Novel biosensor for the rapid measurement of estrogen based on a ligand-receptor interaction

M. Murata, M. Nakayama, H. Irie, K. Yakabe, K. Fukuma, Y. Katayama, M. Maeda

研究成果: ジャーナルへの寄稿学術誌査読

42 被引用数 (Scopus)

抄録

A bioaffinity sensor was developed aiming at the detection of estrogen. This biosensor system is based on the specific binding of estrogen to its receptor immobilized on a gold disk electrode. The recombinant DNA encoding human estrogen receptor ligand-binding domain was expressed in bacteria using the histidine-tag fusion system. The expression of the fusion protein was under control of a bacteriophage T7 promoter, and the protein was purified under native conditions by affinity chromatography, which is based on a specific interaction between a histidine-tag, located in the N-terminus of the protein, and the Ni(II) chelate adsorbent. The protein was immobilized on an Au-electrode with Ni(II)-mediated chemisorption using a histidine tag and thiol-modified iminodiacetic acid. Cyclic voltammetric measurements showed that the reversible electrochemical reaction of a ferrocyanide/ferricyanide redox couple was suppressed by the presence of estrogen in a concentration-dependent manner. It seems reasonable to suppose that the electrostatic property of the protein layer on the electrode surface was altered by complexation with estrogen. These data suggest that this biosensor is applicable to the evaluation binding activities of the chemicals toward the human estrogen receptor.

本文言語英語
ページ(範囲)387-390
ページ数4
ジャーナルanalytical sciences
17
3
DOI
出版ステータス出版済み - 2001

!!!All Science Journal Classification (ASJC) codes

  • 分析化学

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