Production and Characterization of a Monoclonal Antibody against Human ${\beta}_2$-adrenergic receptor

  • Kang, Suk-Jo (College of Pharmacy and Institute for Molecular Bilolgy and Genetics, Seoul National Uneversity) ;
  • Shin, Chan-Young (College of Pharmacy and Institute for Molecular Bilolgy and Genetics, Seoul National Uneversity) ;
  • Song, Mi-Ryoung (College of Pharmacy and Institute for Molecular Bilolgy and Genetics, Seoul National Uneversity) ;
  • Lee, Chung-Jae (College of Pharmacy and Institute for Molecular Bilolgy and Genetics, Seoul National Uneversity) ;
  • Cheong, Jae-Hoon (Department of Pharmacy, Samyook University) ;
  • Lee, Sang-Bong (Signal Transduction Research Unit, Korea Tesearch Institute of Bioscience and Biotechnology) ;
  • Ko, Kwang-Ho (College of Pharmacy and Institute for Molecular Bilolgy and Genetics, Seoul National Uneversity)
  • Published : 1997.12.01

Abstract

The purpose of the present study was to produce and characterize a monoclonal antibody against human ${\beta}_2$-adrenergic receptor. Male BALB/c mice were immunized with glutathione S-transferase (GST) fusion protein of the C-terminal portion of the human ${\beta}_2$-adrenergic receptor which was expressed in E.Coli. The immunized splenocytes were fused with myeloma SP2/0-Agl4 cells. The resulting hybridomas were screened for the production of a monoclonal antibody which can recognize human ${\beta}_2$-adrenergic receptor, and then subcloned by limiting dilution. The resulting monoclonal antibody was named as mAb$\beta$CO2. The mono-clonal antibody $\beta$CO2 was determined as IgM subtype and then purified by anti-mouse IgM-agarose affinity chromatography. The results of ELISA, Western blot, and immunocytochemistry showed that mAb$\beta$CO2 recognized human ${\beta}_2$-adrenergic receptor in the ${\beta}_2$-adrenergic receptor-GST fusion protein and human spider-moid carcinoma cell line A431 with highly specific immunoreactivity, The monoclonal antibody $\beta$CO2 may provide useful tools for the study of the $\beta$-adrenergic receptor of human and other species including rats.

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