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

J Immunol Methods. 2023 Jul:518:113503. doi: 10.1016/j.jim.2023.113503. Epub 2023 May 30.

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.

Keywords: Dengue virus; Diagnosis; Mammalian expressed antigen; Non-structural protein 1; Pre-vaccination screening; Seroprevalence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Viral
  • Dengue Virus* / genetics
  • Dengue* / diagnosis
  • Enzyme-Linked Immunosorbent Assay / methods
  • HEK293 Cells
  • Humans
  • Immunoglobulin G / metabolism
  • Mammals
  • Sensitivity and Specificity
  • Viral Nonstructural Proteins
  • Virus Diseases*

Substances

  • Antibodies, Viral
  • Viral Nonstructural Proteins
  • Immunoglobulin G