Full metadata
Title
Nanosensor Fabrication and Optimization for Analyzing Shelf Study
Description
In the realm of biosensors and nanotechnology, deoxyribonucleic acid (DNA) nanosensors have demonstrated tremendous potential across diverse real-world applications, from environmental monitoring to healthcare diagnostics. Fabrication of nanosensors allows assembling and designing of DNA molecules at nanoscale with high precision and versatility. Such fabricating DNA nanosensors are quite time consuming. Hence it is important to store them in batches. However synthetic DNA molecules can be prone to degradation over time, especially when exposed to various environmental factors like light, heat, or any other chemical contaminants. To address this issue, a shelf life study of DNA nanosensors using various lyoprotectant conditions was carried out to determine the long term stability of such sensors. This study involves fabrication of the dendritic, double - stranded DNA nanosensors involving five strands L1 through L5 conjugated with pHAb fluorophores via N-hydroxysuccinimide ester reaction and acetylcholinesterase (AChE) enzyme, a core component of the sensor. This sensor was originally a fluorescent ACh-selective nanosensors designed to accommodate the BTX ligand, AChE to image the ACh release in the submandibular region of the living mice to report real time quantitative endogenous ACh release triggered by electrical stimulation. AChE enzyme is a good receptor to detect acetylcholine release in the Peripheral Nervous System (PNS). The primary objective of the study was to assess DNA nanosensors with AChE, however due to the intricate interactions, non-specific binding and cost-effectiveness, the shelf life study was carried out separately. The shelf study includes observing DNA nanosensors with different disaccharide lyoprotectants like trehalose and sucrose that were analyzed under different temperature conditions: room temperature (25ºC) and at 50 ºC for different time intervals, over a week time. Also, Observing AChE with various protectants under 50 ºC with and without lyoprotectants for various time intervals like 24 hours and 48 hours. To replicate the real-world transit scenarios, the study also involves test-shipment of the samples with lyoprotectants for 2-3 days to both cross-country and local (in-state). As a result, the use of lyoprotectants, particularly trehalose, has proven to be more resilient and effective in preserving the stability and integrity of DNA nanosensors and Acetylcholinesterase (AChE) enzymes
Date Created
2023
Contributors
- Srinivasan, Nikita (Author)
- Clark, Heather A (Thesis advisor)
- Ma, Kristine Y (Committee member)
- Beeman, Scott (Committee member)
- Arizona State University (Publisher)
Topical Subject
Resource Type
Extent
65 pages
Language
eng
Copyright Statement
In Copyright
Primary Member of
Peer-reviewed
No
Open Access
No
Handle
https://hdl.handle.net/2286/R.2.N.190722
Level of coding
minimal
Cataloging Standards
Note
Partial requirement for: M.S., Arizona State University, 2023
Field of study: Biomedical Engineering
System Created
- 2023-12-14 12:48:14
System Modified
- 2023-12-14 12:48:19
- 11 months 1 week ago
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