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Open AccessArticle

NS1 Recombinant Proteins Are Efficiently Produced in Pichia pastoris and Have Great Potential for Use in Diagnostic Kits for Dengue Virus Infections

1
Department of General Biology, Federal University of Viçosa, Viçosa, Minas Gerais 36570-900, Brazil
2
Department of Microbiology, Federal University of Viçosa, Viçosa, Minas Gerais 36570-900, Brazil
*
Author to whom correspondence should be addressed.
Diagnostics 2020, 10(6), 379; https://doi.org/10.3390/diagnostics10060379
Received: 20 April 2020 / Revised: 26 May 2020 / Accepted: 4 June 2020 / Published: 6 June 2020
(This article belongs to the Section Point-of-Care Diagnostics and Devices)
Dengue is one of the major diseases causing global public health concerns. Despite technological advances in vaccine production against all its serotypes, it is estimated that the dengue virus is responsible for approximately 390 million infections per year. Laboratory diagnosis has been the key point for the correct treatment and prevention of this disease. Currently, the limiting factor in the manufacture of dengue diagnostic kits is the large-scale production of the non-structural 1 (NS1) antigen used in the capture of the antibody present in the infected patients’ serum. In this work, we demonstrate the production of the non-structural 1 protein of dengue virus (DENV) serotypes 1–4 (NS1-DENV1, NS1-DENV2, NS1-DENV3, and NS1-DENV4) in the methylotrophic yeast Pichia pastoris KM71H. Secreted recombinant protein was purified by affinity chromatography and characterized by SDS-PAGE and ELISA. The objectives of this study were achieved, and the results showed that P. pastoris is a good heterologous host and worked well in the production of NS1DENV 1–4 recombinant proteins. Easy to grow and quick to obtain, this yeast secreted ready-to-use proteins, with a final yield estimated at 2.8–4.6 milligrams per liter of culture. We reached 85–91% sensitivity and 91–93% specificity using IgM as a target, and for anti-dengue IgG, 83–87% sensitivity and 81–93% specificity were achieved. In this work, we conclude that the NS1 recombinant proteins are efficiently produced in P. pastoris and have great potential for use in diagnostic kits for dengue virus infections. The transformed yeast obtained can be used for production in industrial-scale bioreactors. View Full-Text
Keywords: dengue; NS1 protein; Pichia pastoris; diagnosis dengue; NS1 protein; Pichia pastoris; diagnosis
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MDPI and ACS Style

Xisto, M.F.; Prates, J.W.O.; Dias, I.M.; Dias, R.S.; da Silva, C.C.; de Paula, S.O. NS1 Recombinant Proteins Are Efficiently Produced in Pichia pastoris and Have Great Potential for Use in Diagnostic Kits for Dengue Virus Infections. Diagnostics 2020, 10, 379.

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