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Characterization and Application of a New Monoclonal Antibody with High Specificity for Helicobacter hepaticus |
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| Authors: | Yoshihiro Fukuda Tadashi Shimoyama Takeo Ohmura Yukari Sano Natsuko Nakabayashi Ryoki Takahashi Toshio Fujioka Hazel M Mitchell Takashi Shimoyama |
| Institution: | Division of Clinical Nutrition, Hyōgo College of Medicine, Nishinomiya, Hyōgo, Japan,;Department of Gastroenterology, Hirosaki University School of Medicine, Hirosaki, Aomori, Japan,;Gastrointestinal Research Sagami Research Laboratories, Wakamoto Pharmaceutical Co., Ltd., Kanagawa, Japan,;Department of Gastroenterology, Ōita University, School of Medicine, Yufu, Ōita, Japan,;School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales, Australia |
| Abstract: | Background and Aims: Infection with Helicobacter hepaticus is suggested to play a role in the pathogenesis of chronic liver disease in humans. However, reactive antigens among Helicobacter species make the development of an H. hepaticus ELISA test with high specificity difficult. A new monoclonal antibody from a hybridoma clone (HRII-51) showed high specificity to H. hepaticus without cross-reaction to other gastrointestinal bacteria. Methods: The molecular weight of HRII-51 immunoreactive antigen was examined by Western blot of H. hepaticus probed with the monoclonal antibody HRII-51. A HRII-51-immunoreactive antigen capture ELISA was prepared in which the specific antigen was anchored by HRII-51-immobilized ELISA plate. Accuracy of HRII-51 antigen capture ELISA was examined using sera obtained from mice inoculated with Helicobacter species. Specificity of HRII-51 antigen capture ELISA was compared to that of H. hepaticus antigen-based ELISA using human sera with absorption by H. pylori cell lysate. Results: HRII-51 immunoreactive antigen had a molecular weight of 15 kDa. Sensitivity and specificity of HRII-51 antigen capture ELISA were 87.0% and 97.6% in mice inoculated with Helicobacter species. In human sera, modification of the results by absorption with H. pylori lysate was smaller in HRII-51 antigen capture ELISA comparing with H. hepaticus -antigen-based ELISA. Conclusion: Use of the HRII-51 antigen capture ELISA would be a useful approach for the serodiagnosis of H. hepaticus infection in both experimental animals and humans. |
| Keywords: | Helicobacter hepaticus ELISA monoclonal antibody |