Barrett, The Honors College Thesis/Creative Project Collection
Barrett, The Honors College at Arizona State University proudly showcases the work of undergraduate honors students by sharing this collection exclusively with the ASU community.
Barrett accepts high performing, academically engaged undergraduate students and works with them in collaboration with all of the other academic units at Arizona State University. All Barrett students complete a thesis or creative project which is an opportunity to explore an intellectual interest and produce an original piece of scholarly research. The thesis or creative project is supervised and defended in front of a faculty committee. Students are able to engage with professors who are nationally recognized in their fields and committed to working with honors students. Completing a Barrett thesis or creative project is an opportunity for undergraduate honors students to contribute to the ASU academic community in a meaningful way.
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We confirmed that the soluble protein MHC-A2.1 could be successfully attached to the Luminex magnetic beads and detected using the primary antibody anti-GST and the detection antibody goat mAb mouse PE. The average net MFI of the attached pA2.1-bead complex was 8182. Biotinylated A2.1 MHC complexes pre-folded with β2m and FLU M1 peptide (A2.1 monomers) were also successfully attached to Luminex magnetic beads and detected with BB7.2. The average net MFI of the detected A2.1 monmer-bead complexes was 318. The protein MHC complexes were multimerized on magnetic beads to create MHC tetramers and detected with BB7.2, PE labeled monoclonal antibody, via median fluorescent intensity with the Luminex platform. Varying protein, β2 microglobulin (β2m), and peptide concentrations were tested in a number of MHC-A2.1 protein refolding trials. Different antigenic peptides and attachment methods were also tested. However, none of the MHC-A2.1 protein folding and capture trials were successful. Although MHC-A2.1 complexes and recombinant MHC molecules could be attached to Luminex magnetic beads and be detected by Luminex arrays, soluble protein A2.1 could not be successfully expressed, refolded, captured onto Luminex beads, and detected. All refolding trials resulted in a net MFI of <25. The failed refolding and capture trials of A2.1 lead to the conclusion that human cell line HeLa lysate cannot be used to properly fold MHC molecules. However, efforts to refold the complexes onto Luminex magnetic beads are ongoing. We are also using the baculovirus expression system to refold soluble A2.1 lysate onto peptide-bead complexes.
The objective of this thesis was to establish protocols and a valid experimental design for testing whether dietary mushrooms could, in fact, be protective against CVD risk. Specifically, a case-study approach was used to validate this experimental method to test white button mushrooms and their impact on blood lipid levels and the inflammatory response. This dietary study involved preparation of two soups: a placebo, broth-based soup and one with one cup of white button mushrooms per cup of soup to provide one and a half cups of soup (and mushrooms) per day to each participant. The soup was prepared in The Kitchen Café at the ASU Downtown Campus (Phoenix, AZ).
After preparing the soup, the next goal was recruitment through listserv, local advertisements, flyers, and word of mouth of participants to test the overall plan. Over fifteen people responded; however, only one candidate met the inclusion criteria of someone at high risk of developing CVD and agreed to participate in the study. The participant visited the nutrition laboratory in downtown Phoenix (550 N. 5th Street). Anthropometric data and an initial blood draw were completed, and fourteen 1.5 cup containers of mushroom soup were dispensed to the participant. After two weeks, the individual returned and the same procedures were executed to include anthropometry and blood analysis. Even though the subject did not show changes in blood markers of CVD risk (lipids and inflammatory markers), the hypothesis for the thesis that the study design would be effective was accepted. Thus, the procedure was successful and validated and will be used in the future study.