Date of Graduation

12-2015

Document Type

Thesis

Degree Name

Master of Science in Chemical Engineering (MSChE)

Degree Level

Graduate

Department

Chemical Engineering

Advisor

Christa Hestekin

Committee Member

Robert Beitle

Second Committee Member

Josh Sakon

Third Committee Member

Shannon Servoss

Abstract

According to recent literature, it is believed that the oligomeric form of amyloid beta (Aβ) is the leading cause of Alzheimer’s disease (1; 8; 10; 12-18). Additionally, recent studies have eluded to the impact of Alzheimer’s disease (AD) both economically and socially in today’s society where an increase of about 71% of AD related deaths were recorded between 2000 and 2013 (7). Since the oligomeric forms of Aβ vary in size, shape and some believe conformation, it is vital to utilize a separation technique, such as capilllary electrophoresis (CE) to further understand Aβ aggregation. By understanding Aβ aggregation, treatment of AD or preventive care measurements could be additionally developed.

Therefore in this study, field amplified sample stacking (FASS) technique on the CE was utilized to provide higher resolution in oligomeric Aβ1-42 detection without causing significant changes to the aggregation. It was observed that the FASS technique provided smaller peak widths and increased peak heights on the CE compared to the non-FASS conditions. Furthermore by conducting thioflavin-t (ThT) assays, it was observed changing the buffer concentrations in accordance to the FASS technique conditions did not effect the overall aggregation. Thioflavin-t (ThT) assays were also conducted in order to determine an agitation rate where the oligomers of Aβ1-42 were observed on the CE.

The oligomeric species observed were believed to be less than or equal to 100 kDa. Additionally, Congo red and Orange G inhibition were conducted to confirm oligomeric Aβ1-42 species were observed on the CE. Both inhibition studies alongside TEM imaging proved the aggregates observed on the CE in the 27 hour aggregation were smaller than proto-fibrils. Future work on natural compound inhibition studies using CE are recommended to see how those inhibitors target Aβ1-42 species that are less than or equal to 100 kDa.

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