Expression and Purification of Dengue NS1 Protein in Sf9-Baculovirus System

It is well known that glycosylations of Dengue NS1 protein are important for its structure, oligomerization, and immunogenicity. One of the major challenges in heterologous NS1 protein expression is the difference in glycosylation patterns amongst different organisms. The two major natural hosts for Dengue virus are humans and mosquitoes, which are capable of producing very complex glycosylation motifs. This chapter presents an optimized protocol for heterologous expression and purification of Dengue NS1 protein in Sf9 cells infected with baculovirus. NS1 protein obtained from this protocol is glycosylated and capable of forming soluble hexamers that can be used for structural and functional assays.