A sensitive sandwich-enzyme immunoassay for human endothelin.
A sensitive enzyme immunoassay (EIA) for human endothelin(1-21) has been established. The assay is based on a sandwich method that uses two differing capture and detection anti-endothelin antibodies. A monoclonal anti-endothelin antibody AwETN40, which did not react with an endothelin C-terminal heptapeptide, was used as an immobilized antibody. The Fab' fragment of rabbit antibodies against the endothelin C-terminal heptapeptide was used as an enzyme-labeled detector antibody after being coupled with horseradish peroxidase (HRP). The assay is sensitive enough to detect as little as 0.2 pg/well (80 amol/well) of endothelin. Preliminary investigations indicated that the basal level of immunoreactive endothelin in male plasma (n = 24) extracted with Seppak C-18 cartridges was 1.59 +/- 0.32 pg/ml.
Suzuki
N
N
Tsukuba Research Laboratories, Takeda Chemical Industries, Ltd., Ibaraki, Japan.
Matsumoto
H
H
Kitada
C
C
Masaki
T
T
Fujino
M
M
D000328:Adult; D000818:Animals; D000911:Antibodies, Monoclonal; D000937:Antigen-Antibody Reactions; D001667:Binding, Competitive; D016232:Endothelins; D004730:Endothelium, Vascular / Q000032:analysis*; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008875:Middle Aged; D009500:Neutralization Tests; D010455:Peptides / Q000032:analysis* / Q000097:blood / Q000276:immunology; D011817:Rabbits
D000911:Antibodies, Monoclonal; D016232:Endothelins; D010455:Peptides