Date of Graduation

5-2022

Document Type

Thesis

Degree Name

Bachelor of Science

Degree Level

Undergraduate

Department

Chemistry & Biochemistry

Advisor/Mentor

Kumar, TKS

Committee Member/Reader

Adams, Paul

Committee Member/Second Reader

Ceballos, Ruben Michael

Committee Member/Third Reader

Davidson, Fiona

Abstract

Chronic wounds pose a major problem in the United States with an estimate of twenty-five million dollars a year spent on associated treatments. Growth factors can be used as a potential treatment for chronic wounds since they promote cell proliferation and angiogenesis. This study employs one specific growth factor, fibroblast growth factor 2 (FGF2) so that it could potentially be used in future treatment. Wild-type FGF2 is thermally unstable, and it has a mean elimination time of 7.6 hours. This study attempted to improve upon its stability through a mutation on the heparin binding loop. The mutation performed was K134E. This mutation decreased the number of electrostatic interactions present on the heparin-binding loop.

FGF2 was first be attained through the large-scale expression of glutathione-S- transferase-fibroblast growth factor 2 (GST-FGF2). GST-tag was then cleaved from GST-FGF2 by thrombin digestion. The stability and structure were studied through differential scanning calorimetry, circular dichroism, and florescence. This process is a low-cost method since GST-FGF2 was purified from an E. coli culture containing the mutant plasmid

Even though both wild-type and mutant protein were successfully purified at least once, there was not enough protein purified for neither to conduct all characterization studies. A purification procedure was established, and it is possible that the mutant has greater thermal stability than the wild-type but further studies are still needed to make a definitive conclusion.

Keywords

Chronic wounds, cell proliferation, fibroblast growth, heparin-binding loop

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