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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Environmental Science / Faculty of Environmental Earth Science >
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New fluorometric enzyme immunoassay for 17β-estradiol by homogeneous reaction using biotinylated estradiol
| Title: | New fluorometric enzyme immunoassay for 17β-estradiol by homogeneous reaction using biotinylated estradiol |
| Authors: | Matsumoto, Yuko Browse this author | | Kuramitz, Hideki Browse this author | | Itoh, Shinji Browse this author | | Tanaka, Shunitz Browse this author →KAKEN DB |
| Keywords: | Fluorometric enzyme immunoassay | | 17β-Estradiol | | Biotin-immobilized microtiter plate | | Solid-phase avidin–biotin binding assay |
| Issue Date: | 15-May-2006 |
| Publisher: | Elsevier |
| Journal Title: | Talanta |
| Volume: | 69 |
| Issue: | 3 |
| Start Page: | 663 |
| End Page: | 668 |
| Publisher DOI: | 10.1016/j.talanta.2005.10.040 |
| PMID: | 18970619 |
| Abstract: | A new fluorometric enzyme immunoassay for 17β-estradiol (E2) using biotinylated estradiol (BE) as a probe ligand, is described. In this method, E2 is detected indirectly by a solid-phase avidin-biotin binding assay, in which the biotin is immobilized on a microtiter plate (biotin-plate). After the competitive reaction between E2 and BE for the anti-E2 antibody in solution, the free E2 and BE are separated from the bound forms by means of ultrafiltration. The concentration of BE in the solution is determined from the reaction between the biotin immobilized on the plate and the free BE for the limited biotin binding sites of avidin conjugated with horseradish peroxidase (avidin-HRP), which is added to the solution. The enzymatic reaction of HRP was TM measured by a fluorometric analysis with the QuantaBlu Fluorogenic Peroxidase Substrate (QFPS) in order to detect of the avidin-biotin binding with a high degree of sensitivity. The detection limit and linear range for the determination of E2 were 0.12 nM and from 0.12 to 25 nM, respectively. The relative standard deviations (RSD) for the E2 assay were between 2.2 and 9.1% (n = 3). The cross-reactivity for several other estrogens was also evaluated. |
| Relation: | http://www.sciencedirect.com/science/journal/00399140 |
| Type: | article (author version) |
| URI: | http://hdl.handle.net/2115/14472 |
| Appears in Collections: | 環境科学院・地球環境科学研究院 (Graduate School of Environmental Science / Faculty of Environmental Earth Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
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Submitter: 松本 優子
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