Biosensors for detecting estrogen-like molecules and protein biomarkers.
A novel evanescent-based biosensor (Endotect, ThreeFold Sensors, Inc.) was developed with laser-based fiber optics using fluorescent dye-labeled recombinant human estrogen receptor-alpha (rhERalpha) and hERbeta 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 rhERalpha and rhERbeta 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.
Wittliff
James L
JL
Department of Biochemistry & Molecular Biology, Institute for Molecular Diversity & Drug Design, University of Louisville, Louisville, KY, USA.
Andres
Sarah A
SA
Kruer
Traci L
TL
Kerr
D Alan
DA
Smolenkova
Irina A
IA
Erb
Judith L
JL
D015415:Biomarkers / Q000378:metabolism; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D002138:Calibration; D002232:Carbocyanines / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D047628:Estrogen Receptor alpha / Q000235:genetics / Q000378:metabolism; D047629:Estrogen Receptor beta / Q000235:genetics / Q000378:metabolism; D004967:Estrogens / Q000032:analysis* / Q000378:metabolism; D005336:Fiber Optic Technology / Q000295:instrumentation; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007834:Lasers; D018716:Molecular Mimicry; D015335:Molecular Probes; D055100:Optical Fibers; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D011869:Radioligand Assay; D011994:Recombinant Proteins / Q000378:metabolism
D015415:Biomarkers; D002232:Carbocyanines; D004268:DNA-Binding Proteins; D047628:Estrogen Receptor alpha; D047629:Estrogen Receptor beta; D004967:Estrogens; D005456:Fluorescent Dyes; D015335:Molecular Probes; D011506:Proteins; D011994:Recombinant Proteins; C085321:cyanine dye 5