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Madou, Mark
BIO-MEMS Lab
Research Interests
  • Medical Diagnostics
  • Micro-Battery Development
  • Drug Delivery Systems

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Whole-cell-reporter-gene-based biosensing systems on a compact disk microfluidics platform

Cited 28 times inthomson ciCited 28 times inthomson ci
Title
Whole-cell-reporter-gene-based biosensing systems on a compact disk microfluidics platform
Author
Rothert, ADeo, SKMillner, LPuckett, LGMadou, MarkDaunert, S
Keywords
Arsenic; Compact disc microfluidics platform; Reporter gene assay
Issue Date
2005-07
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
Citation
ANALYTICAL BIOCHEMISTRY, v.342, no.1, pp.11 - 19
Abstract
Biosensing systems such as reporter-gene-based whole-cell assays are increasingly finding applications in biological and environmental screening. A whole-cell approach to such analyses can provide valuable information about the bioavailable level of a compound of interest. These biosensing systems rely on the molecular recognition of a specific analyte by a regulatory protein and, therefore, can detect low levels of the target analyte. In this study, Escherichia coli cells containing plasmid pSD10 were engineered to sense the model target analytes arsenite and antimonite, the target analytes in this study. The biosensing system takes advantage of the recognition of the regulatory protein, ArsR, for arsenite and antimonite to produce the reporter protein, which in this case is GFPuv. The fluorescence emitted by the GFPuv in the cells can be directly related to the concentration of the analyte in the cell, making this biosensing system useful in the detection of arsenite and/or antimonite in a variety of samples. Miniaturization of biosensing systems can further enhance their utility by decreasing reagent consumption and analysis time and by allowing for the high-throughput screening of samples. To that end, we employed a microcentrifugal microfluidics platform that has low power, space, and reagent requirements, increased speed of detection, and the potential for portability. Herein, we demonstrate for the first time the adaptation of a whole-cell sensing system to a microcentrifugal microfluidics platform. Moreover, we were able to detect our target analytes in a rapid and sensitive manner compared to conventional sensing methods
URI
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DOI
10.1016/j.ab.2004.10.048
ISSN
0003-2697
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BME_Journal Papers
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