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Biosensors for Detecting Estrogen-like Molecules and Protein Biomarkers

  • James L. Wittliff
  • Sarah A. Andres
  • Traci L. Kruer
  • D. Alan Kerr
  • Irina A. Smolenkova
  • Judith L. Erb
Part of the Advances In Experimental Medicine And Biology book series (AEMB, volume 614)

Abstract

A novel evanescent-based biosensor (Endotect™, ThreeFold Sensors, Inc.) was developed with laser-based fiber optics using fluorescent dyelabeled recombinant human estrogen receptor-α (rhERα) and hERβ as probes. A three-tiered approach evaluating various steps in the formation of the estrogen- receptor complex and its subsequent activity was developed for instrument calibration to detect estrogen mimics in biological samples, water and soil. Using this approach, binding affinities and activities of certain known estrogen mimics were determined for their use as calibrator molecules. Results indicated rhERα and rhERβ may be employed as probes to distinguish estrogen mimics with a broad range of affinities. In addition, application of the biosensor for detecting DNA-binding proteins in human tissue extracts was demonstrated. The later studies suggest the biosensor may be used as a clinical laboratory tool for assessing tumor marker proteins.

Keywords

Electrophoretic Mobility Shift Assay Estrogen Response Element Evanescent Field Human Estrogen Receptor Estrogen Receptor Protein 
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. 1.
    ICCVAM/NICEATM Final Report, Expert Panel Evaluation of the Validation Status of in vitro Test Methods for Detecting Endocrine Disruptors: Estrogen Receptor and Androgen Receptor Binding and Transcriptional Activation Assays, 2002 (http:// iccvam.niehs.nih.gov).http://iccvam.niehs.nih.gov).Google Scholar
  2. 2.
    J. L. Wittliff, R. Pasic, K. I. Bland: Steroid and Peptide Hormone Receptors: Methods, Quality Control and Clinical Use, in Bland KI, Copeland III EM (eds): The Breast: Comprehensive Management of Benign and Malignant Diseases. Philadelphia, PA, W. B. Saunders Co, 458–498, 1998.Google Scholar
  3. 3.
    R. H. Smith, W. J. Lemon, J. L. Erb, J. R. Erb-Downward, J. G. Downward, O. E. Ulrich, and J. L. Wittliff, Development of Kinetic Ligand-binding Assays Using a Fiber Optic Sensor, Clin. Chem. 45(9), 1683–1685 (1999).PubMedGoogle Scholar
  4. 4.
    E. A. E. Garber, J. L. Erb, J. G. Downward, E. M. Priuska, J. L. Wittliff, W. Feng, J. Magner, and G. L. Larsen, Biosensor, ELISA, and Frog Embryo Teratogenesis Assay: Xenopus (FETAX) Analysis of Water Associated with Frog Malformations in Minnesota, Proc. Soc. Photo-Optical Instrumentation Engineers (SPIE) 4206, 147–158 (2001).Google Scholar
  5. 5.
    J. L. Erb, E. A. E. Garber, J. G. Downward IV, and E. M. Priuska, Data from an Estrogen Receptor-based Biosensor Correlates with Evidence of Frog Malformation and Demonstrates a Differential Response of hERα & β to Beneficial and Harmful Estrogenic Compounds, In: Proc. 2nd Intl. Conf. Pharmaceuticas & Endocrine Disrupting Chemicals in Water, p. 203–217, Westerville, OH, The National Ground Water Association (2001).Google Scholar
  6. 6.
    T. E. Hirchfield, Fluorescent Immunoassay Employing Optical Fiber in a Capillary Tube, U.S. Patent No. 4,447,546 (1984).Google Scholar
  7. 7.
    W. Raffelsberger and J. L. Wittliff, A Novel Approach for Comparing Ligand Binding Results from Titration and Competition Analyses to Study Hormone Mimics, J. Clin. Ligand Assay 20(4), 275–280 (1997).Google Scholar
  8. 8.
    J. L. Wittliff, L. L. Wenz, J. Dong, Z. Nawaz, and T. R. Butt, Expression and Characterization of an Active Estrogen Receptor as a Ubiquitin Fusion Protein from Escherichia coli, J. Biol. Chem. 265(35), 22016–22025 (1990).PubMedGoogle Scholar
  9. 9.
    K. Graumann, J. L. Wittliff, W. Raffelsberger, L. Miles, A. Jungbauer, and T.R. Butt, Structural and Functional Analysis of N-terminal Point Mutants of the Human Estrogen Receptor, J. Steroid Biochem. Mol. Biol. 57(5–6), 292–300 (1996).Google Scholar
  10. 10.
    C. R. Lyttle, P. Damian-Matsumura, H. Juul, and T. R. Butt, Human Estrogen Receptor Regulation in a Yeast Model System and Studies on Receptor Agonists and Antagonists, J. Steroid Biochem. Mol. Biol. 42(7), 77–685 (1992).CrossRefGoogle Scholar
  11. 11.
    J. L. Wittliff, P. Folk, J. Dong, C. Schaupp, and T. R. Butt, Characteristics of the Human Estrogen Receptor Protein Produced in Microbial Expression Systems. In: V. K. Moudgil (ed.), Steroid Hormone Receptors: Basic and Clinical Aspects, pp. 473–501, Birkhauser Boston (1993).Google Scholar
  12. 12.
    T. L. Kruer, I. A. Smolenkova and J. L. Wittliff, Expression of Novel ERE-Binding Proteins in Breast and Uterine Cells, American Society for Biochemistry & Molecular Biology, Abstract # 1381 (2006).Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • James L. Wittliff
    • 1
  • Sarah A. Andres
    • 1
  • Traci L. Kruer
    • 1
  • D. Alan Kerr
    • 1
  • Irina A. Smolenkova
    • 1
  • Judith L. Erb
    • 2
  1. 1.Department of Biochemistry & Molecular BiologyInstitute for Molecular Diversity & Drug Design, University of LouisvilleLouisvilleUSA
  2. 2.IA, Inc./ThreeFold SensorsAnn ArborUSA

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