Production and Purification of Zika Virus NS1 Glycoprotein in HEK293 Cells

Abstract

Zika virus (ZIKV) is an emerging mosquito-borne flavivirus which has recently caused global epidemics with its association with congenital Zika syndrome such as severe microcephaly.The recombinant ZIKV envelope (Env) glycoprotein is useful for immunological applications such as serodiagnosis of ZIKV infection and for monitoring immune responses in preclinical and clinical ZIKV vaccine developments. In this chapter, we describe the optimization of production of Zika virus envelope glycoprotein in Human Embryonic Kidney (HEK 293T) cells by small-scale expression followed by large-scale protein production. Small-scale expression of HEK 293T cells allows screening of a large number of vectors simultaneously to select the vectors with best secretory profiles for scale-up in Expi293 mammalian system to maximise the protein yield followed by purification for research and clinical applications.

Platelet activation and aggregation response to dengue virus nonstructural protein 1 and domains

“…These proteins have been successfully used for seroprevalence studies. Different from some other studies, our NS1 proteins were produced in a mammalian expression system based on HEK293, a human embryonic kidney cell line 24 . Protein expression in mammalian systems ensures post‐translational modifications such as N‐glycosilation 25 that are not present in prokaryotic produced proteins and are important to maintain the stability of dimers and hexamers secreted into circulation, as well as for their interaction with host cells.…”