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학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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Porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae is a highly contagious disease that leads to enormous economic losses in pig industry, worldwide. Of the many virulence factors produced by the causative bacterium, ApxA exotoxins have been considered as the most important contributor to the disease. The toxins are classified into four different types; ApxIA, ApxIIA, ApxIIIA and ApxIVA. Uniquely, ApxIVA is expressed across all serotypes of A. pleuropneumoniae only during in vivo infection in pigs. Active research focusing on resolving the precious roles and mechanisms of the toxins is still at its primitive stage. In this study, we report the development of monoclonal antibodies against the two major antigenic epitopes that were characterized in our previous study incorporating the in silico predictions and protein modeling analyses. Recombinant proteins of the selected epitopes were expressed and purified after molecular cloning of the corresponding partial genes in E. coli expression system. Subsequently, we generated hybridomas with lymphoid cells from the rats immunized with the recombinantly expressed proteins of Apx. Consequently, hybridomas exhibiting strong productivity of the monoclonal antibodies were selected for downstream verifications that tested for reactivity and specificity using Western blot and ELISA. Our results strongly suggest the potential application of the monoclonal antibodies developed in this study as useful reagents to further elaborate the mechanism of the A. pleuropneumoniae infection in pig.
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참고문헌 (15)
참고문헌 신청
Bosse JT / 2002 / Actionbacillus pleuropneumoniae : pathobiology and pathogenesis of infection / Microbes Infect 4:225~235
Huang H / 2006 / Generation of monoclonal antibodies and epitope mapping of ApxIVA of Actinobacillus pleuropneumoniae / Mol Immunol 43:2130~2134
Inzana TJ / 2011 / Serologic detection of Actinobacillus pleuropneumoniae in swine by capsular polysaccharide-biotin-streptavidin enzyme-linked immunosorbent assay / J Clin Microbiol 39:1279~1282
Jung M / 2019 / Development of Actinobacillus pleuropneumoniae ApxI, ApxII, and ApxIII-specific ELISA methods for evaluation of vaccine efficacy / J Vet Sci 20:e2
Kishiro Y / 1995 / A novel method of preparing rat-monoclonal antibody-producing hybridomas by using rat medial iliac lymph node cells / Cell Struct Funct 20:151~156
Huang H / 2006 / Generation of monoclonal antibodies and epitope mapping of ApxIVA of Actinobacillus pleuropneumoniae / Mol Immunol 43:2130~2134
Inzana TJ / 2011 / Serologic detection of Actinobacillus pleuropneumoniae in swine by capsular polysaccharide-biotin-streptavidin enzyme-linked immunosorbent assay / J Clin Microbiol 39:1279~1282
Jung M / 2019 / Development of Actinobacillus pleuropneumoniae ApxI, ApxII, and ApxIII-specific ELISA methods for evaluation of vaccine efficacy / J Vet Sci 20:e2
Kishiro Y / 1995 / A novel method of preparing rat-monoclonal antibody-producing hybridomas by using rat medial iliac lymph node cells / Cell Struct Funct 20:151~156
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