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Clinical Chemistry 47: 102-109, 2001;
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(Clinical Chemistry. 2001;47:102-109.)
© 2001 American Association for Clinical Chemistry, Inc.

Articles

Use of Free Radical Chemistry in an Immunometric Assay for 17ß-Estradiol

Laure Buscarlet1, Hervé Volland3, Jacqueline Dupret-Carruel4, Michel Jolivet4, Jacques Grassi1, Christophe Créminon1, Frédéric Taran2 and Philippe Pradelles1,a

CEA,
1 Laboratoire d’Etudes Radioimmunologiques, SPI/DRM/DSV, and
2 Service des Molécules Marquées, DBCM/DSV, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France.
3 SPI-BIO, 2 rue du Buisson aux Fraises, ZI de la Bonde, 91741 Massy Cedex, France.

4 bioMérieux, Département Immunoassays, Chemin de l’Orme, 69280 Marcy-l’Etoile, France.
a Address correspondence to this author at: CEA, Laboratoire d’Etudes Radioimmunologiques, SPI/DRM/DSV, CEA/Saclay, 91191 Gif-sur-Yvette Cedex, France. Fax 33-01-69-86-77-49; philippe.pradelles{at}cea.fr

Background: We wished to develop an enzyme immunometric assay for 17ß-estradiol (E2) in human serum using solid-phase immobilized epitope immunoassay (SPIE-IA) technology and free radical chemistry.

Methods: We used an anti-estradiol monoclonal antibody as capture antibody and Fenton-like reagents to cross-link it to E2. The same antibody, labeled with acetylcholinesterase, was used for detection. Serum was diluted 10-fold before assay.

Results: After correction by the dilution factor, the detection limit was 5 ng/L for human serum and intra- and interassay CVs were <7% and 15%, respectively, at concentrations of 169-2845 ng/L. No cross-reactivity was seen with other natural steroids. In comparison with a competitive commercial RIA performed on 88 undiluted human sera, the slope (SD) of the regression line was 1.05 (± 0.02) and the intercept was 47 (±27) ng/L (Sy|x = 186 ng/L) at concentrations of 20–5000 ng/L (r2 = 0.97).

Conclusions: The use of Fenton-like chemistry in SPIE-IA technology allows a sensitive measurement of E2 in human serum and could be a new approach for the development of sensitive immunoassays.




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


Home page
 J. Clin. Endocrinol. Metab.Home page
F. Paris, N. Servant, B. Terouanne, P. Balaguer, J. C. Nicolas, and C. Sultan
A New Recombinant Cell Bioassay for Ultrasensitive Determination of Serum Estrogenic Bioactivity in Children
J. Clin. Endocrinol. Metab., February 1, 2002; 87(2): 791 - 797.
[Abstract] [Full Text] [PDF]


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