Abstract
Dengue is considered a reemerging disease of worldwide distribution. The Dengue virus non-structural protein 3 (NS3) is known to possess ATPase, helicase, and protease activities that are a constitutive part of the replication complex of Dengue virus. In this report, we discuss the cloning, expressing, and purifying of the Dengue-2 NS3 protein, to immunize mice and then generate monoclonal antibodies (MAbs). Our results show the production of MAbs specific to NS3 protein of Dengue-2 virus, which by immunofluorescence recognize the native protein in experimentally infected endothelial cells (HMEC). Likewise, C6/36-infected lisates were used in Western blots, and observed the specific characteristic band that defines the NS3 protein. We conclude that these antibodies may be a useful tool, not only to study the replicative process of Dengue virus, but also to generate specific diagnostic tools for Dengue infection.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphatases / immunology*
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / immunology*
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Antibody Specificity / immunology*
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Blotting, Western
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Cloning, Molecular
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Dengue Virus / chemistry
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Dengue Virus / immunology*
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Dengue Virus / isolation & purification
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Electrophoresis, Polyacrylamide Gel
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Endothelial Cells / virology
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli / genetics
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Fluorescent Antibody Technique, Indirect
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Glutathione Transferase / metabolism
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Humans
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Immunization*
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Mice
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Mice, Inbred BALB C
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Microscopy, Fluorescence
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Polymerase Chain Reaction
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Recombinant Fusion Proteins / immunology
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Recombinant Fusion Proteins / metabolism
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Serine Endopeptidases / genetics
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Serine Endopeptidases / immunology*
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Serine Endopeptidases / isolation & purification
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Serine Endopeptidases / metabolism
Substances
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Antibodies, Monoclonal
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Recombinant Fusion Proteins
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Glutathione Transferase
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NS3 protease, dengue virus
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Serine Endopeptidases
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Adenosine Triphosphatases