SBIR-STTR Award

The goal of this project is to develop a handheld biosensor for rapid detection of bioanalytes such as proteins, peptides, nucleic acids, toxins, viruses, bacteria and other molecular targets that represent interest for the biodefense area. We propose to build a rugged device, similar to a palm computer, equipped with a disposable sensor cartridge designed for simultaneous detection of multiple target molecules with sensitivity at the level of single molecule detection. Inexpensive sensor cartridge will have an array of printed receptor molecules and set of reagents for a one-step detection process. For many existing technologies, rate of response is slow, which makes them useless for biodefense applications that need results in several minutes or less. In contrast to currently available technologies that require multiple stages of reagent injection and washing, the envisioned biosensor system is based on a one-stage process that allows detection of molecular signatures in a matter of seconds or a few minutes. Complex samples such as whole blood, saliva, suspensions of cells, and contaminated turbid solutions can be analyzed with no or minimum sample preparation stages. This SBIR project builds upon the experience of the investigators at TIRF Technologies in developing total internal reflection fluorescence (TIRF) biosensor systems. We have built a benchtop, analytical grade, versatile TIRF biosensor instrument, which detects single biomolecules. In the proposed project we will use our previous experience to develop a handheld multianalyte biosensor with exceptionally low probability of false positive and false negative responses. The proposed technology has the potential to revolutionize many areas that are related to life sciences, including the biodefense potential of the Department of Homeland Security. In Phase I we will demonstrate the feasibility of the envisioned handheld biosensor, and in Year 1 of Phase II build its upgradeable prototype. During Phase II we will partner with academic groups to test the biosensor on 20-30 bioanalytes. We have in-house facilities for rapid prototyping and small-volume manufacturing. In Year 2 of Phase II, anticipating large demand for the envisioned handheld biosensor, we plan to partner with manufacturing companies to start large-scale production. The envisioned biosensor can find broad application for biodefense, biomedical, environmental and forensic analyses.

The overall goal of this project is to develop a handheld biosensor for rapid and sensitive detection of multiple biological analytes. The project primarily addresses the needs of first responders for detection of bioterrorism agents directly at the site of suspected attack. TIRF Technologies proposes to develop such handheld biosensor based on total internal reflection fluorescence (TIRF) combined with electrochemistry (TIRF-EC). The envisioned TIRF-EC sensor is capable of detecting multiple bioanalytes in a matter of several seconds or a few minutes. The detection limit is at the level of single molecules. Inexpensive sensor cartridge carries array of receptor molecules and contains all reagents for detection. The sensor cartridge is also equipped with a module, which performs sample preparation. To carry out the analysis, the operator inserts the cartridge, applies unpurified sample to the entrance port, and in several minutes reads the results of analysis at the handheld device. If manufactured massively, the envisioned biosensor will cost $200-400, and disposable sensor cartridges - $5-10. In Phase I, the feasibility of highly multiplexed handheld TIRF-EC biosensor was successfully demonstrated. A 4-pixel prototype of TIRF-EC sensor was built and tested. Since the electronics of 4-pixel prototype is capable of supporting up to 100 photodetectors, only several quantitative steps separate the project from building a highly multiplexed handheld biosensor. This Phase II application seeks support to develop a 64-analyte handheld biosensor based on TIRF-EC. Phase I also demonstrated the feasibility of rapid sample preparation integrated into miniature TIRF-EC sensor cartridge. In Phase II, 64-pixel TIRF-EC sensor and respective sensor cartridges will be developed, prototyped and validated in conjunction with reagentless fluorescence assays for detection DNA/RNA and protein signatures of bioterrorism agents. The envisioned handheld TIRF-EC biosensor will enable first responders with a powerful tool capable to rapidly detect multiple bioterrorism agents with minimum false positive and false negative responses. This will allow for designing efficient counter measures and avoiding unnecessary interruptions of normal life. TIRF-EC biosensor represents a flexible platform technology; it can be reconfigured for other applications. Prospective market for the envisioned handheld biosensor encompasses biomedical, forensic, food analysis, agricultural, and environmental applications