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Bioengineering of bacterial magnetic particles and its application to estrogen receptor-ligand binding assay

Published online by Cambridge University Press:  01 February 2011

Tomoko Yoshino
Affiliation:
[email protected], Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16, Naka-cho, Koganei, 184-8588, Japan
Chihiro Kaji
Affiliation:
[email protected], Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16, Naka-cho, Koganei, 184-8588, Japan
Tadashi Matsunaga
Affiliation:
[email protected], Tokyo University of Agriculture and Technology, Department of Biotechnology, 2-24-16, Naka-cho, Koganei, 184-8588, Japan
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Abstract

Magnetic particles are used for various biomedical applications because they are easy to both handle and separate from biological samples. Nano-sized bacterial magnetic particles (BacMPs) that display the human estrogen receptor ligand binding domain (ERLBD) on their surfaces were successfully produced by the magnetotactic bacterium, Magnetospirillum magneticum AMB-1. A receptor assay for endocrine-disrupting chemicals using ERLBD-displaying BacMPs was developed. A BacMP membrane-specific protein, Mms16 or Mms13, was used as an anchor protein to localize the ERLBD on the surfaces of BacMPs. ERLBD-BacMP complexes were assayed for competitive binding of alkaline phosphatase-conjugated 17β-estradiol (ALP-E2). Inhibition curve of ALP-E2 to the powerful antagonist, tamoxifen was generated by measuring decreases in luminescence intensity that resulted from the enzymatic reaction of alkaline phosphatase. The overall simplicity of this receptor-binding assay results in a method that can be easily adapted to a high-throughput format.

Type
Research Article
Copyright
Copyright © Materials Research Society 2008

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