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<p>The investigators have shown that significantly improved sensitivity can be achieved by using engineered bio-active nanoparticles. The applications of the proposed method development, if successful, will simplify pathogen detection in a variety of contexts, including food, water and medical samples.</p>
<p>The objective of the proposed research is to identify, understand, and ultimately engineer the properties of phage reporters and membranes that provide sensitive response in lateral flow assays (LFA). The hypothesis is tailoring the properties of phage nanoparticles and LFA membranes will enhance phage transport and binding and as a result reduce LFA limit of detection. The specific aims are to: (1) vary phage morphology to understand phage performance as LFA reporters; (2) identify membrane properties that enhance phage transport and adhesion onto the membrane; and (3) select phage for superior performance using directed evolution. Phage reporters of varied shape and size will be imaged with confocal microscopy as they diffuse through and bind to LFA membranes. Using high-throughput image-processing algorithms, phage orientation and dispersion, binding will be correlated to membrane properties and pore size. Directed evolution will be used to select phage with improved transport and specificity from libraries of billions of candidates. Intellectual Merit: This project integrates the expertise of the PI in imaging and confined transport with that of the co-PI in chromatographic interactions and phage engineering to develop LFAs of increased sensitivity. Phage reporters are potentially transformative as LFA reporters, enabling fast, inexpensive and ultrasensitive detection with extremely low non-specific binding. The proposed research will establish ultrasensitive LFAs as a broadly-applicable platform technology. The transport of nonspherical nanoparticles in porous media is under-investigated, and the proposed work will use phage as a new model system in for studing dispersion in complex and confined media. Finally, this proposal introduces phage evolution with separation as a novel method to engineer bacteriophage for applications in the emerging area of nanobiotechnology. Broader Impacts: Viral nanoparticle LFA technology can be readily integrated with microfluidics and smartphone-based fluorescence imaging to yield an inexpensive yet ultrasensitive point-of-use diagnostic tool. Bioseparations dominate the cost and process complexity of manufacturing of modern biopharmaceuticals; process analytical (PAT) technologies that do not require trained investigators will immediately impact this area. In addition, a field-portable analytical method may prove useful for environmental contamination, food safety monitoring, and medical diagnostics. The University of Houston is the second-most ethnically diverse research university in the US and is designated as a Hispanic-serving Institution, providing a fertile recruiting ground for broadening participation from underrepresented groups. The PIs will continue ongoing outreach efforts aimed at the local scientific community, by organizing the Texas Soft Matter Meeting for academic and industrial scientists, and at society at large, by writing and presenting popular science radio programs for The Academic Minute and Engines of our Ingenuity.</p>