Iwata R, Hayashi T, Nakao Y, Yamaki M, Yoshimasa T, Ito H, Saito Y, Mukoyam M, Nakao K
Pharmaceutical Research Laboratory, Hitachi Chemical Co., Ibaraki, Japan.
Clin Chem. 1996 Aug;42(8 Pt 1):1155-8.
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.
已开发出一种用于血浆内皮素-1（ET-1）的高灵敏度夹心型化学发光酶免疫测定法，该方法无需提取步骤。本研究使用了两种多克隆抗体：一种对内皮素肽家族的C末端具有特异性；另一种是针对ET-1 N末端核心区域的Fab'片段，与辣根过氧化物酶（HRP）偶联。通过使用鲁米诺/过氧化氢的增强化学发光反应来测量标记的HRP活性。该测定法灵敏度足以检测0.5 ng/L（0.05 pg/孔）的血浆ET-1，并且与其他相关肽，包括内皮素-3和内皮素前体肽大ET-1，没有明显的交叉反应。通过与基于提取的方法和商业试剂盒进行比较，证实了该测定法的可靠性。
