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Generation and Characterization of Monoclonal Antibodies to the Ogawa Lipopolysaccharide of Vibrio cholerae O1 from Phage-Displayed Human Synthetic Fab Library

Journal of Microbiology and Biotechnology 2020년 30권 11호 p.1760 ~ 1768
김다인, 홍지수, 최윤주, 한제민, 김상규, 조경희, 윤준열, 채희수, 윤혜선, 이찬규, 홍효정,
소속 상세정보
김다인 ( Kim Da-In ) - Kangwon National University College of Biomedical Science Department of Systems Immunology
홍지수 ( Hong Ji-Su ) - Kangwon National University College of Biomedical Science Department of Systems Immunology
최윤주 ( Choi Yoon-Joo ) - Chonnam National University Medical School Medical Research Center
한제민 ( Han Je-Min ) - Eubiologics Co., Ltd.
김상규 ( Kim Sang-Kyu ) - Kangwon National University College of Biomedical Science Department of Systems Immunology
조경희 ( Jo Gyung-Hee ) - Kangwon National University College of Biomedical Science Department of Systems Immunology
윤준열 ( Yoon Jun-Yeol ) - Kangwon National University College of Biomedical Science Department of Systems Immunology
채희수 ( Chae Hee-Su ) - Kangwon National University College of Biomedical Science Department of Systems Immunology
윤혜선 ( Yoon Hye-Seon ) - Eubiologics Co., Ltd.
이찬규 ( Lee Chan-Kyu ) - Eubiologics Co., Ltd.
홍효정 ( Hong Hyo-Jeong ) - Kangwon National University College of Biomedical Science Department of Systems Immunology

Abstract


Vibrio cholerae, cause of the life-threatening diarrheal disease cholera, can be divided into different serogroups based on the structure of its lipopolysaccharide (LPS), which consists of lipid-A, corepolysaccharide and O-antigen polysaccharide (O-PS). The O1 serogroup, the predominant cause of cholera, includes two major serotypes, Inaba and Ogawa. These serotypes are differentiated by the presence of a single 2-O-methyl group in the upstream terminal perosamine of the Ogawa O-PS, which is absent in the Inaba O-PS. To ensure the consistent quality and efficacy of the current cholera vaccines, accurate measurement and characterization of each of these two serotypes is highly important. In this study, we efficiently screened a phage-displayed human synthetic Fab library by bio-panning against Ogawa LPS and finally selected three unique mAbs (D9, E11, and F7) that specifically react with Ogawa LPS. The mAbs bound to Vibrio cholerae vaccine in a dose-dependent fashion. Sequence and structure analyses of antibody paratopes suggest that IgG D9 might have the same fine specificity as that of the murine mAbs, which were shown to bind to the upstream terminal perosamine of Ogawa O-PS, whereas IgGs F7 and E11 showed some different characteristics in the paratopes. To our knowledge, this study is the first to demonstrate the generation of Ogawa-specific mAbs using phage display technology. The mAbs will be useful for identification and quantification of Ogawa LPS in multivalent V. cholerae vaccines.

키워드

Vibrio cholerae; Ogawa serotype; O-antigen polysaccharide; monoclonal antibody; phage display

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