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 angiotensin ii type 1 receptor (at-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 at,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. © 2016 mingming wei et al.