Nucleoprozyme Receptors for Biological Detection

Echo Technical, with Dr. Ellington at the University of Texas at Austin, has performed initial testing of a demonstration biosensor capable of using aptamer-based receptors (Biological Reconfigurable Interface Electronics For Classification and Analysis of Selected Elements, or BRIEFCASE). After initial testing, we found that aptamer-based biological sensors as currently developed are not sufficiently sensitive to provide detection of pathogens before wide-spread exposure and infection have already occurred, and a more aggressive solution would be required to meet the needs of Homeland Security. Dr. Ellington has devised a preliminary design for a new type of biosensor receptor (a so-called ¿nucleoprozyme¿, or NPZ) with optimal signaling characteristics. We propose to develop this new biosensor molecule that is non-reactive in a benign environment, but which spontaneously amplifies the detection of specific pathogens. This novel biosensor would produce an extremely low background signal, but in the presence of a targeted analyte signal detections rapidly and with high reliability. This new NPZ molecule and a prototype NPZ-based biosensor will be demonstrated to have performance, logistical, and cost advantages over the best available antibody-based commercial sensors, with feasibility demonstrated by the end of Phase I.The goal of Phase I is to determine the feasibility of using NPZ molecules as biosensor receptors. This will be accomplished by demonstrating the detection of nucleoprotein complexes that do not have catalytic activity, with the following five objectives to be met:1) Modify the BRIEFCASE (developed by Echo Technical) to be compatible with the requirements of NPZ receptors (i.e., design and fabricate the Mini-BRIEFCASE). 2) Select nucleic acids that complement a beta-galactosidase deletion variant. 3) Demonstrate that nucleic acids that complement a beta-galactosidase deletion variant can be used to sensitively detect sequence information. 4) Show that beta-galactosidase can generate an optical signal that can be sensitively detected in the Mini-BRIEFCASE. 5) Show that surrogate nucleic acid biosensors can be immobilized and used for sensitive target detection in the Mini-BRIEFCASE. By the completion of Phase III of this SBIR, the technology developed here promises a commercially viable long shelf-life multiplexed configurable NPZ-based biosensor prototype. This biosensor will meet several interesting market needs, including one example recently identified by HSARPA in Research Announcement 03-01, for rapid response biological identification systems. This biosensor would also be applicable to first-responder equipment for sale to state and local governments, DoD force protection applications, and medical diagnostic applications in the private sector