Biodiagnostic Approaches to Human Profiling Through Nanomaterial Indicators

Innovative claims for the proposed research: Our goal is to develop the next generation of bioassays by advancing nanotechnology-based components that can be utilized in intelligent and portable lab-on-a-chip technologies. These assays will be based on spherical nucleic acid-gold nanoparticle conjugates (SNA-AuNPs)1,2 and surface-enhanced Raman scattering (SERS) nanosheets.3 This approach has significant advantages over state-of-the-art methods because it avoids enzymatic amplification steps, is highly scaleable, and has a simple readout mechanism based on the optical properties of metal nanoparticles. A terrorist working to synthesize, manufacture, and use anthrax (as proposed in our threat hypothesis) will exhibit a unique set of biological signatures in their blood and sweat that can be detected with high specificity and sensitivity using these assays. Our threat hypothesis will include biomarkers, such as bacterial DNA, microRNA (miRNA), and proteins used by the bacterium, chemicals used in the production of anthrax (i.e., sporicides), and spores. Taken together these ‘signatures’ provide an important balance between agent and environmental exposure defining an individual’s involvement in the production, handling, and transport of anthrax. This proposal covering 24 months (phase 1) will put us in a position to identify a miRNA profile for anthrax exposure and develop SNA-AuNPs and SERS nanosheet assays towards a comprehensive anthrax sensor. Summary of products, technology, and deliverables of the proposed research results: We will develop: 1) A test plan for our bioassays (a detailed protocol for developing the bioassays and biomarkers of interest); 2) A threat hypothesis model, detailing the rational for our threat hypothesis; 3) The dose-response curves of our optimized assays; 4) A bioinformatics analysis of miRNA targets, demonstrating our ability to identity miRNA targets from human blood; 5) A list of biomarkers; 6) A revised test plan for our bioassays (curves, protocols, and LoDs); 7) A bioassay waypoints report; 8) Duplicate samples; 9) A final report.