Preparation and Biological Activity of the Monoclonal Antibody against the Second Extracellular Loop of the Angiotensin II Type 1 ReceptorReportar como inadecuado




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Journal of Immunology Research - Volume 2016 2016, Article ID 1858252, 10 pages -

Research Article

Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Capital Medical University, Beijing 100069, China

Beijing Key Laboratory of Metabolic Disorders Related Cardiovascular Diseases, Capital Medical University, Beijing 100069, China

Department of Physiology, Basic Medical Department, Fenyang College of Shanxi Medical University, Fenyang, Shanxi 032200, China

Department of Pathology, Shanxi Medical University, Taiyuan, Shanxi 030001, China

Department of Emergency Medicine, Thomas Jefferson University, 1025 Walnut Street, College Building, Suite 808, Philadelphia, PA 19107, USA

Received 12 October 2015; Accepted 27 December 2015

Academic Editor: Xiao-Feng Yang

Copyright © 2016 Mingming Wei et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

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 × 10

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.





Autor: Mingming Wei, Chengrui Zhao, Suli Zhang, Li Wang, Huirong Liu, and Xinliang Ma

Fuente: https://www.hindawi.com/



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