title: Development of a monoclonal antibody specific to envelope domain III with broad-spectrum detection of all four dengue virus serotypes.
abstract: Dengue virus (DENV) is a mosquito-borne pathogen that annually infects more than 390 million people in 100 different countries. Symptoms of the viral infection include a relatively weak dengue fever to severe dengue hemorrhagic fever/dengue shock syndrome, which are mortal infectious diseases. As of yet, there is no commercially available vaccine or therapeutic for DENV. Currently, passive immunotherapy using DENV-specific antibody (Ab) is a considered strategy to treat DENV infection. Here, we developed a monoclonal Ab (mAb), EDIIImAb-61, specific to the DENV domain III of the envelope glycoprotein (EDIII) with broad-spectrum detection ability to all four DENV serotypes (DENV-1∼4) to use as a therapeutic Ab. Although EDIII contains non-immunodominant epitopes compared to domains I and II, domain III plays a critical role in host receptor binding. EDIIImAb-61 exhibited cross-reactive binding affinity to all four DENV serotypes that had been isolated from infected humans. To further characterize EDIIImAb-61 and prepare genes for large-scale production using a heterologous expression system, the sequence of the complementarity determining regions was analyzed after cloning the full-length cDNA genes encoding the heavy and light chain of the mAb. Finally, we produced Ab from CHO-K1 cells transfected with the cloned EDIIImAb-61 heavy and light chain genes and confirmed the binding ability of the Ab. Collectively, we conclude that EDIIImAb-61 itself and the recombinant Ab produced using the cloned heavy and light chain gene of EDIIImAb-61 is a candidate for passive immunotherapy against DENV infection.
authors.affiliation: Department of Molecular Biology and the Institute for Molecular Biology and Genetics, Chonbuk National University, Jeonju 54896, South Korea; Department of Bioactive Material Sciences and Research Center of Bioactive Materials, Jeonju 54896, South Korea.
authors.forename: Sae-Hae
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Department of Bioactive Material Sciences and Research Center of Bioactive Materials, Jeonju 54896, South Korea.
authors.forename: Yu Na
authors.initials: YN
authors.lastname: Kim
authors.affiliation: Canadian Food Inspection Agency, National Center for Foreign Animal Disease, Winnipeg, MB R3E 3M4, Canada.
authors.forename: Thang Thua
authors.initials: TT
authors.lastname: Truong
authors.affiliation: National Institute of Hygiene and Epidemiology, Hanoi, Viet Nam.
authors.forename: Nguyen Thi
authors.initials: NT
authors.lastname: Thu Thuy
authors.affiliation: National Institute of Hygiene and Epidemiology, Hanoi, Viet Nam.
authors.forename: Le Quynh
authors.initials: LQ
authors.lastname: Mai
authors.affiliation: Department of Molecular Biology and the Institute for Molecular Biology and Genetics, Chonbuk National University, Jeonju 54896, South Korea; Department of Bioactive Material Sciences and Research Center of Bioactive Materials, Jeonju 54896, South Korea. Electronic address: yongsuk@jbnu.ac.kr.
authors.forename: Yong-Suk
authors.initials: YS
authors.lastname: Jang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology*; D001665:Binding Sites; D016466:CHO Cells; D003412:Cricetulus; D003716:Dengue Virus / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D015195:Drug Design; D011485:Protein Binding; D015202:Protein Engineering; D065288:Serogroup; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D014759:Viral Envelope Proteins
keywords: Dengue virus; Envelope glycoprotein domain III; Monoclonal antibody; Passive immunotherapy