12th International Conference of Pharmaceutical Sciences

Dengue is an important public health problem worldwide, caused by four antigenically distinct dengue virus serotypes. Recent estimates indicate that there are ~390 million dengue infections per year around the world, of which ~25% result in clinical disease¹. This work describes a method for the expression of NS1 and E3 proteins in Escherichia coli, which could potentially be used to develop a point of care diagnostics kit. Synthetic full-length NS1 and E3 genes of dengue virus serotype 1 were cloned into pET28a to construct plasmid pET28a_DENVNS1 and pET28a_DENVE3, respectively. The proteins were expressed in inclusion bodies which were solubilized under denaturing conditions and the recovered proteins were purified by nickel affinity chromatography. The purified proteins were identified by Western Blotting using anti-His tag antibody. In addition, rDENVNS1 was recognized in a rapid test for detection of dengue NS1 antigens, which gives it specificity and great diagnostic potential as capture antigen.