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Phage Binding for Continuous Anthrax Spore Detection

$290,000R21FY2004AINIH

Auburn University At Auburn, Auburn University AL

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Abstract

DESCRIPTION (provided by investigator): Spores of Bacillus anthracis (BAS) poses a major bioterrorism threat because of their extreme potency and availability. There is an urgent need to rapidly detect BAS for efficient treatment of anthrax. The ultimate goal of this application is the development of a device that could be mounted to a SpinCon (air-to-liquid) concentrator for the continuous monitoring, detection and alarm of the presence of airborne anthrax spores. Fluid from the SpinCon would be directed across the surface of the Auburn University-designed sensor, which could continuously monitor for the presence of anthrax spores. Detection would occur by the binding of the anthrax spores to phage-derived probes (BAS probes) specifically designed to target these spores. BAS-probes will be selected from the billion-clone phage libraries expressing foreign random peptides on all 4,000 copies of the viral major coat protein (landscape libraries), and will be improved using mutagenesis and affinity maturation. The phage particles demonstrating best binding to BAS in Enzyme Linked Immuno Sorbent Assay (ELISA) will be remodeled to eliminate their Escherichia coil-binding domains, and will be immobilized onto the surface of either a miniaturized plasmon surface resonance or acoustic wave resonator to produce a signal indicating the detection of anthrax spores. Specificity and selectivity of detection of BAS will be studied using BAS mixed with unrelated spores, proteins and other biopolymers. Parallel experiments will also be conducted with antibody-derived probes for comparison evaluation of specificity, selectivity and longevity of the sensors.

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