties of ducks such as Beijing duck, golden duck, Shaoxing duck, Cherry Valley, Campbell ducks, Jinyun duck, etc. Laying ducks infected mainly demonstrated drops in egg production, follicular rupture and bleeding, and yolk peritonitis. Ducklings mainly displayed standing instability and paralysis, retarded growth, with 10 to 30% mortality rates [2, 3]. Other birds such as Chickens, geese, sparrows, etc were also infected and displayed obvious clinical signs [4C6]. So far, the disease has resulted in a great economic loss to the poultry industry and caused wide public concern. There is no specific treatment available for TMUV and the AZD4547 vaccination is an effective way to prevent TMUV infection in waterfowl. The inactivated vaccines and live attenuated vaccines against TMUV have been successfully developed and already used in clinical production[7, 8]. But the live attenuated vaccines could display disadvantages of reversion of virulence and spread, and the inactivated vaccines didnt display an effective cellular immunity. So the development of a new type of vaccine is very urgent. TMUV is a mosquito-borne flavivirus which belongs to the Ntaya virus group within flavivirus genus, flaviviridae family . TMUV genome encodes a single polyprotein which is cleaved into three structural proteins (C, prM and E) and seven non-structural proteins (NS1, NS2A, NS2B, NS3, NS4A, NS4B and NS5) . Among them, NS1 protein is a glycoprotein which is present Rabbit Polyclonal to Cyclin L1 in cell-surface or in the form of cell-associated protein . NS1 protein possesses several unusual and interesting performances which is closely related AZD4547 to the membrane function and indispensable in the early viral replication, assembly and release of the virus . NS1 protein contains multiple protective T-cell and B-cell epitopes which can induce both humoral and cell-mediated immunity without the risk of antibody-dependent enhancement [12, 13]. The B-cell epitopes mainly refer to multiple continuous amino acid residues on the protein surface or the spatial conformation of discontinuous amino acid residues. Accurate analysis of the epitopes on NS1 protein is critical for further understanding the mechanism of NS1-mediated immune protection. In recent years, epitope-based marker vaccines have increasingly attracted wide attentions in public . The identification of linear epitopes on NS1 protein would be conducive to the development of epitope-based marker and subunit vaccines, preparation of protective antibodies and understanding protein functions . It has been demonstrated that NS1 protein AZD4547 of flavivirus has more virus-specific epitopes than cross-reactive types as opposed to E proteins which has even more cross-reactive than particular epitopes . As a result, it’s very interesting and precious to display screen and recognize epitopes on NS1 proteins for particular serological diagnosis that may differentiate between attacks due to flaviviruses [16C18]. In this scholarly study, one linear B-cell epitope was characterized and identified with one monoclonal antibody against TMUV NS1 proteins. This scholarly research can place the building blocks for comprehending the antigenic framework of TMUV NS1 proteins, advancement of a particular serological diagnostic assay and a fresh kind of vaccine for TMUV an infection. Materials and strategies Ethics declaration This analysis was accepted by the Committee over the Ethics of Pet of Shandong (permit amount 20147620). Cell lines Myeloma cell series SP2/0 and baby hamster kidney BHK-21 cells (CCL-10, American Type Lifestyle Collection) had been cultured in DMEM/Great blood sugar (Hyclone, Thermo technological, USA) within a humidified 5% CO2 atmosphere at 37C. All lifestyle media had been added with 10% inactivated fetal bovine serum (TransGen, Beijing, China), 100 U/mL penicillin and 100 mg/mL streptomycin (Solarbio, Beijing, China). Serum and Virus.