Abstract
We produced monoclonal and polyclonal antibodies to the capsid (C) protein of dengue serotype 2 virus (DV2 C). First, a maltose-binding protein fused to DV2 C protein (MBP-C) was overproduced in E. coli. The affinity-purified MBP-C protein was cleaved by factor Xa protease to obtain a recombinant DV2 C protein, which was then used for mouse immunizations. Two hybridoma cell lines producing anti-C Mabs as well as anti-C polyclonal antibody were successfully generated and characterized. Interestingly, all of the generated antibodies specifically recognized the first 20 amino acids of the DV2 C protein, as determined by peptide epitope mapping and via a recombinant DV2 C protein in which this region was deleted. The results suggested that this region is predominantly immunogenic in mice.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Animals
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Antibodies, Monoclonal / biosynthesis
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Antibodies, Monoclonal / immunology*
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Antibodies, Viral / biosynthesis
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Antibodies, Viral / immunology*
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Capsid Proteins / genetics
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Capsid Proteins / immunology*
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Capsid Proteins / metabolism
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Carrier Proteins / metabolism
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Dengue / immunology*
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Dengue Virus / immunology*
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Epitope Mapping
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Factor Xa / metabolism
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Immunization
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Immunodominant Epitopes / genetics
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Immunodominant Epitopes / immunology*
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Maltose-Binding Proteins
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Mice
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Mice, Inbred BALB C
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Molecular Sequence Data
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Peptide Hydrolases / metabolism
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Recombinant Proteins / immunology
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Recombinant Proteins / metabolism
Substances
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Antibodies, Monoclonal
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Antibodies, Viral
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Capsid Proteins
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Carrier Proteins
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Immunodominant Epitopes
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Maltose-Binding Proteins
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Recombinant Proteins
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Peptide Hydrolases
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Factor Xa