EAGER: Enhancing Biosensor Response Rates through Nanostructures with Analyte Delivery Functions

Rapid detection of biological molecules helps protect public health, monitor the environment, and enable timely medical decisions. Many biosensors operate slowly because target molecules must diffuse a long distance to reach the sensing surface. The delay increases testing costs and reduces sensitivity. This project will build a nanostructure that guides target molecules toward a sensing surface. The structure will have flexible DNA “tentacles” that attract target molecules and concentrate them near the surface. The result will be faster detection and improved sensitivity. The new technology will improve pathogen monitoring, point-of-care diagnostics, and rapid screening for genetic mutations. The project will train undergraduate and graduate students in biotechnology and biosensor engineering. The project will strengthen the biotechnology workforce and enhance health, environmental safety, and economic sustainability. This project will develop and validate a DNA-based nanostructure, termed the DNA Cephalopod, to overcome mass transfer limitations in biosensing and enhance analyte binding kinetics. The hypothesis is that analyte-binding fragments attached to tentacle strands will preconcentrate target molecules near the sensing element, thereby accelerating response rates and improving limits of detection (LOD). Two objectives will be pursued. Objective 1 will focus on designing and optimizing the DNA cephalopod based on split light-up aptamer sensors for homogeneous detection of nucleic acids. Sensor performance will be evaluated by measuring response kinetics, LOD, and single-nucleotide variant discrimination using model pathogen-derived nucleic acid targets. Objective 2 will extend the cephalopod strategy to heterogeneous sensing formats by immobilizing optimized nanostructures on silica microparticles to assess mitigation of diffusion limitations in surface-based detection. Sensor response time, signal-to-background ratio, and reproducibility will be evaluated across particle sizes and assay conditions. The expected outcomes include at least a tenfold improvement in sensor response rates and sensitivity relative to conventional hybridization sensors. The project will establish a generalizable design principle for enhancing molecular transport in biosensors and provide a foundation for integration into fluorescence, electrochemical, and optical sensing platforms. These advances will enable faster, more sensitive, and cost-effective biosensing for environmental monitoring, medical diagnostics, and biotechnology applications. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.