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Clinical and Vaccine Immunology, April 2007, p. 404-411, Vol. 14, No. 4
1071-412X/07/$08.00+0     doi:10.1128/CVI.00249-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Generation and Characterization of Monoclonal Antibodies against Dengue Virus Type 1 for Epitope Mapping and Serological Detection by Epitope-Based Peptide Antigens

Institute of Cellular and Organismic Biology, Academia Sinica, Taipei 115, Taiwan,¹ Institute of Pathology, College of Medicine, National Taiwan University, Taipei 100, Taiwan,² Institute of Epidemiology, College of Public Health, National Taiwan University, Taipei 100, Taiwan³

Received 27 June 2006/ Returned for modification 14 September 2006/ Accepted 18 January 2007

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.

Published ahead of print on 7 February 2007.

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