1. Expression of recombinant dengue virus type 1 non-structural protein 1 in mammalian cells and preliminary assessment of its suitability to detect human IgG antibodies elicited by viral infection.
- Author
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Malnero CM, Azevedo RC, Bergmann IE, de Meneses MDF, Cavalcanti AC, Ibañez LI, and Malirat V
- Subjects
- Animals, Humans, HEK293 Cells, Sensitivity and Specificity, Antibodies, Viral, Viral Nonstructural Proteins, Enzyme-Linked Immunosorbent Assay methods, Immunoglobulin G metabolism, Mammals, Dengue Virus genetics, Dengue diagnosis, Virus Diseases
- Abstract
In recent years dengue has become a rapidly growing public health problem worldwide, however, the availability of accurate and affordable diagnostic immunoassays is limited, partly due to the difficulty of producing large quantities of purified antigen. Non-structural protein 1 (NS1) has shown to be a good candidate for inclusion in diagnostic assays and for serosurveys, particularly in endemic countries as a prerequisite for vaccination. In this work the NS1 antigen derived from dengue virus type-1 (DENV1) was expressed in HEK293-T cells and purified by affinity chromatography. The recombinant protein was recovered properly folded as dimers, highly purified and with good yield (1.5 mg/L). It was applied as a serological probe in an indirect ELISA developed in this work to detect human IgG antibodies. Preliminary comparative performance values of 81.1% sensitivity and 83.0% specificity of the developed and preliminary validated iELISA, relative to a commercial kit were obtained, suggesting that the purified recombinant DENV1 NS1 antigen is suitable to detect IgG antibodies, indicative of past DENV infection., (Copyright © 2023 Elsevier B.V. All rights reserved.)
- Published
- 2023
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