A multiplexed screening method for agonists and antagonists of the estrogen receptor protein

Abstract

The estrogen receptor (ER) is regarded as a significant drug target because of its important physical and pathological function. In this article, we describe a novel screening method to obtain agonists and antagonists of ER. ER was immobilized onto an aldehyde-modified glass slide. The affinity of Cy3-labeled estradiol for ER protein microarrays was then determined. Two libraries, one containing 29 synthetic compounds and the other with 384 natural products that served as a model, were screened to find new ligands for ER. The IC(50) values obtained for tamoxifen and raloxifene were consistent with those found in the literature (4.85 × 10(-7) M versus 1.74~4.23 × 10(-7) M and 7.58 × 10(-8) M versus 0.89~5.84 × 10(-8) M, respectively). Finally, 65 active ligands (5 synthetic compounds and 60 natural products) of ER were identified. This novel method gave identical results to a conventional fluorescence polarization assay, thus verifying the accuracy of this simultaneous multireceptor screening method based on protein microarrays. The presented method is sensitive, accurate, and reliable, and shows great potential for use in high-throughput drug-screening research.

Chemiluminescence detection with water-soluble iridium(iii) complexes containing a sulfonate-functionalised ancillary ligand

“…These compounds have commonly been used to sensitise the weak chemiluminescence oxidation of various organic compounds with strong inorganic oxidants such as cerium(IV), bromate and permanganate. [41][42][43][44][45] However, in this case, no signicant emission from either luminophore was observed, indicating that the postulated energy transfer pathway (Fig. 9b) does not make a signicant contribution to the chemiluminescence reaction of metal complexes.…”

Design, synthesis, in vitro and in silico evaluation of indole-based tetrazole derivatives as putative anti-breast cancer agents

“…This assay is based on the principle of detecting a change in fluorescence resulting from the competitive displacement of the fluorochrome-tagged estrogen by the investigated molecule from the ER-α-Fluormone™ ES2 complex. 50 If the compound does not have affinity for ER-α, it will not displace the Fluormone™ ES2 ligand, resulting in high fluorescence. Conversely, if the compound has a strong binding affinity for ER-α, it will displace the Fluormone™ ES2 ligand from ER-α, resulting in low fluorescence.…”

Solid-Phase Biological Assays for Drug Discovery

“…The miniaturized format provides a method for increased sample throughput and multiplexed analyte detection, and requires smaller reaction volumes, which allows for increased sample concentrations and reaction kinetics versus plate-based assays (9). In addition, microarrays can be printed into 96 well plates to allow integration with conventional liquid handling systems and can be imaged by a range of techniques, including colorimetry (69), fluorescence (70,71), SPR (72,73), and mass spectrometry (MS) (74,75), providing the ability to design a wide range of assays to study protein-ligand interactions.…”