Dissertation
DEVELOPMENT OF RAPID AND EFFECTIVE NANOPARTICLE-AMPLIFIED IMMUNOASSAYS TO DETECT TARGET ANALYTES
Washington State University
Doctor of Philosophy (PhD), Washington State University
01/2022
DOI:
https://doi.org/10.7273/000004533
Handle:
https://hdl.handle.net/2376/122415
Abstract
In this research, nanoparticle (NP)-amplified immunoassays were developed to detect two types of target analytes—pesticides and a protein—rapidly and effectively. These analytes are useful in developing NP-based techniques because their varied molecular weights require different ELISA methods. This dissertation contains results for three studies on mesoporous core-shell peroxidase-like Pd@Pt NPs, which can replace enzymes in ELISA-like formats. Morphology of the NPs reveal a high surface area fractal pattern, which helps explain its high catalytic activity. A competitive Pd@Pt NP-linked immunosorbent assay (NLISA) was successfully developed to detect herbicides atrazine in spiked water samples with a limit of detection (LOD) of 0.5 ng/mL and a linear range of 0.1 – 500 ng/mL. In a second study, competitive NLISAs were successfully applied to detect atrazine and acetochlor in spiked juice, fruit, and vegetable samples; results were compared to those for HPLC and a Pd@Pt NP-amplified dual-lateral flow differential pulse voltammetry immunoassay. In contrast to HPLC, the two immunoassays required simple or no sample preparation steps. Results for both immunoassays compare well to each other revealing good recoveries for spiked samples in the 87.0 – 114% range. The colorimetric catalytic intensity of Pd@Pt NPs was confirmed showing a deep blue color increasing in intensity with higher concentrations of NPs. Synthesis of antibody conjugation to NPs (Ab-NPs) was confirmed via zeta potential analysis without losing catalytic activity. Pd@Pt NP usage is extended to a macromolecule collagen type II (COL II) containing more than one epitope. The direct sandwich NLISA was developed to detect COL II in engineered tissue samples with an LOD of 0.82 ng/mL and a linear range of 3 ng/mL – 50 μg/mL. Use of NPs shows thermal stability over a 4 – 65 °C range and stability over a 7 – 9 pH range with an optimal pH of 8.5. The NP-linked immunosorbent assays have the potential for detecting multiple targets, such as pesticides and other small molecules and proteins and other macromolecules, in various fields including food and agricultural monitoring, medical diagnostics and tissue engineering.
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Details
- Title
- DEVELOPMENT OF RAPID AND EFFECTIVE NANOPARTICLE-AMPLIFIED IMMUNOASSAYS TO DETECT TARGET ANALYTES
- Creators
- Eunice Y Kwon
- Contributors
- Bernard J. Van Wie (Advisor)Dan Du (Committee Member)Cornelius F. Ivory (Committee Member)
- Awarding Institution
- Washington State University
- Academic Unit
- Voiland College of Engineering and Architecture
- Theses and Dissertations
- Doctor of Philosophy (PhD), Washington State University
- Publisher
- Washington State University
- Number of pages
- 206
- Identifiers
- OCLC#: 1363848354; 99900882927801842
- Language
- English
- Resource Type
- Dissertation