Cross-species reactivity of a monoclonal antibody against glutathione S-transferase fusion protein of human beta 2-adrenergic receptor.
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.
College of Pharmacy, Seoul National University, Korea.
C Y
CY
Shin
S J
SJ
Kang
M R
MR
Song
K H
KH
Park
D O
DO
Seo
J H
JH
Cheong
K H
KH
Ko
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D015153:Blotting, Western; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D005982:Glutathione Transferase / Q000276:immunology; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology / Q000302:isolation & purification; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D051381:Rats; D018343:Receptors, Adrenergic, beta-2 / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D013045:Species Specificity; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D007075:Immunoglobulin M; D018343:Receptors, Adrenergic, beta-2; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase