Cross-species reactivity of a monoclonal antibody against glutathione S-transferase fusion protein of human beta 2-adrenergic receptor

Biochem Mol Biol Int. 1998 Jun;45(2):215-25. doi: 10.1080/15216549800202582.

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-Ag14 cells and the resulting monoclonal antibody was named as mAb beta C02. The monoclonal antibody beta C02 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 C02 recognized human beta 2-adrenergic receptor in the beta 2-adrenergic receptor-GST fusion protein and human epidermoid carcinoma cell line A431 with highly specific immunoreactivity. In addition, mAb beta C02 showed cross-species reactivity against beta-adrenergic receptor of hamster lung and rat brain as revealed by Western blot and immunohistochemistry. The monoclonal antibody beta C02 may provide useful tools for the study of the beta-adrenergic receptor of human and other species including rats.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / isolation & purification
  • Antibody Specificity
  • Blotting, Western
  • Cricetinae
  • Enzyme-Linked Immunosorbent Assay
  • Glutathione Transferase / immunology
  • Humans
  • Immunoglobulin M / immunology
  • Immunoglobulin M / isolation & purification
  • Immunohistochemistry
  • Male
  • Mice
  • Mice, Inbred BALB C
  • Rats
  • Receptors, Adrenergic, beta-2 / immunology*
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / immunology
  • Species Specificity
  • Tumor Cells, Cultured

Substances

  • Antibodies, Monoclonal
  • Immunoglobulin M
  • Receptors, Adrenergic, beta-2
  • Recombinant Fusion Proteins
  • Glutathione Transferase