Double evaluations of chemicals using a cocktail of fused recombinant receptors.
Some chemicals have multipotential as endocrine-disrupting chemicals (EDCs). For example, some chemicals act as both estrogens and antiandrogens. Numbers of such chemicals should be evaluated from many aspects; however, labor and expenses are generally limited. We have developed two expression systems for the wild type of human estrogen receptor alpha and the wild type of human androgen receptor fused with a maltose binding protein. They are soluble and have binding activities. They showed dose-responses to natural hormones and well-known potential EDCs. After we established each assay condition for a competitive binding assay using each receptor, we found that two assay systems can be carried out simultaneously under limited and harmonized conditions. Under harmonized conditions using a cocktail of two types of receptors, we could estimate natural hormones and potential EDCs. Interference between two assay systems was not observed under these conditions. We believe that some competitive binding assays can be carried out using a cocktail of receptors at the same time if interference among different assay systems can be avoided by choosing ideal conditions.
Tsuruga Institute of Biotechnology, TOYOBO Co. Ltd., 10-24 Toyo-Cho, Tsuruga-shi, Fukui, Japan. kazuhiro_matsui@bio.toyobo.co.jp
Kazuhiro
K
Matsui
Takuya
T
Ishibashi
Masanori
M
Oka
D001667:Binding, Competitive; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D047628:Estrogen Receptor alpha; D005260:Female; D005822:Genetic Vectors; D006801:Humans; D008024:Ligands; D008297:Male; D058619:Maltose-Binding Proteins; D016297:Mutagenesis, Site-Directed; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011944:Receptors, Androgen / Q000235:genetics / Q000378:metabolism*; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis*; D014162:Transfection
D002352:Carrier Proteins; D047628:Estrogen Receptor alpha; D008024:Ligands; D058619:Maltose-Binding Proteins; D011944:Receptors, Androgen; D011960:Receptors, Estrogen; D011993:Recombinant Fusion Proteins