Preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of Lawsonia intracellularis
Porcine proliferative enteropathy (PPE), an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis, is commonly associated with diarrhea and reduced weight gain in growing pigs widespread. An accurate method for detecting L. intracellula...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | Journal of Integrative Agriculture |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S209531192300076X |
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| author | Ning XIAO Yun-yun LÜ Jian-nan LI Chang-feng CHEN Hui-xing LIN Hong-jie FAN |
| author_facet | Ning XIAO Yun-yun LÜ Jian-nan LI Chang-feng CHEN Hui-xing LIN Hong-jie FAN |
| author_sort | Ning XIAO |
| collection | DOAJ |
| description | Porcine proliferative enteropathy (PPE), an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis, is commonly associated with diarrhea and reduced weight gain in growing pigs widespread. An accurate method for detecting L. intracellularis is particularly important for preventing and controlling PPE. Heat shock protein 60 (Hsp60) is an immunodominant bacterial antigen found in all eukaryotic and prokaryotic organisms. Thus, the purpose of the current investigation was to produce a novel L. intracellularis Hsp60 monoclonal antibody (mAb) useful for immunodiagnostics. Three hybridomas secreted anti-Hsp60 termed 3E5, 4E2, and 9G6 were generated, and the titers of ascitic fluids of 3E5, 4E2, 9G6 were 1:1 024 000, 1:2 048 000 and 1:2 048 000, respectively. The Western blotting analysis demonstrated that recombinant Hsp60 (rHsp60) was recognized by mAbs 3E5, 4E2 and 9G6. Subsequently, analyses of specificity showed all the mAbs were highly specific to L. intracellularis while could not significantly react with other enteric bacteria commonly found in the ileum of pigs, such as Escherichia coli, Salmonella Choleraesuis, Salmonella Typhimurium, and Brachyspira hyodysenteriae. Furthermore, the mAbs were useful for detecting L. intracellularis in the infected monolayer cells and histological sections of the ileum from PPE-affected pigs. Our research will provide a foundation for the development of immunological diagnostic tests. |
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| format | Article |
| id | doaj.art-bc78c1219bf74753a8b34605a50da021 |
| institution | Directory Open Access Journal |
| issn | 2095-3119 |
| language | English |
| last_indexed | 2024-03-12T00:09:28Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Integrative Agriculture |
| spelling | doaj.art-bc78c1219bf74753a8b34605a50da0212023-09-16T05:30:32ZengElsevierJournal of Integrative Agriculture2095-31192023-09-0122928242833Preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of Lawsonia intracellularisNing XIAO0Yun-yun LÜ1Jian-nan LI2Chang-feng CHEN3Hui-xing LIN4Hong-jie FAN5MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R.ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R.ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R.ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R.ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R.ChinaMOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, P.R.China; Jiangsu Co-innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, P.R.China; Correspondence FAN Hong-jie, Tel/Fax: +86-25-84399592Porcine proliferative enteropathy (PPE), an important infectious disease in pig production caused by an obligate intracellular bacterium Lawsonia intracellularis, is commonly associated with diarrhea and reduced weight gain in growing pigs widespread. An accurate method for detecting L. intracellularis is particularly important for preventing and controlling PPE. Heat shock protein 60 (Hsp60) is an immunodominant bacterial antigen found in all eukaryotic and prokaryotic organisms. Thus, the purpose of the current investigation was to produce a novel L. intracellularis Hsp60 monoclonal antibody (mAb) useful for immunodiagnostics. Three hybridomas secreted anti-Hsp60 termed 3E5, 4E2, and 9G6 were generated, and the titers of ascitic fluids of 3E5, 4E2, 9G6 were 1:1 024 000, 1:2 048 000 and 1:2 048 000, respectively. The Western blotting analysis demonstrated that recombinant Hsp60 (rHsp60) was recognized by mAbs 3E5, 4E2 and 9G6. Subsequently, analyses of specificity showed all the mAbs were highly specific to L. intracellularis while could not significantly react with other enteric bacteria commonly found in the ileum of pigs, such as Escherichia coli, Salmonella Choleraesuis, Salmonella Typhimurium, and Brachyspira hyodysenteriae. Furthermore, the mAbs were useful for detecting L. intracellularis in the infected monolayer cells and histological sections of the ileum from PPE-affected pigs. Our research will provide a foundation for the development of immunological diagnostic tests.http://www.sciencedirect.com/science/article/pii/S209531192300076Xporcine proliferative enteropathyLawsonia intracellularismonoclonal antibodyimmunodiagnosticsheat shock protein 60 |
| spellingShingle | Ning XIAO Yun-yun LÜ Jian-nan LI Chang-feng CHEN Hui-xing LIN Hong-jie FAN Preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of Lawsonia intracellularis Journal of Integrative Agriculture porcine proliferative enteropathy Lawsonia intracellularis monoclonal antibody immunodiagnostics heat shock protein 60 |
| title | Preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of Lawsonia intracellularis |
| title_full | Preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of Lawsonia intracellularis |
| title_fullStr | Preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of Lawsonia intracellularis |
| title_full_unstemmed | Preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of Lawsonia intracellularis |
| title_short | Preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of Lawsonia intracellularis |
| title_sort | preparation and application of a novel monoclonal antibody specific for the heat shock protein 60 of lawsonia intracellularis |
| topic | porcine proliferative enteropathy Lawsonia intracellularis monoclonal antibody immunodiagnostics heat shock protein 60 |
| url | http://www.sciencedirect.com/science/article/pii/S209531192300076X |
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