Clinical Chemistry
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Clinical Chemistry 42: 1155-1158, 1996;
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Clinical Chemistry, Vol 42, 1155-1158, Copyright © 1996 by American Association for Clinical Chemistry

Direct determination of plasma endothelin-I by chemiluminescence enzyme immunoassay

R Iwata, T Hayashi, Y Nakao, M Yamaki, T Yoshimasa, H Ito, Y Saito, M Mukoyam and K Nakao
Pharmaceutical Research Laboratory, Hitachi Chemical Co., Ibaraki, Japan. HCN00974@niftyserve.or.jp

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.




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Copyright © 1996 by the American Association for Clinical Chemistry.