Date Approved

6-7-2017

Embargo Period

6-7-2017

Document Type

Thesis

Degree Name

MS Pharmaceutical Sciences

Department

Chemistry and Biochemistry

College

College of Science & Mathematics

First Advisor

Yang, Catherine

Second Advisor

Jonnalagadda, Subash

Third Advisor

Caputo, Gregory

Subject(s)

Vaccines; Food allergy

Disciplines

Medicinal-Pharmaceutical Chemistry

Abstract

Presented in this master's thesis are several studies carried out to determine the viability of several allergoid candidates utilizing the major peanut allergen Ara h 2. The Ara h 2 allergen protein appears naturally as a doublet when observed by gel electrophoresis, SDS-PAGE. Optimization of allergen purification methods successfully led to Ara h 2 purity, and the ability to standardize procedures to yield consistently pure samples. The purified allergen was chemically crosslinked with diketone derivatives selected for their abilities to react with specific amino acids accessible on the Ara h 2 protein. Chemically modified allergen samples were also evaluated using SDS-PAGE; successful protein modifications were identified by doublet band smearing or even oligomer formations such as dimers and trimers. Immunoassays were then applied to determine if epitope surface region is disrupted to indicate the diminished immune responsiveness to the modified allergen. Ara h 2 IgE specific antibodies, through western blot analysis, were used to determine antibody affinity between chemically modified and un-modified allergen proteins and to characterize any differences between the allergen samples. Results indicate that Ara h 2 may be chemically crosslinked using functional active reagents and that is possible to chemically shield IgE epitope surface regions after modification.

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