Production of a monoclonal antibody against non-structural protein 3 of dengue-2 virus by intrasplenic injection.

Dengue fever and dengue hemorrhagic fever/dengue shock syndrome are highly infectious diseases caused by dengue virus (DV). DV non-structural protein 3 (NS3) is known to possess ATPase, helicase, and protease activity that is a constitutive part of the replication complex of DV. In this study, we discuss the cloning, expression, and purification of the DV-2 NS3 protein to immunize mice by intrasplenic injection and then to generate a monoclonal antibody (MAb). One MAb, named 4F5, was obtained and it was specific to NS3 of DV-2. Immunofluorescence show that 4F5 recognizes the native protein in infected ECV304 cells. Likewise, C6/36-infected lysates were used in Western blot analysis, and we observed the specific characteristic band that defines NS3. We conclude that MAb 4F5 may be a useful tool, not only to study the replicative process of DV, but also to generate specific diagnostic tools for DV infection.