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Surface plasmon resonance-based immunoassay for 17β-estradiol and its application to the measurement of estrogen receptor-binding activity

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Abstract

A rapid and simple surface plasmon resonance (SPR)-based immunoassay for detection of 17β-estradiol was developed. The assay was designed as an inhibitive format, in which 17β-estradiol–BSA conjugates are immobilized on an SPR sensor chip and the binding of antibody to the chip is measured. The binding was inhibited by 17β-estradiol in the concentration range 0.468 to 21.4 nmol L−1 with an IC50 value of 2.29±0.10 nmol L−1. Although not as sensitive as traditional radioimmunoassay (RIA) and enzyme-linked immunoassay (ELISA), this method requires no separation and washing after addition of the antibody, steps which are relatively time-consuming. Estrogen receptor (ER)-binding was then investigated using this SPR immunoassay for the determination of the amount of unbound 17β-estradiol after competition with test compounds for the ER-binding. Inhibition of the binding of 17β-estradiol to ER by diethylstilbestrol (DES) was successfully measured by injecting the reaction mixture into the SPR sensor after addition of the antibody. This binding assay requires no separation of unbound 17β-estradiol from the mixture and no radioisotope- or fluorescence-labeling of 17β-estradiol. These results show the potential usefulness of the SPR sensor both detecting 17β-estradiol and evaluating the ER-binding activity of xenoestrogens such as DES in a single assay system.

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Acknowledgements

We thank Mr Yoshihiro Soya of Toyobo Co., Ltd. for helpful discussions.

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Authors and Affiliations

  1. Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan

    Masahiro Miyashita, Takahiro Shimada & Hisashi Miyagawa

  2. CREST, Japan Science and Technology Agency, Saitama, 332-0012, Japan

    Masahiro Miyashita, Takahiro Shimada & Miki Akamatsu

  3. Division of Environmental Science and Technology, Graduate School of Agriculture, Kyoto University, Kyoto, 606-8502, Japan

    Miki Akamatsu

Authors
  1. Masahiro Miyashita
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  2. Takahiro Shimada
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  3. Hisashi Miyagawa
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  4. Miki Akamatsu
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Correspondence to Masahiro Miyashita.

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Miyashita, M., Shimada, T., Miyagawa, H. et al. Surface plasmon resonance-based immunoassay for 17β-estradiol and its application to the measurement of estrogen receptor-binding activity. Anal Bioanal Chem 381, 667–673 (2005). https://doi.org/10.1007/s00216-004-2952-z

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  • DOI: https://doi.org/10.1007/s00216-004-2952-z

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