title: Production and characterization of anti-dengue capsid antibodies suggesting the N terminus region covering the first 20 amino acids of dengue virus capsid protein is predominantly immunogenic in mice.
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.
authors: Medical Biotechnology Unit, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 12th Floor Adulyadej-Vikrom Building, Siriraj Hospital, Bangkok 10700, Thailand. chunyapk@biotec.or.th
authors: Chunya
authors: C
authors: Puttikhunt
authors: Prapapun
authors: P
authors: Ong-Ajchaowlerd
authors: Tanapan
authors: T
authors: Prommool
authors: Sutha
authors: S
authors: Sangiambut
authors: Janjuree
authors: J
authors: Netsawang
authors: Thawornchai
authors: T
authors: Limjindaporn
authors: Prida
authors: P
authors: Malasit
authors: Watchara
authors: W
authors: Kasinrerk
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism; D003715:Dengue / Q000276:immunology*; D003716:Dengue Virus / Q000276:immunology*; D018604:Epitope Mapping; D015951:Factor Xa / Q000378:metabolism; D007114:Immunization; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology*; D058619:Maltose-Binding Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D036022:Capsid Proteins; D002352:Carrier Proteins; D016056:Immunodominant Epitopes; D058619:Maltose-Binding Proteins; D011994:Recombinant Proteins; D010447:Peptide Hydrolases; D015951:Factor Xa