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Generation and Characterization of Antinonstructural Protein 1 Monoclonal Antibodies and Development of Diagnostics for Dengue Virus Serotype 2

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  • 1 Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan;
  • | 2 Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan;
  • | 3 Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung-Hsing University, Taichung, Taiwan;
  • | 4 Institute of Epidemiology and Preventive Medicine, College of Public Health, National Taiwan University, Taipei, Taiwan;
  • | 5 Division of Infectious Diseases, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan;
  • | 6 School of Medicine, Graduate Institute of Medicine, Sepsis Research Center, Kaohsiung Medical University, Kaohsiung, Taiwan;
  • | 7 Department of Biological Science and Technology, College of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan

Dengue virus (DENV) circulates in tropical and subtropical areas around the world, where it causes high morbidity and mortality. There is no effective treatment of infection, with supportive care being the only option. Furthermore, early detection and diagnosis are important to facilitate clinical decisions. In this study, seven monoclonal antibodies (mAbs) recognizing nonstructural protein 1 (NS1) of DENV were generated by hybridoma techniques. These antibodies can be divided into two groups: serotype-specific (DB6-1, DB12-3, and DB38-1) and nonspecific (consisting of antibodies DB16-1, DB20-6, DB29-1, and DB41-2). The B-cell epitopes of DB20-6 and DB29-1 were identified by phage display and site-directed mutagenesis, and its binding motif, WXXWGK, was revealed to correspond to amino acid residues 115–120 of the DENV-2 NS1 protein. A diagnostic platform, consisting of a serotype-specific capture antibody and a complex detection antibody, exhibited a detection limit of about 1 ng/mL, which is sufficient to detect NS1 in clinical serum samples from dengue patients. This diagnostic platform displayed better specificity and sensitivity than two examined commercial NS1 diagnostic platforms. In summary, our results indicate that these newly generated mAbs are suitable for detection of NS1 protein of DENV-2 in clinical samples.

Author Notes

Address correspondence to Han-Chung Wu, Institute of Cellular and Organismic Biology, Academia Sinica, 128 Academia Road, Section 2, Nankang, Taipei 11529, Taiwan. E-mail: hcw0928@gate.sinica.edu.tw

Financial support: This research was supported by grants from Academia Sinica and the Ministry of Science and Technology (104-0210-01-09-02), Taiwan (to H-C Wu).

Authors’ addresses: Yin-Liang Tang, I-Ju Liu, Pi-Chun Li, Chien-Yu Chiu, Chi-Yu Fu, and Han-Chung Wu, Institute of Cellular and Organismic Biology, Academia Sinica, Taipei, Taiwan, E-mails: bigerliang@yahoo.com.tw, spinner.striped@msa.hinet.net, pichunli1210@yahoo.com.tw, chienyu412@gmail.com, fuchiyu@gate.sinica.edu.tw, and hcw0928@gate. sinica. edu.tw. Chun-Yu Lin, Chung-Hao Huang, and Yen-Hsu Chen, Department of Internal Medicine, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan, E-mails: infectionman@gmail.com, locusthao@gmail.com, and infchen@gmail.com. Day-Yu Chao, Graduate Institute of Microbiology and Public Health, College of Veterinary Medicine, National Chung-Hsing University, Taichung, Taiwan, E-mail: dychao@dragon.nchu.edu.tw. Chwan-Chuen King, Institute of Epidemiology and Preventive Medicine, College of Public Health, National Taiwan University, Taipei, Taiwan, E-mail: chwanchuen@gmail.com.

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