Sensors for rapid and sensitive detection of human pathogens are fundamental to biodefense screening and emergency response procedures. The availability of sensors that can additionally detect antibodies against such pathogens would indicate prior exposure, as well as aid in the development of vaccines. New, sensitive, and efficient methods of pathogen and antibody detection are being developed for such applications. Protein arrays have the power to screen samples for multiple targets simultaneously, while cell- based biosensors exploit the sensitivity of endogenous cell signaling machinery. However, these technologies remain to be fully deployed for biodefense applications. The product we propose here, the Self- Manufacturing, Assembling, and Reporting of Targets (SMART) Array, combines the power of multiplexed protein arrays with the detection sensitivity of cell-based biosensors. The SMART array offers the capability of detecting dozens to hundreds of targets in a sample simultaneously, using endogenous cell machinery to self-assemble its protein components and to self-report target binding. The SMART array thus overcomes the major limitations of protein arrays (expensive production, poor stability, and low detection sensitivity) while offering the benefits of cell-based biosensors (high sensitivity and label-free detection). The versatility of its design permits the detection of pathogens and antibodies. The first commercial product that will result from this proposal will be a SMART array capable of simultaneously detecting thirty different biodefense pathogens and antibodies. The prototype constructed in this Phase 1 proposal will focus on influenza A due to its importance as a human pathogen and the need to monitor its global spread. Each of the steps required to create the SMART array has been demonstrated to be feasible, thus mitigating the risk of its development.
The Specific Aims of this proposal are: ? SPECIFIC AIM I. Construct a Prototype SMART Array for the Detection of Pathogens and Antibodies.
SPECIFIC AIM II. Detect Pathogens and Antibodies using the SMART Array. ? ?