Polyclonal antibodies against recombinant dengue virus NS3 protein

Liliana Morales, Myriam L. Velandia, María Angélica Calderon, Jaime E. Castellanos, Jacqueline Chaparro-Olaya, .

DOI: https://doi.org/10.7705/biomedica.v37i1.3249
Keywords: Dengue virus, recombinant protein, immunofluorescence, immunoprecipitation, antibody production, blotting, Western
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Abstract

Introduction: Dengue is a disease caused by one of four serotypes of the dengue virus (DENV) and is endemic in approximately 130 countries. The incidence of dengue has increased dramatically in recent decades, as well as the frequency and magnitude of outbreaks. Despite all efforts, there are no prophylactic or therapeutic treatments for the disease. Accordingly, research on the processes governing the DENV infection cycle is essential to develop vaccines or antiviral therapies. One of the
most attractive DENV molecules to investigate is nonstructural protein 3 (NS3), which is essential for viral replication and a major immune target for infection.
Objective: To produce antibodies to support future studies on NS3 and its cellular interactions with other proteins.
Materials and methods: Two recombinant proteins of the helicase domain of DENV NS3 serotype 2 were expressed, and used to immunize mice and produce polyclonal antibodies.
Results: The antibodies produced were useful in Western blot and immunofluorescence tests. We report an NS3 antibody that immunoprecipitates the viral protein and detects it in Western blot with no need to over-express it or use cell extracts with metabolic radiolabeling.
Conclusion: The recombinant proteins expressed and the antibodies produced constitute valuable tools for studying DENV infectious processes involving NS3 and for evaluating tests designed to interfere with its functions.

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  • Liliana Morales Instituto de Biología Molecular, Laboratorio de Parasitología Molecular, Universidad El Bosque, Bogotá, D.C., Colombia
  • Myriam L. Velandia Instituto de Virología, Universidad El Bosque, Bogotá, D.C., Colombia
  • María Angélica Calderon Instituto de Virología, Universidad El Bosque, Bogotá, D.C., Colombia
  • Jaime E. Castellanos Instituto de Virología, Universidad El Bosque, Bogotá, D.C., Colombia
  • Jacqueline Chaparro-Olaya Instituto de Biología Molecular, Laboratorio de Parasitología Molecular, Universidad El Bosque, Bogotá, D.C., Colombia

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How to Cite
1.
Morales L, Velandia ML, Calderon MA, Castellanos JE, Chaparro-Olaya J. Polyclonal antibodies against recombinant dengue virus NS3 protein. biomedica [Internet]. 2017 Jan. 24 [cited 2024 May 18];37(1):131-40. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/3249

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Published
2017-01-24
Issue
Vol. 37 No. 1 (2017)
Section
Technical note

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