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Bioengineering Seminar Series: Kim Sapsford
Friday, September 17, 2010
11:00 a.m.-12:00 p.m.
Room 2108, Chemical and Nuclear Engineering Bldg.
For More Information:
Professor Peter Kofinas

Energy Transfer-Based Biosensing of Protease Activity: Potential Use in Portable Screening Applications

Kim Sapsford
Research Staff Fellow
Division of Biology
Center for Devices and Radiological Health
U.S. Food and Drug Administration

A number of Förster Resonance Energy Transfer (FRET) assays targeting proteases, including botulinum neurotoxin A (BoTN-A) and trypsin, are commercially available and offer the potential to be used as simple portable assays for various screening applications. In order to measure such FRET-based assays we have designed a portable fluorescence-based detector platform which incorporates spatial illumination of microfabricated chip devices using electroluminescence (EL) semiconductor strips. The EL excitation was combined with charge-coupled device (CCD) detector (EL-CCD) resulting in a simple portable fluorescence detector system. The resulting platform consists of two modules; (1) the detection module which houses the CCD camera and emission filters and (2) the excitation and sample module, containing the EL strip, the excitation filter and the sample chip. In later studies, the use of light emitting diode (LED) strips, to improve the illumination intensity, was investigated and directly compared to the EL surface illumination. The system was optimized and tested using a commercial FRET assay for botulinum neurotoxin A (BoTN-A), with measured limits of detection for the specific light chain A (LcA) derivative between 0.625-1.25 nM (31-62 ng/ml). Studies have also been extended to investigate the EL-CCD platform’s potential for incorporating semiconductor quantum dot (QD)-based FRET protease sensors. Results suggest that the unique photophysical properties of the QDs combined with the reduced-scale nature of the microfabricated devices make them ideal for a variety of sensitive applications including point-of-care diagnostics, screening and global healthcare applications. In addition the EL-CCD platforms ability to be used in colorimetric-based ELISA assays was demonstrated with miniature ELISA chips, reducing the over all assay fluid consumption, compared to standard 96-well plate ELISAs, while maintaining sensitivity.

This Event is For: Graduate • Faculty • Post-Docs

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