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.
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.
Puttikhunt
Chunya
C
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
Ong-Ajchaowlerd
Prapapun
P
Prommool
Tanapan
T
Sangiambut
Sutha
S
Netsawang
Janjuree
J
Limjindaporn
Thawornchai
T
Malasit
Prida
P
Kasinrerk
Watchara
W
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
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