title: Direct determination of plasma endothelin-I by chemiluminescence enzyme immunoassay.
abstract: A highly sensitive sandwich-type chemiluminescence enzyme immunoassay for plasma endothelin-I (ET-1), involving no extraction steps, has been developed. Two populations of polyclonal antibodies were used in the present study: One is specific to the C-terminus of the endothelin family of peptides; the other, a Fab' fragment against the N-terminal core region of ET-1, is coupled with horseradish peroxidase (HRP). Labeled HRP activity was measured by using an enhanced chemiluminescence reaction of luminol/hydrogen peroxide. The assay was sensitive enough to detect 0.5 ng/L (0.05 pg/well) of plasma ET-1 and had no significant cross-reactivities with other related peptides, including endothelin-3 and the endothelin precursor peptide, big ET-1. Reliability of the assay was confirmed by comparison with an extraction-based procedure and a commercial kit.
authors.affiliation: Pharmaceutical Research Laboratory, Hitachi Chemical Co., Ibaraki, Japan. HCN00974@niftyserve.or.jp
authors.forename: R
authors.initials: R
authors.lastname: Iwata
authors.forename: T
authors.initials: T
authors.lastname: Hayashi
authors.forename: Y
authors.initials: Y
authors.lastname: Nakao
authors.forename: M
authors.initials: M
authors.lastname: Yamaki
authors.forename: T
authors.initials: T
authors.lastname: Yoshimasa
authors.forename: H
authors.initials: H
authors.lastname: Ito
authors.forename: Y
authors.initials: Y
authors.lastname: Saito
authors.forename: M
authors.initials: M
authors.lastname: Mukoyam
authors.forename: K
authors.initials: K
authors.lastname: Nakao
mesh_terms: D000328:Adult; D016232:Endothelins / Q000097:blood*; D006801:Humans; D007124:Immunoenzyme Techniques / Q000706:statistics & numerical data; D008163:Luminescent Measurements; D008875:Middle Aged; D011786:Quality Control; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D016232:Endothelins; D011933:Reagent Kits, Diagnostic