Date of Graduation

5-2014

Document Type

Thesis

Degree Name

Master of Science in Biomedical Engineering (MSBME)

Degree Level

Graduate

Department

Biomedical Engineering

Advisor/Mentor

Hestekin, Christa N.

Committee Member

Wolchok, Jeffrey C.

Second Committee Member

Thallapuranam, Suresh

Keywords

Amylin; Capillary Electrophoresis; hIAPP; Protein Aggregation

Abstract

Amylin (hIAPP) aggregates have been found in 90% of patients with type II diabetes at autopsy, and are suspected to play a role in the death of islet &beta-cells1. However, this aggregation process is not well understood. Here, we explore methods that utilize capillary electrophoresis (CE) as a means to better understand amylin's aggregation process.

We examined the effects of solutions conditions: agitation, pH, salt, and temperature on amylin aggregation using Thioflavin T, dot blots, and capillary electrophoresis. Thiofalvin T was used to predict the lag time to &beta-sheet formation. Our results indicated all variables with the exception of agitation were feasible for study with CE. Capillary electrophoresis was then employed to observe the formation of oligomers with dot blots used as a confirmation technique. Overall, results showed all solutions conditions examined promote aggregation, however there was variance between time courses. Conflicting results suggest further study is needed to fully understand the observed amylin aggregation phenomena.

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