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Estrogen receptor binding assay of chemicals with a surface plasmon resonance biosensor
- Source :
- The Journal of steroid biochemistry and molecular biology. 81(1)
- Publication Year :
- 2002
-
Abstract
- We have developed a simple assay method for the evaluation of estrogen receptor (ER) binding capacity of chemicals without the use of radio- or fluorescence-labeled compounds. We used the solution competition assay by the BIACORE biosensor, a surface plasmon resonance biosensor, with estradiol as a ligand, human recombinant ER(alpha) (hrER(alpha)) as a high molecular weight (hmw) interactant and test chemicals as analytes. For the ligand, aminated estradiol with a spacer molecule (E2-17PeNH) was synthesized and immobilized on a carboxymethyl dextran-coated sensor chip by the amine coupling method. The injection of the hmw interactant hrER(alpha) to the biosensor raised the sensorgram, indicating its binding to the ligand E2-17PeNH. The binding of test chemicals to hrERalpha was determined as a reduction in the hrER(alpha) binding to E2-17PeNH. The dissociation constant for the binding to hrER(alpha) was calculated for estrone (4.29 x 10(-9)M), estradiol (4.04 x 10(-10)M), estriol (8.35 x 10(-10)M), tamoxifen (2.16 x 10(-8)M), diethylstilbestrol (1.46 x 10(-10)M), bisphenol A (1.35 x 10(-6)M) and 4-nonylphenol (7.49 x 10(-6)M), by plotting the data according to an equation based on mass action law. This method can also be used as a high throughput screening method.
- Subjects :
- Bisphenol A
Endocrinology, Diabetes and Metabolism
Clinical Biochemistry
Analytical chemistry
Biosensing Techniques
Biochemistry
chemistry.chemical_compound
Endocrinology
Humans
Surface plasmon resonance
Molecular Biology
Chromatography
Estrogen receptor binding
Chemistry
Estrogen Receptor alpha
Cell Biology
Surface Plasmon Resonance
Ligand (biochemistry)
Recombinant Proteins
Dissociation constant
Receptors, Estrogen
Molecular Medicine
Amine gas treating
Biosensor
Estrogen receptor alpha
Protein Binding
Subjects
Details
- ISSN :
- 09600760
- Volume :
- 81
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- The Journal of steroid biochemistry and molecular biology
- Accession number :
- edsair.doi.dedup.....3be362bd82c24b34702b02f89118a7d4