In Vitro Analysis of Xenoestrogens by Enzyme Linked Receptor Assays (ELRA)

  • Martin Seifert
  • Stefanie Haindl
  • Bertold Hock
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 444)

Abstract

Receptor binding assays are a modern approach to the effects-related analysis of xenoendocrines. Human estrogen receptors are used to develop an analytical tool for the detection of estrogenic substances in environmental samples. A sensitive Enzyme Linked Receptor Assay (ELRA) was developed in a microwell plate format. The receptor assay is based on similar principles as competitive immunoassays (ELISA). However, receptor binding always implies a biological effect, either agonistic or antagonistic. The choice of suitable tracers or hapten conjugates is an important step in assay development. For this purpose and for the determination of receptor-affinities of relevant xenoestrogens, a surface plasmon resonance (SPR) biosensor (BIAcore) was used for binding studies with immobilized receptors. Results with commercially available hapten-conjugates (estradiol-BSA) show a direct correlation between the amount of immobilized estradiol receptor and the amount of bound hapten-conjugate. Based on these BIAcore experiments an ELRA was developed. The calibration curves show a detection limit of 0.1 μg/1 for 17β-estradiol. Cross-reactivities of different steroids and xenoestrogens are reported.

Keywords

Surface Plasmon Resonance Testicular Cancer Synthetic Estrogen Receptor Assay Surface Plasmon Resonance Biosensor 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Sumpter, J. P., Joblin, S., 1995, Environ. Health. Perspect. 103 (Suppl 7): 173–178PubMedGoogle Scholar
  2. Bull, J.J., Gutzke, W.H.N., Crews, D., 1988, Gen. Com. Endocrinol. 70: 425–428.CrossRefGoogle Scholar
  3. Carlsen, E., Giwercman, A., Keiding, N., Skakkebæk, N.E., 1995, Environ. Health. Perspect. 103 (Suppl. 7): 137–139.PubMedGoogle Scholar
  4. Godden, J., Leake, R., Kerr, D.J., 1992, Anticancer Research 12: 1683–1688.PubMedGoogle Scholar
  5. Hock, B., Seifert, M., 1997, Monitoring of effects, 202–223, in: Biosensors for Environmental Monitoring, Editors: B. Hock, D. Barceló, K. Cammann, P.D. Hansen, A.P.F. Turner, B.G. Teubner, Stuttgart (in press).Google Scholar
  6. Neumann, F., Schenk, B., 1991, Endokrinpharmakologie, 414–417; in: Pharmacologie und Toxicologie, Editors: W. Forth, D. Henschler, W. Rummel, BI-Wissenschaftsverlag.Google Scholar
  7. Long, K.P., 1997, Crit. Rev. Food. Sci. Nutr. 37(2): 93–209.CrossRefGoogle Scholar
  8. Kurzer, M.S., Xu, X., 1997, Annu. Rev. Nutr. 17: 353–381.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • Martin Seifert
    • 1
  • Stefanie Haindl
    • 1
  • Bertold Hock
    • 1
  1. 1.Department of BotanyTechnical University of MuenchenFreising/WeihenstephanGermany

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