A novel analytical tool for quantification of estrogenicity in river water based on fluorescence labelled estrogen receptor alpha

作者：Le Blanc Alexander Fabian^*; Albrecht Christiane; Bonn Tomas; Fechner Peter; Proll Guenther; Proell Florian; Carlquist Mats; Gauglitz Guenter

来源：Analytical and Bioanalytical Chemistry, 2009, 395(6): 1769-1776.

DOI：10.1007/s00216-009-3038-8

• Estrogen receptor alpha
• Advanced protein labelling
• Endocrine-disrupting chemicals (EDC)
• River water
• Environmental monitoring
• Response-related analytics

A novel combined procedure for estrogen-affinity purification and labelling of estrogen receptor alpha ligand-binding domain with Cy (TM) 5.5 cystein reactive dye was established. By using this procedure, mainly functional proteins are recovered. It can be easily adapted to a large variety of other proteins for which ligand-coated affinity materials are available. The labelled receptor was used in a total internal reflection fluorescence-based binding inhibition assay for determination of the impact of pollutants in river water on the receptor. The great advantage compared to conventional methods is that the total effect on the receptor is measured instead of concentrations of single compounds and that even currently unknown ligands are found as well. Therefore, the obtained signal is related to the response of the organism, which is exposed to the water. The limit of detection was found to be 0.139 nM of estradiol equivalents. The assay also provides a highly sensitive tool for pharmaceutical research and can be adapted to diagnostic applications.

• 出版日期2009-11