Detection of hormone active chemicals using genetically engineered yeast cells and microfluidic devices with interdigitated array electrodes.
Endocrine disruptors that act like hormones in the endocrine system might have toxic effects. Therefore, it is important to develop a portable device that can detect hormone active chemicals in samples rapidly and easily. In this study, a microfluidic device was developed for the detection of hormone active chemicals using genetically engineered yeast cells. The yeast cells were used as biosensors since they were genetically engineered to respond to the presence of hormone active chemicals by synthesizing beta-galactosidase (beta-gal). For achieving further sensitivity, we incorporated interdigitated array (IDA) electrodes (width, 1.2 microm; gap, 0.8 microm) with 40 electrode fingers into the analytical chamber of the microfluidic device. The yeast cells precultured with a hormone active chemical, 17beta-estradiol (E2), were trapped from the main channel of the device to the analytical camber by electrophoresis. After trapping in the analytical chamber, we performed electrochemical detection of beta-gal induced in the yeast cells with the IDA electrodes. Actually, electrochemical detection was performed on p-aminophenol that was converted from p-aminophenyl-beta-D-galactopyranoside with beta-gal. The electrochemical signals from the yeast cells precultured with 17beta-estradiol were successfully detected with the device. Furthermore, the inhibitory effects of antagonists such as tamoxifen were also detected electrochemically by using the device. Thus, the present microfluidic device can be used for highly sensitive detection of hormone active chemicals.
Ino
Kosuke
K
Graduate School of Environmental Studies, Tohoku University, Sendai, Japan.
Kitagawa
Yusuke
Y
Watanabe
Tsuyoshi
T
Shiku
Hitoshi
H
Koide
Masahiro
M
Itayama
Tomoaki
T
Yasukawa
Tomoyuki
T
Matsue
Tomokazu
T
D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D004563:Electrochemistry / Q000295:instrumentation*; D004566:Electrodes; D004867:Equipment Design; D004958:Estradiol / Q000032:analysis*; D017930:Genes, Reporter; D005818:Genetic Engineering; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D012680:Sensitivity and Specificity; D001616:beta-Galactosidase / Q000032:analysis / Q000235:genetics
D004958:Estradiol; D001616:beta-Galactosidase