Abstract
This chapter describes procedures for the measurement of endothelin peptides by antibodies, focusing on the two main immunoassay techniques that are widely used. In a two-site “sandwich” enzyme-linked immunosorbent assay (ELISA) one antibody is immobilized to a solid phase and captures the endothelin (ET) peptide(s), which is quantified by the binding to this complex of a second, enzyme-labeled antibody in the liquid phase. In a radioimmunoassay (RIA), the ET peptide(s) to be measured competes for the binding of a fixed concentration of radiolabeled peptide to a fixed concentration of antibody in the liquid phase. In contrast to the ELISA, the immune complex measured in a RIA does not contain the analyte and therefore inverse (falling) standard curves of peptide concentration vs bound labeled peptide are produced. For a more detailed discussion of the two techniques, refs. 1 and 2 are recommended.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Gosling J.P. (ed.) (2000) ELISA Immunoassays. OUP, Oxford, UK.
Chard T. (1990) An Introduction to Radioimmunoassay and Related Techniques. (Burton R.H. and van Knippenberg P.H., eds.) Elsevier, Amsterdam, The Netherlands.
Davenport A.P. and Russell F.D. (2001) Endothelin converting enzymes and endothelin receptor localization in human tissues, in Handbook Exp. Pharmacol. vol.152 (Warner T.D., ed.), Springer-Verlag, Berlin, Germany, pp.209-237.
Russell F.D., Skepper J.N., and Davenport A.P. (1998) Human endothelial cell storage granules:a novel intracellular site for isoforms of the endothelin converting enzyme. Circ. Res. 83, 314–321.
Haynes W.G. and Webb D.J. (1994) Contribution of endogenous generation of endothelin-1 to basal vascular tone. Lancet 344, 852–854.
Bacon C.R., Cary N.R.B., and Davenport A.P. (1996) Endothelin peptide and receptors in human atherosclerotic coronary artery and aorta. Circ. Res. 79, 794–801.
Kurihara H., Yoshizumi M., Sugiyama T., Yamaoki K., Nagai R., Takaku F., Satih H., Inui J., Yanagisawa M., and Masaki T. (1989) The possible role of endothelin-1 in the pathogenesis of coronary vasospasm. J. Cardiovasc. Pharmacol. 13(Suppl 5), S132–S137.
Wei C.M., Lerman A., Rodeheffer R.J., McGregor C.G., Brandt R.R., Wright S., et al. (1994) Endothelin in human congestive heart failure. Circ. 89(4), 1580–1586.
Monge J.C. (1998) Neurohormonal markers of the clinical outcome of cardiovascular disease:is endothelin the best one? J. Cardiovasc. Pharmacol. 32(Suppl 2), S36–S42.
O’ Reilly G., Charnock-Jones D.S., Morrison J.J., Cameron I.T., Davenport A.P., and Smith S.K. (1993) Alternatively spliced mRNA’ s for human endothelin-2 and their tissue distribution. Biochem. Biophys. Res. Commum. 193, 834–840.
Plumpton C., Ashby M.J., Kuc R.E., O’ Reilly G., and Davenport A.P. (1996) Expression of endothelin peptides and mRNA in the human heart. Clin. Sci. 90, 37–46.
Maguire J.J. and Davenport A.P. (1995) ETA receptors mediate the constrictor responses to endothelin peptides in human blood vessels in vitro. Br. J. Pharmacol. 115, 191–197.
Howard P.G., Plumpton C., and Davenport A.P. (1992) Anatomical localization and pharmacological activity of mature endothelins and their precursors in human vascular tissue. J. Hypertens. 10, 1379–1386.
Matsumoto H., Suzuki N., Kitada C., and Fujino M. (1994) Endothelin family peptides in human plasma and urine:their molecular forms and concentrations. Peptides 15, 505–510.
Plumpton C., Champeney R., Ashby M.J., Kuc R.E., and Davenport A.P. (1993) Characterisation of endothelin isoforms in human heart:Endothelin-2 demonstrated. J. Cardiovasc. Pharmacol. 22(Suppl 8), 26–28.
Suzuki N., Matsumoto H., Miyauchi T., Goto K., Masaki T., Tsuda M., and Fujino M. (1990) Endothelin-3 concentrations in human plasma:increased concentrations in patients undergoing haemodialysis. Biochem. Biophys. Res. Commum. 169, 809–815.
Davenport A.P., Plumpton C., Ferro C.J., Webb D.J., and Horton J. (1998) Systemic infusion of an endothelin receptor antagonist increases plasma ET-3 in humans. Br. J. Pharmacol. 123, 290P.
Takahashi K., Ghatei M.A., Jones P.M., Murphey J.K., Lam H.C., O’Haloran D.J., and Bloom S.R. (1991) Endothelin in human brain and pituitary gland:presence of immunoreactive endothelin, endothelin messenger ribonucleic acid, and endothelin receptors. J. Clin. Endocrinol. Metab. 72, 693–699.
Plumpton C., Horton J., Kalinka K.S., Martin R. and Davenport A.P. (1994) Effects of phosphoramidon and pepstatin A on the secretion of endothelin-1 and big endothelin-1 in human umbilical vein endothelial cells:Measurement by two-site ELISAs. Clin. Sci. 87, 245–251.
Davenport A.P., Ashby M.J., Easton P., Ella S., Bedford J., Dickerson C., et al. (1990) A sensitive radioimmunoassay measuring endothelin-like immunoreactivity in human plasma:comparison of levels in essential hypertension and normotensive controls. Clin. Sci. 78, 261–264.
Plumpton C., Haynes W.G., Webb D.J., and Davenport A.P. (1995) Phosphoramidon inhibition of the in vivo conversion of big endothelin-1 to endothelin-1 in the human forearm. Br. J. Pharmacol. 115, 1821–1828.
Plumpton C., Haynes W.G., Webb D.J., and Davenport A.P. (1995) Measurement of C-terminal fragment of big endothelin-1:A novel method for assessing the generation of endothelin-1 in human. J. Cardiovasc. Pharmacol. 26(Suppl 3), S34–S36.
Plumpton C., Ferro C.J., Haynes W.G., Webb D.J., and Davenport A.P. (1996) The increase in human plasma immunoreactive endothelin but not big endothelin-1 or its C-terminal induced by systemic administration of the endothelin antagonist TAK-044. Br. J. Pharmacol. 119, 311–314.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2002 Humana Press Inc.
About this protocol
Cite this protocol
Davenport, A.P., Kuc, R.E. (2002). Analysis of Endothelins by Enzyme-Linked Immunosorbent Assay and Radioimmunoassay. In: Maguire, J.J., Davenport, A.P. (eds) Peptide Research Protocols. Methods in Molecular Biology™, vol 206. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-289-9:021
Download citation
DOI: https://doi.org/10.1385/1-59259-289-9:021
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-993-3
Online ISBN: 978-1-59259-289-0
eBook Packages: Springer Protocols