Radio-immunoassay of endothelin in human plasma

Abstract

Endothelin (ET) is a newly described endothelium-derived 21-amino-acid peptide with potent vasoconstrictive properties. The present study describes a radio-immunoassay utilizing an antibody without cross-reactivity with big endothelin, produced after immunization of rabbits with endothelin-1 (ET-1). Measurement of endothelin in human plasma was performed after prior extraction on Sep-pak C₁₈ cartridges. Recovery of unlabelled endothelin-1 added to human plasma (3.5 pg ml^-1) was 84.8 ± 11.1% (mean ± SD, n=12). The minimum detectable level in plasma was 50 fmol l^-1. High-pressure liquid chromatography on reverse-phase C₁₈ column established that the measured irET eluted identically to ET-1. The concentration of irET in plasma from healthy control subjects was 1.11 ± 0.2 pmol^-1 (mean ± SD, n=30). Agarose electrophoresis of plasma with [¹²⁵I]-ET-1 indicated a reversible binding to albumin. It is concluded that ET-1 is present in measurable amounts in human plasma. It is suggested that ET-1 in plasma is associated with albumin. The importance of an adequate extraction procedure is stressed.

Key Words:

• endothelin
• HPLC
• radio-immunoassay
• reverse-phase chromatography
• vasoconstriction