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Clinical Chemistry 43: 64-70, 1997;
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(Clinical Chemistry. 1997;43:64-70.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Sandwich-type enzyme immunoassay for big endothelin-1 in plasma: concentrations in healthy human subjects unaffected by sex or posture

Philippe Aubin1, Gaelle Le Brun1, Florina Moldovan1, Jean-Marie Villette1, Christophe Créminon2, Jacques Dumas3, Laurence Homyrda1, Hany Soliman1, Michel Azizi4 and Jean Fiet1,a

1 Laboratoire de Biologie Hormonale, Hôpital Saint-Louis, 75010 Paris, France.

2 CEA Saclay, Service de Pharmacologie et d'Immunologie, 91191 Gif/Yvette, France.

3 Biotechnology Department, Roussel-Uclaf, 93235 Romainville, France.

4 Centre d'Investigation Clinique, INSERM, Hôpital Broussais, 75014 Paris, France.
a Author for correspondence. Fax +33 1 42494280.

A sandwich-type enzyme immunoassay has been developed for measuring human big endothelin-1 (big ET-1) in human plasma and supernatant fluids from human cell cultures. Big ET-1 is the precursor of endothelin 1 (ET-1), the most potent vasoconstrictor known. A rabbit antibody raised against the big ET-1 COOH-terminus fragment was used as an immobilized antibody (anti-P16). The Fab' fragment of a monoclonal antibody (1B3) raised against the ET-1 loop fragment was used as the enzyme-labeled antibody, after being coupled to acetylcholinesterase. The lowest detectable value in the assay was 1.2 pg/mL (0.12 pg/well). The assay was highly specific for big ET-1, demonstrating no cross-reactivity with ET-1, <0.4% cross-reactivity with big endothelin-2 (big ET-2), and <0.1% with big endothelin-3 (big ET-3). We used this assay to evaluate the effect of two different postural positions (supine and standing) on plasma big ET-1 concentrations in 11 male and 11 female healthy subjects. Data analysis revealed that neither sex nor body position influenced plasma big ET-1 concentrations. This assay should thus permit the detection of possible variations in plasma concentrations of big ET-1 in certain pathologies and, in association with ET-1 assay, make possible in vitro study of endothelin-converting enzyme activity in cell models. Such studies could clarify the physiological and clinical roles of this family of peptides.


Key Words: indexing terms: reference values • endothelins • isopeptides • vasoconstrictors • blood pressure • variation, source of




The following articles in journals at HighWire Press have cited this article:


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J. Papassotiriou, N. G. Morgenthaler, J. Struck, C. Alonso, and A. Bergmann
Immunoluminometric Assay for Measurement of the C-Terminal Endothelin-1 Precursor Fragment in Human Plasma
Clin. Chem., June 1, 2006; 52(6): 1144 - 1151.
[Abstract] [Full Text] [PDF]

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C. ODOUX, B. CRESTANI, G. LEBRUN, C. ROLLAND, P. AUBIN, N. SETA, M.-F. KAHN, J. FIET, and M. AUBIER
Endothelin-1 Secretion by Alveolar Macrophages in Systemic Sclerosis
Am. J. Respir. Crit. Care Med., November 1, 1997; 156(5): 1429 - 1435.
[Abstract] [Full Text]

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 Clin. Chem.Home page
V. Mathew and A. Lerman
Clinical Implications of a Sandwich Enzyme Immunoassay for Big Endothelin-1
Clin. Chem., January 1, 1997; 43(1): 9 - 10.
[Full Text] [PDF]

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 J. Biol. Chem.Home page
S. Lopez-Ongil, V. Senchak, M. Saura, C. Zaragoza, M. Ames, B. Ballermann, M. Rodriguez-Puyol, D. Rodriguez-Puyol, and C. J. Lowenstein
Superoxide Regulation of Endothelin-converting Enzyme
J. Biol. Chem., August 18, 2000; 275(34): 26423 - 26427.
[Abstract] [Full Text] [PDF]


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Copyright © 1997 by the American Association for Clinical Chemistry.