Expression and purification of NS1 protein for designing a lateral flow assay-based aptamer-antibody

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Expression and purification of NS1 protein for designing a lateral flow assay-based aptamer-antibody

Scientific Reports , Article number:  (2025) Cite this article

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Abstract

Dengue fever, a mosquito-borne viral disease, presents with mild symptoms in most patients but is characterized by dengue hemorrhagic fever (DHF) in severe cases. Current diagnostic methods are expensive and take time to process. Diagnostic testing is currently limited, particularly in low-resource environments. In this work, we provide the development of a rapid diagnostic test for dengue using a lateral flow immunoassay (LFA) strip using an aptamer-antibody sandwich to detect the NS1 protein. The LFA includes an antibody capture zone and a conjugate pad with an aptamer linked to gold nanoparticles (AuNPs). A biotin-labeled complementary aptamer is immobilized in the control zone on streptavidin, forming a sandwich format. In the presence of the NS1 protein, it binds to the antibody and the aptamer-AuNP complex, producing a visible bichromatic line to indicate a positive result. The LFA detects DENV2 NS1 at concentrations as low as 5.2 ng/mL (95% CI: 4.6–5.8) in buffer. With a detection limit of 6.1 ng/mL (95% CI: 5.5–6.8) in spiked human serum, the test may offer a promising approach, pending further clinical evaluation approach for early dengue diagnosis.

Data availability

Yes, data is provided within the manuscript file.

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Acknowledgements

This work was supported by the University of Tehran and Arvin BioHealth Company, Tehran, Iran. We want to thank Mohammad Hossein Khodabandeloo from the Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran, for the assistance in drawing our shapes.

Author information

Authors and Affiliations

  1. Department of Technical and Research Lab, Arvin BioHealth Company, Tehran, Iran

    Mohammad Javad Jadidi & Hassan Morovati Khamsi

  2. Institute of Biochemistry and Biophysics (IBB), University of Tehran, Tehran, Iran

    Mohammad Javad Jadidi, Ahmad Reza Ghaffari, Amir Hossein Tohidi & Majid Mahdavi

  3. Graduate Research Assistant, Department of Cell and Molecular Biology and Microbiology, University of Isfahan, Isfahan, Iran

    Hamidreza Majidifard

  4. Department of Quality Control, Razi Vaccine and Serum Research Institute, Agricultural Research Education and Extension Organization, Karaj, Iran

    Hassan Morovati Khamsi

Authors

  1. Mohammad Javad Jadidi
  2. Hamidreza Majidifard
  3. Ahmad Reza Ghaffari
  4. Amir Hossein Tohidi
  5. Hassan Morovati Khamsi
  6. Majid Mahdavi

Contributions

MJ J: Conceptualization, methodology, formal analysis, investigation, resources, and original draft preparation. HR M: Review & Editing. AH T: Methodology, writing original draft preparation. AR G: Methodology, writing original draft preparation. H M-K: Conceptualization, methodology, formal analysis, validation, data curation, supervision. M M: Conceptualization, methodology, formal analysis, validation investigation, review and editing, and supervision.

Corresponding authors

Correspondence to Hassan Morovati Khamsi or Majid Mahdavi.

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The authors declare no competing interests.

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Jadidi, M.J., Majidifard, H., Ghaffari, A. et al. Expression and purification of NS1 protein for designing a lateral flow assay-based aptamer-antibody. Sci Rep (2025). https://doi.org/10.1038/s41598-025-29869-4

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  • DOI: https://doi.org/10.1038/s41598-025-29869-4

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