Explosive Residue Detection Using Polypeptide-Based Biosensors
Abstract
The overall goal of the proposal was to create polypeptide-based biosensors that are targeted against 2,4,6-trinitrotoluene (TNT) or its breakdown products. To identify these receptors we developed an in vitro evolution and selection (IVES) process that leads to a greatly accelerated cycle of receptor modification and diversification. During this funding period we 1) substantially optimized the IVES process by increasing the sensitivity and reducing the false positive rate of the assay; 2) developed a signal-generation system that uses fluorescence resonance energy transfer for the detection of small organic compounds; and 3) identified a TNT receptor that specifically binds TNT compared to its metabolite DNT. The EC50 for the TNT receptor in the yeast-based one-hybrid assay is ^3 M (0.7 ppm). The amount of TNT present in soil samples above a TM-62P mine buried 4" below the surface has been measured to be ^2 ppm. Thus, theoretically, a 2 ppm concentration of TNT should be detected by the TNT receptor that we developed through the IVES process.
Document Details
- Document Type
- Technical Report
- Publication Date
- Mar 07, 2005
- Accession Number
- ADA430817
Entities
People
- Binh P. Nguyen
- Danielle Kitt
- David B. Clark
- Deborah A. Lannigan
- Haoyu Qian
Organizations
- University of Virginia