Endothelin in human plasma and culture medium of aortic endothelial cells--detection and characterization with radioimmunoassay using monoclonal antibody.
We have developed monoclonal (KY-ET-1-I) and polyclonal (ET-F5) antibodies against endothelin-1 (ET-1) and established sensitive radioimmunoassays (RIAs) with different specificities. The RIA with KY-ET-1-I detected ET-1, ET-2 and ET-3, while the RIA with ET-F5 recognized ET-3 very weakly. Using these RIAs, we have investigated the concentration and molecular forms of ET-1-like immunoreactivity (-LI) in culture medium of bovine aortic endothelial cells and human plasma. Culture medium of endothelial cells contained two major components compatible with big ET and ET-1. ET-1-LI was also detected in human plasma. ET-1-LI in human plasma consisted of apparent two components, the small molecular form emerging at the position of ET-1 and the large form with the peak eluting at the preceding fraction of the elution position of big ET. The concentration of the small form of ET in human plasma was about 5 pg/ml.
Saito
Y
Y
Department of Medicine, Kyoto University School of Medicine, Japan.
Nakao
K
K
Itoh
H
H
Yamada
T
T
Mukoyama
M
M
Arai
H
H
Hosoda
K
K
Shirakami
G
G
Suga
S
S
Jougasaki
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis; D001011:Aorta; D001202:Ascitic Fluid / Q000032:analysis; D002417:Cattle; D002850:Chromatography, Gel; D003470:Culture Media / Q000032:analysis*; D016232:Endothelins; D004730:Endothelium, Vascular / Q000032:analysis* / Q000166:cytology; D005260:Female; D007106:Immune Sera / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000097:blood* / Q000276:immunology / Q000302:isolation & purification; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D003470:Culture Media; D016232:Endothelins; D007106:Immune Sera; D010455:Peptides