Honors Theses

Date of Award

Spring 5-4-2022

Document Type

Undergraduate Thesis


Chemistry and Biochemistry

First Advisor

Susan Pedigo

Second Advisor

Davita Watkins

Third Advisor

Jason Ritchie

Relational Format



Many diseases cause chronic and painful inflammation in different body systems. One of the front-line drug classes to treat such inflammation is Nonsteroidal Anti-Inflammatory Drugs (NSAIDs). Despite the benefits of oral administration of NSAIDs, there are drawbacks to their long-term usage because they can cause detrimental effects on off-target systems in the body such as the liver, kidney, or the lining of the intestinal tract. An alternative to NSAIDs is the usage of hydrogels for targeted drug delivery. Hydrogels can provide drug delivery in a specific portion of the site of inflammation, thus allowing higher doses of medication to be able to be given without the risks of damage to other body systems. Our lab has developed a Calmodulin: M13 peptide- based hydrogel system for this purpose which is the focus of this study.

The purpose of these experiments was to develop a method to modify CCLP3, the calmodulin component of our hydrogel, to create more sulfhydryl groups such that there are more attachment sites for future modification by a drug. There are three methods of modification explored in this study: the reaction of CCLP3 with Traut’s Reagent (2-iminothiolane) in an aerobic environment; the reaction of CCLP3 with Traut’s Reagent in an anaerobic environment; and, the reaction of CCLP3 with SATA (N-succinimidyl S-acetylthioacetate )in an anaerobic environment. Traut’s Reagent and SATA have unique chemistries, each with its potential upsides and downsides. This work reports the results of the modification of CCLP3 with these reagents and the subsequent analysis of the products using Ellman’s reagent.

The quantification of sulfhydryl group addition with Ellman’s Reagent concluded that each method resulted in differing levels of modification to CCLP3. There are twenty-six potential sites of modification on CCLP3, and the reagents provided total potential modification percentages of 11.7% for Traut’s Reagent in an aerobic environment, 37.4% for Traut’s Reagent in an anaerobic environment, and 19.1% for SATA in an anaerobic environment. After consulting the literature, we have determined two potential areas of improvement by increasing the molar ratio of reagent to CCLP3. In future experiments, we recommend adding a non-sulfur-containing reducing agent such as TCEP to prevent disulfide linkages while still allowing quantification of sulfhydryl groups using Ellman’s Reagent.

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Creative Commons Attribution 4.0 International License
This work is licensed under a Creative Commons Attribution 4.0 International License.



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