| Inexpensive and robust analytical techniques for detecting molecular recognition events are in great demand in healthcare, food safety, and environmental monitoring. Despite vast research in this area, challanges remain to develop practical biomolecular platforms that, meet the rigorous demands of real-world applications. This includes maintaining low-cost devices that are sensitive and specific in complex test specimens, are stable after storage, have short assay time, and possess minimal complexity of instrumentation for readout. Nanostructured porous silicon (PSi) material has been identified as an ideal candidate towards achieving these goals and the past decade has seen diverse proof-of-principle studies developing optical-based sensing techniques.;In Part 1 of this thesis, the impact of surface chemistry and PSi morphology on detection sensitivity of target molecules is investigated. Initial proof-of-concept that PSi devices facilitate detection of protein in whole blood is demonstrated. This work highlights the importance of material stability and blocking chemistry for sensor use in real world biological samples. In addition, the intrinisic filtering capability of the 3-D PSi morphology is shown as an advantage in complex solutions, such as whole blood. Ultimately, this initial work identified a need to improve detection sensitivity of the PSI biosensor technique to facilitate clinical diagnostic use over relevant target concentration ranges.;The second part of this thesis, builds upon sensitivity challenges that are highlighted in the first part of the thesis and development of a surface-bound competitive inhibition immunoassay facilitated improved detection sensitivity of small molecular weight targets (opiates) over a relevant clinical concentration range. In addition, optimization of assay protocol addressed issues of maintaining stability of sensors after storage. Performance of the developed assay (specificity and sensitivity) was then validated in a blind clinical study that screened real patient urine samples (n=70) for opiates in collaboration with Strong Memorial Hospital Clinical Toxicology Laboratory. PSI sensor results showed improved clinical specificity over current commercial opiate immunoassay techniques and therefore, identified potential for a reduction in false-negative and false-positive screening results. Here, we demonstrate for the first time, successful clinical capability of a PSi sensor to detect opiates as a model target in real-world patient samples.;The final part of this thesis explores novel sensor designs to leverage the tunable optical properties of PSi photonic devices and facilitate colorimetric readout of molecular recognition events by the unaided eye. Such a design is ideal for uncomplicated diagnostic screening at point-of-care as no instrumentation is needed for result readout. The photonic PSi transducers were integrated with target analyte-responsive hydrogels (TRAP-gels) that upon exposure to a target solution would swell and dissolute, inducing material property changes that were optically detected by the incorporated PSi transducer. This strategy extends target detection throughout the 3-ll internal volume of the PSi, improving upon current techniques that limit detection to the surface area (2-ll) of PSi. Work to acheive this approach involved design of TRAP-gel networks, polymer synthesis and characterization techniques, and optical characterization of the hybrid hydrogel-PSi material sensor. Successful implementation of a hybrid sensor design was exhibited for a model chemical target (reducing agent), in which visual colorimetric change from red to green was observed for above-threshold exposure to the chemical target. In addition, initial proof-of-concept of an opiate responsive TRAP-gel is also demonstrated where cross-links are formed between antibody-antigen interactions and exposure to opiates induces bulk gel dissolution. |
