Generation and characterization of monoclonal antibodies against dengue virus type 1 for epitope mapping and serological detection by epitope-based peptide antigens.
Dengue virus (DEN), the pathogen behind dengue hemorrhagic fever, remains a public health problem in Asia and South America. In this study, monoclonal antibodies (MAbs) against DEN serotype 1 (DEN-1) were generated by fusing NSI/1-Ag4-1 mouse myeloma cells with lymphocytes from BALB/c mice immunized with DEN-1. Twelve MAbs were found to react specifically to the DENs by enzyme-linked immunosorbent assay, immunofluorescence analysis, and immunoblotting analysis. Five MAbs, namely, DA4-7, DA6-7, DA9-5, DA10-2, and DA11-13, were found to react with envelope proteins of DEN-1. Two serotype-specific MAbs of DEN-1, DA6-7 and DA11-13, were further shown to neutralize DEN-1 infection by a plaque reduction neutralization test. The neutralizing epitopes of these MAbs were further identified from a random peptide library displayed on phage. Immunopositive phage clones reacted specifically with these MAbs and did not react with normal mouse serum. Epitope-based peptide antigens were proved able to detect antibodies in serum samples collected from DEN-1-infected patients but not in those taken from DEN-2-infected patients or healthy controls. We believe that these MAbs and neutralizing epitopes will provide information that will lead to the development of DEN-1 serotype-specific diagnostic reagents and vaccines.
Chen
Yun-Ching
YC
Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan.
Huang
Hsien-Neng
HN
Lin
Chin-Tarng
CT
Chen
Yi-Fang
YF
King
Chwan-Chuen
CC
Wu
Han-Chung
HC
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000097:blood / Q000737:chemistry; D000956:Antigens, Viral / Q000097:blood / Q000276:immunology*; D002460:Cell Line; D003716:Dengue Virus / Q000276:immunology*; D018604:Epitope Mapping; D006801:Humans; D051379:Mice; D010455:Peptides / Q000097:blood / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D010455:Peptides