Microfluidic device for high-throughput screening of protease specificity utilizing fluorescent resonance energy transfer

PHYS 412

Maxwell S. Bonner, maxbonner@c-gate.net1, Jonathan Fretwell2, Thomas L. Selby2, and Andre J. Gesquiere, andre@mail.ucf.edu1. (1) NanoScience Technology Center and Department of Chemistry, University of Central Florida, 12424 Research Parkway Suite 400, Orlando, FL 32826, (2) Department of Chemistry, University of Central Florida, 4000 Central Florida Blvd, Orlando, FL 32816
The challenges of high-throughput assaying are illustrated by a number of active research areas in chemical biology – assay construction, screening libraries, delivery methods, detection methods, and data analysis. However, the fabrication of microfluidic “Lab on a chip” (LOC) systems for biological diagnostics has shown promising developments for the medical sector. These devices have been applied to DNA analysis, irreversible reactions (kinetics), molecular screening and sorting. LOC has the flexibility to incorporate high-sensitivity and high-throughput screening, while utilizing small scale analysis and cost effective fabrication methods. Assembled micron scale channels (<10 μm) allow for detection down to single molecules or single cell components with nano- to picoliter sample volumes. Here, we report a fluorescent resonance energy transfer (FRET) based assay for parallel screening of protease samples at picomolar concentrations. The advantages of our LOC device include sensitivity to single protease molecules, high throughput parallel assaying with multiple targets, and low cost fabrication methods that can be scaled to volume production.