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Date of Award
Spring 2023
Document Type
Masters Thesis
Degree Name
Master of Science (MS)
Department
Chemistry
First Advisor
Murphy, Amanda R.
Second Advisor
Vyvyan, James R.
Third Advisor
Amacher, Jeanine
Abstract
The use of silk microparticles (µPs) as drug delivery devices has gained attention due to slow degradation properties, mild preparation conditions, and advantageous biocompatibility. However, little research has been done on where these particles go once injected. To expand these studies, the goal for this work is to create bioluminescent silk µPs that can be tracked in vivo. Here several methods are demonstrated for preparing bioluminescent silk µPs containing Nanoluciferase (NLuc) and/or its substrate furimazine (FZ). In this study, silk µPs were formed using a salting-out procedure. The first method involved non-specific adsorption of NLuc and non-specific adsorption of FZ to separate silk µPs. Light output produced by introducing these two silk µPs sample types to each other was monitored over time in buffer as well as in the presence of other proteins. This study found that stable light emission could be achieved in all scenarios tested including when NLuc was absorbed to silk µPs and then exposed to FZ in solution, when FZ was absorbed to silk µPs and then exposed to NLuc in solution, and when silk µPs containing NLuc and silk µPs containing FZ were mixed in buffer. Both types of silk µPs were also found to stabilize light emission in the presence of other proteins. For the next method, tyrosine residues of silk µPs were chemically modified to contain an azide functional group. The µPs were then soaked in dibenzocyclooctyne(DBCO)-linked NLuc solution. This resulted in the covalent linkage of silk µPs to NLuc via strain-promoted alkyne-azide cycloaddition. Light output produced by these luminescent silk µPs in the presence of FZ was monitored over extended periods of time. These two methods were compared to determine if covalently attaching the NLuc enzyme to silk µPs results in a more stable light output. This study found that both modified and unmodified a stable light output, silk µPs that had been soaked in NLuc DBCO produced indicating that covalent attachment of the NLuc enzyme to silk µPs was not necessary. The future role these BL silk µPs could play in drug delivery and cancer imaging is also discussed.
Type
Text
Keywords
silk microparticles, bioluminescence, Nanoluciferase, furimazine, drug delivery devices
Publisher
Western Washington University
OCLC Number
1381160753
Subject – LCSH
Bioluminescence; Silk; Drug delivery devices; Cancer--Imaging
Format
application/pdf
Genre/Form
masters theses
Language
English
Rights
Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this document for commercial purposes, or for financial gain, shall not be allowed without the author’s written permission.
Recommended Citation
Berg, Monique, "Development of Silk Microparticles Capable of Bioluminescence" (2023). WWU Graduate School Collection. 1198.
https://cedar.wwu.edu/wwuet/1198