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Roldán, Julieta S.; Cassola, Alejandro; Castillo, Daniela S.
Biotechnology reports (Amsterdam, Netherlands), 03/2020, Volume: 25Journal Article
•Stable recombinant ZIKV NS1-His-expressing HEK293 cells were generated.•Rapamycin treatment followed by serum starvation leads to a 29-fold increase in recombinant ZIKV NS1 protein secretion.•The purified recombinant ZIKV NS1 hexamer is a reliable biological tool for clinical diagnosis and surveillance purposes. Sensitive, accurate and cost-effective diagnostic tests are urgently needed to detect Zika virus (ZIKV) infection. Nonstructural 1 (NS1) glycoprotein is an excellent diagnostic marker since it is released in a hexameric conformation from infected cells into the patient's bloodstream early in the course of the infection. We established a stable rZNS1-His-expression system in HEK293 cells through lentiviral transduction. A novel optimization approach to enhance rZNS1-His protein secretion in the mammalian expression system was accomplished through 50 nM rapamycin incubation followed by serum-free media incubation for 9 days, reaching protein yields of ∼10 mg/l of culture medium. Purified rZNS1-His hexamer was recognized by anti-NS1 antibodies in ZIKV patient's serum, and showed the ability to induce a humoral response in immunized mice. The obtained recombinant protein is a reliable biological tool that can potentially be applied in the development of diagnostic tests to detect ZIKV in infected patients during the acute phase.
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