Characterization and application of monoclonal antibodies against Mycoplasma hyorhinis pyruvate dehydrogenase E1 complex subunit alpha

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• 作者：Dongjie Chen ; Yanwu Wei ; Liping Huang…
• 关键词：Mycoplasma hyorhinis ; Pyruvate dehydrogenase E1 complex subunit alpha ; Monoclonal antibody ; B cell epitopes ; Immunohistochemistry
• 刊名：Applied Microbiology and Biotechnology
• 出版年：2016
• 出版时间：April 2016
• 年：2016
• 卷：100
• 期：8
• 页码：3587-3597
• 全文大小：1,434 KB
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• 作者单位：Dongjie Chen (1)
  Yanwu Wei (1)
  Liping Huang (1)
  Yiping Wang (1)
  Wenjuan Du (1)
  Jianhui Sun (1)
  Hongli Wu (1)
  Li Feng (1)
  Changming Liu (1)
  
  1. Division of Swine Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, 427 Maduan Street, Nangang District, Harbin, 150001, People’s Republic of China
  
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chemistry
  Biotechnology
  Microbiology
  Microbial Genetics and Genomics
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-0614

文摘

Mycoplasma hyorhinis is commonly found in the respiratory tract of pigs and is the etiological agent of polyserositis. The metabolic enzymes of M. hyorhinis may play important roles in host-pathogen interactions. We immunized BALB/c mice with sodium deoxycholate-extracted antigens (DOC-Ags) and screened 10 hybridomas that secreted antibodies against various M. hyorhinis proteins. Pyruvate dehydrogenase E1 complex subunit alpha (PDHA) was identified as the protein that reacted with five of the 10 monoclonal antibodies (mAbs). Sequence analysis indicated that PDHA was highly conserved among M. hyorhinis strains, but not among other mycoplasmas. We predicted the three-dimensional structure of PDHA and identified three epitopes (277RTEEEEK283, 299KDKKYITDE307, and 350LKEQKQHAKDY360). The mAb 1H12 we generated was used to detect M. hyorhinis PDHA in vitro and in piglets infected with M. hyorhinis. We observed that PDHA was mainly located in the epithelial cells of the lungs. Our results indicate that the mAbs we generated could be used to further investigate the structure and function of M. hyorhinis PDHA. In addition, they could be used in the differential diagnosis of M. hyorhinis and other mycoplasmas.