Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 Receptor
The current study was to prepare a mouse-derived antibody against the angiotensin II type 1 receptor (AT1-mAb) based on monoclonal antibody technology, to provide a foundation for research on AT1-AA-positive diseases. Balb/C mice were actively immunized with the second extracellular loop of the angi...
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| Format: | Article |
| Language: | English |
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Wiley
2016-01-01
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| Series: | Journal of Immunology Research |
| Online Access: | http://dx.doi.org/10.1155/2016/1858252 |
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| author | Mingming Wei Chengrui Zhao Suli Zhang Li Wang Huirong Liu Xinliang Ma |
| author_facet | Mingming Wei Chengrui Zhao Suli Zhang Li Wang Huirong Liu Xinliang Ma |
| author_sort | Mingming Wei |
| collection | DOAJ |
| description | The current study was to prepare a mouse-derived antibody against the angiotensin II type 1 receptor (AT1-mAb) based on monoclonal antibody technology, to provide a foundation for research on AT1-AA-positive diseases. Balb/C mice were actively immunized with the second extracellular loop of the angiotensin II type 1 receptor (AT1R-ECII). Then, mouse spleen lymphocytes were fused with myeloma cells and monoclonal hybridomas that secreted AT1-mAb were generated and cultured, after which those in logarithmic-phase were injected into the abdominal cavity of mice to retrieve the ascites. Highly purified AT1-mAb was isolated from mouse ascites after injection with 1 × 107 hybridomas. A greater amount of AT1-mAb was purified from mouse ascites compared to the cell supernatant of hybridomas. AT1-mAb purified from mouse ascites constricted the thoracic aorta of mice and increased the beat frequency of neonatal rat myocardial cells via the AT1R, identical to the effects of AT1-AA extracted from patients’ sera. Murine blood pressure increased after intravenous injection of AT1-mAb via the tail vein. High purity and good biological activity of AT1-mAb can be obtained from mouse ascites after intraperitoneal injection of monoclonal hybridomas that secrete AT1-mAb. These data provide a simple tool for studying AT1-AA-positive diseases. |
| format | Article |
| id | doaj-art-6d679d756abb40b9b944e387a14d4dc3 |
| institution | OA Journals |
| issn | 2314-8861 2314-7156 |
| language | English |
| publishDate | 2016-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Journal of Immunology Research |
| spelling | doaj-art-6d679d756abb40b9b944e387a14d4dc32025-08-20T02:06:12ZengWileyJournal of Immunology Research2314-88612314-71562016-01-01201610.1155/2016/18582521858252Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 ReceptorMingming Wei0Chengrui Zhao1Suli Zhang2Li Wang3Huirong Liu4Xinliang Ma5Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, ChinaDepartment of Physiology, Basic Medical Department, Fenyang College of Shanxi Medical University, Fenyang, Shanxi 032200, ChinaDepartment of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, ChinaDepartment of Pathology, Shanxi Medical University, Taiyuan, Shanxi 030001, ChinaDepartment of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, ChinaDepartment of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, ChinaThe current study was to prepare a mouse-derived antibody against the angiotensin II type 1 receptor (AT1-mAb) based on monoclonal antibody technology, to provide a foundation for research on AT1-AA-positive diseases. Balb/C mice were actively immunized with the second extracellular loop of the angiotensin II type 1 receptor (AT1R-ECII). Then, mouse spleen lymphocytes were fused with myeloma cells and monoclonal hybridomas that secreted AT1-mAb were generated and cultured, after which those in logarithmic-phase were injected into the abdominal cavity of mice to retrieve the ascites. Highly purified AT1-mAb was isolated from mouse ascites after injection with 1 × 107 hybridomas. A greater amount of AT1-mAb was purified from mouse ascites compared to the cell supernatant of hybridomas. AT1-mAb purified from mouse ascites constricted the thoracic aorta of mice and increased the beat frequency of neonatal rat myocardial cells via the AT1R, identical to the effects of AT1-AA extracted from patients’ sera. Murine blood pressure increased after intravenous injection of AT1-mAb via the tail vein. High purity and good biological activity of AT1-mAb can be obtained from mouse ascites after intraperitoneal injection of monoclonal hybridomas that secrete AT1-mAb. These data provide a simple tool for studying AT1-AA-positive diseases.http://dx.doi.org/10.1155/2016/1858252 |
| spellingShingle | Mingming Wei Chengrui Zhao Suli Zhang Li Wang Huirong Liu Xinliang Ma Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 Receptor Journal of Immunology Research |
| title | Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 Receptor |
| title_full | Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 Receptor |
| title_fullStr | Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 Receptor |
| title_full_unstemmed | Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 Receptor |
| title_short | Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 Receptor |
| title_sort | preparation and biological activity of the monoclonal antibody against the second extracellular loop of the angiotensin ii type 1 receptor |
| url | http://dx.doi.org/10.1155/2016/1858252 |
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