Theses and Dissertations
This analysis explores what the time needed to harden, and time needed to degrade is of a PLGA bead, as well as whether the size of the needle injecting the bead and the addition of a drug (Vismodegib) may affect these variables. Polymer degradation and hardening are critical to understand for the polymer’s use in clinical settings, as these factors help determine the patients’ and healthcare providers’ use of the drug and estimated treatment time. Based on the literature, it is expected that the natural logarithmic polymer mass degradation forms a linear relationship to time. Polymer hardening was tested by taking video recordings of gelatin plates as they are injected with microneedles and performing RGB analysis on the polymer “beads” created. Our results for the polymer degradation experiments showed that the polymer hardened for all solutions and trials within approximately 1 minute, presenting a small amount of time in which the patient would have to remain motionless in the affected area. Both polymer bead size and drug concentration may have had a modest impact on the hardening time experiments, while bead size may affect the time required for the polymer to degrade. Based on the results, the polymer degradation is expected to last multiple weeks, which may allow for the polymer to be used as a long-term drug delivery system in treatment of basal cell carcinoma.
The use of enzyme-catalyst interfaces is underexplored in the field of biocatalysis, particularly in studies on enabling novel reactivity of enzymes. For this thesis, the HaloTag® protein tagging platform was proposed as a bioconjugation method for a pinacol coupling reaction using lipases, as a model for novel reactivities proceeding via ketyl radical intermediates and hydrogen-bonding-facilitated redox attenuation. After an initial lipase screening of 9 lipases, one lipase (Candida rugosa) was found to perform the pinacol coupling of p-anisaldehyde under standard conditions (fluorescein and 530nm light, 3% yield). Based on a retrosynthetic analysis for the photocatalyst-incorporated HaloTag® linker, the intermediates haloamine 1 and aldehyde 6 were synthesized. Further experiments are underway or planned to complete linker synthesis and conduct pinacol coupling experiments with a bioconjugated system. This project underscores the promising biocatalytic promiscuity of lipases for performing reactions proceeding through ketyl radical intermediates, as well as the underdeveloped potential of incorporating bioengineering principles like bioconjugation into biocatalysis to overcome kinetic barriers to electron transfer and optimize biocatalytic reactions.