Signal Amplification of Bioassay Using Zinc Nanomaterials
AuthorCowles, Chad Lewis
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An emerging trend in the analytical detection sciences is the employment of nanomaterials for bioassay signal transduction to identify analytes critical to public health. These nanomaterials have been specifically investigated for applications which require identification of trace levels of cells, proteins, or other molecules that can have broad ranging impacts to human health in fields such as clinical diagnostics, environmental monitoring, food and drink control, and the prevention of bioterrorism. Oftentimes these nanoparticle-based signal transduction or amplification approaches offer distinct advantages over conventional methods such as increased sensitivity, rapidity, or stability. The biological application of nanoparticles however, does suffer from drawbacks that have limited more widespread adoption of these techniques. Some of these drawbacks are, high cost and toxicity, arduous synthesis methods, functionalization and bioconjugation challenges, and laboratory disposal and environmental hazard issues, all of which have impeded the progression of this technology in some way or another. This work aims at developing novel techniques that offer solutions to a number of these hurdles through the development of new nanoparticle-based signal transduction approaches and the description of a previously undescribed nanomaterial.Zinc-based nanomaterials offer the opportunity to overcome some of the limitations that are encountered when other nanomaterials are employed for bioassay signal transduction. On the other hand, the biological application of zinc nanomaterials has been difficult because in general their fluorescence is in the blue range and the reported quantum yields are usually too low for highly sensitive applications. The advantages of using zinc nanomaterials for biological applications, such as reduced toxicity, simple synthesis, low cost, and straightforward functionalization strategies contribute to the research interest in their application as bioassay signal tranducers. To overcome the limitations associated with zinc-based nanomaterials, a novel signal transduction approach was developed that relies on zinc ion release from nanoparticle labels during an immunoassay. The development of an innovative method for zinc ion detection and the description of a previously undescribed zinc-based nanomaterial are also described in this work. There are three major contributions to science in this work: (1) The development of an original and innovative signal transduction approach for immunoassays that adopts fluorescence detection of zinc ions released from ZnS nanoparticle labels; (2) The discovery and development of dual signal amplification for immunoassay signal transduction using ion release and subsequent activation of a zinc dependent metallozyme; (3) The synthesis and characterization of a novel zinc-based nanomaterial and its biosensing application using both single and dual signal amplification strategies.