Lee, JR Choi, J Choi, MJ 2024-01-21T11:34:39Z 2024-01-21T11:34:39Z 2021-09-04 2001-12 0026-265X https://pubs.kist.re.kr/handle/201004/140011 The fluorescence polarization binding assay (FPBA) using fluorescein-labeled estrogen tracer is a homogeneous assay applicable to both estrogen antibody and estrogen receptor-binding assays. Two estrogen-ethylenediamine fluoresceinthiocarbamyl (E-EDF) tracers were synthesized; estrogen-6-EDF (E-6-F) derived from 6-ketoestradiol 6-(o-carboxymethyl) oxime and estrogen-17-EDF (E-17-F) was from 17 beta -estradiol 17-hemisuccinate. In both FPBAs using antibody and receptor, E-6-F tracer (Rf(365nm) = 0.58) showed a better binding response than E-17-F (Rf(365nm) = 0.70) indicating that the 17-position of estrogen seems to play an essential role as a binding site for antibody or receptor. In the optimized conditions of FPBA for E-2 using E-6-F tracer, antibody binding (K-d = 9.4 x 10(-9) M) is 50 times sensitive than receptor binding (K-d = 4.6 x 10(-8) M). Binding responses of estrogen and its related chemicals by FPBA indicate that antibody binding assay is able to screen the structural similarity of estrogen showing some response with methyltestosterone (K-i = 2.1 x 10(-5) M). On the other hand, the receptor assay is able to screen for estrogenic chemicals such as tamoxifen (K-i = 4.5 x 10(-9) M) and diethylstilbesterol (K-i = 8.1 x 10(-7) M). Therefore, E-6-F tracer is useful as a tracer for FPBA that is able to screen for chemicals structurally similar to estrogen using antibody, and that is able to screen for chemicals functionally similar to estrogen using receptor binding assay. (C) 2001 Elsevier Science B.V. All rights reserved. English ELSEVIER SCIENCE BV LIGAND-BINDING FLUOROIMMUNOASSAY METHAMPHETAMINE IMMUNOASSAY Fluorescein labeling of estrogen for application of fluorescence polarization binding assays for antibody and receptor Article 1 MICROCHEMICAL JOURNAL, v.70, no.3, pp.229 - 238 MICROCHEMICAL JOURNAL 70 3 229 238 scie scopus 000172955800011 2-s2.0-0035657438 Chemistry, Analytical Chemistry Article; Proceedings Paper LIGAND-BINDING FLUOROIMMUNOASSAY METHAMPHETAMINE IMMUNOASSAY fluorescence polarization estrogen tracer antibody receptor