An advanced biosensor for the prediction of estrogenic effects of endocrine-disrupting chemicals on the estrogen receptor alpha.
A label-free and time-resolved biosensor based on reflectometric interference spectroscopy (RIfS) has been developed to evaluate the agonistic or antagonistic effects of potential ligands with unknown behavior. The biosensor utilizes the specific interaction between the estrogen receptor alpha (ER alpha) and short specific peptides. The unique feature of these peptides allows the investigation of the behavior of ligands and the discrimination between the agonistic and antagonistic effects caused by conformational changes of the receptor. Thus, this developed biosensor allows not only the differentiation between ligands and nonligands of a receptor, but also the potential of these ligands to influence conformational changes in the receptor, leading to activation or inhibition of the receptor-dependent pathways. Owing to the robustness of the direct optical detection principle used, the biosensor is applicable to complex biological matrices, even crude cell extracts. Moreover, the reliability of the biosensor, including regeneration steps when performing subsequent measurements, has been verified.
Fechner
Peter
P
Institute of Physical and Theoretical Chemistry, Eberhard Karls University of Tübingen, Auf der Morgenstelle 8, 72076, Tübingen, Germany. peter.fechner@ipc.uni-tuebingen.de
Pröll
Florian
F
Carlquist
Mats
M
Proll
Günther
G
D001665:Binding Sites; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D052244:Endocrine Disruptors / Q000494:pharmacology*; D004958:Estradiol / Q000737:chemistry; D047628:Estrogen Receptor alpha / Q000187:drug effects*; D006801:Humans; D007368:Interferometry / Q000295:instrumentation / Q000379:methods; D008024:Ligands; D008958:Models, Molecular; D010047:Ovalbumin / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D011237:Predictive Value of Tests; D011487:Protein Conformation; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013057:Spectrum Analysis / Q000295:instrumentation / Q000379:methods; D013499:Surface Properties; D013629:Tamoxifen / Q000737:chemistry; D013997:Time Factors
D052244:Endocrine Disruptors; D047628:Estrogen Receptor alpha; D008024:Ligands; D010455:Peptides; D013629:Tamoxifen; D011092:Polyethylene Glycols; D004958:Estradiol; D010047:Ovalbumin; D004121:Dimethyl Sulfoxide