• Login
    View Item 
    •   Home
    • UA Graduate and Undergraduate Research
    • UA Theses and Dissertations
    • Dissertations
    • View Item
    •   Home
    • UA Graduate and Undergraduate Research
    • UA Theses and Dissertations
    • Dissertations
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Browse

    All of UA Campus RepositoryCommunitiesTitleAuthorsIssue DateSubmit DateSubjectsPublisherJournalThis CollectionTitleAuthorsIssue DateSubmit DateSubjectsPublisherJournal

    My Account

    LoginRegister

    About

    AboutUA Faculty PublicationsUA DissertationsUA Master's ThesesUA Honors ThesesUA PressUA YearbooksUA CatalogsUA Libraries

    Statistics

    Most Popular ItemsStatistics by CountryMost Popular Authors

    Mass Spectrometry: Toward Elucidating the Biosignature of Coccidioidomycosis and Insights into Surface Induced Dissociation of Biologically Relevant Carbohydrates

    • CSV
    • RefMan
    • EndNote
    • BibTex
    • RefWorks
    Thumbnail
    Name:
    azu_etd_14320_sip1_m.pdf
    Size:
    6.007Mb
    Format:
    PDF
    Download
    Author
    VanSchoiack, Andrew D.
    Issue Date
    2015
    Keywords
    Mass Spectrometry
    Surface-induced dissociation
    Chemistry
    Coccidioidomycosis
    Advisor
    Montfort, William
    Wysocki, Vicki H.
    
    Metadata
    Show full item record
    Publisher
    The University of Arizona.
    Rights
    Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
    Embargo
    Release 07-Dec-2016
    Abstract
    Mass spectrometry (MS) has proven itself to be indispensable for the analysis of biomolecules and molecular systems. This research has three goals: (1) expand on prior work toward the discovery of novel diagnostic targets for Valley Fever, (2) evaluate current mass spectrometry based proteomics for the discovery of non-host protein in complex host biological samples, and (3) investigate the potential for two gas phase techniques, surface induced dissociation, and ion mobility for the analysis of carbohydrate based molecules. Mass spectrometry has allowed for great advances in the identification of proteins in biological samples through implementing liquid chromatography tandem mass spectrometry and bioinformatics techniques known as proteomics. Proteomics techniques were used to elucidate a portion of the biosignature of Valley Fever (VF), a disease of great importance in the arid regions of the western United States. Current diagnostics for this fungal lung disease are remarkably unreliable which creates a need for an unfailing diagnostic method. Using a new generation of instrumentation along with directed methods, four previously discovered VF marker proteins were evaluated for their presence in mouse plasma, lung homogenate and bronchoalveolar lavage fluid samples. Due to inconclusive data, discovery proteomics approaches were then used to identify possible diagnostic targets in both human and mouse bronchoalveolar lavage fluid. In human bronchoalveolar lavage fluid, one potential target was discovered in five out of eight VF positive samples, and two further identifications of VF in negative samples. Mouse bronchoalveolar lavage fluid also showed the presence of this protein. Multiple-reaction monitoring based validation, using two-dimensional online separations for the presence of either the newly discovered protein or the four previously discovered proteins, was inconclusive. Emerging from the difficulties observed by the author and colleagues in identifying infectious agent proteins in complex host biological samples, an investigation of the feasibility of undertaking such endeavors was performed. One of the main complications thwarting the discovery of infectious agent proteins is the dynamic range of protein concentration in the host biological sample. This issue was resolved by using commercially available mass spectrometry and a two-dimensional liquid chromatographic separations platform. This enhanced separation combined with cost-effective protein normalization techniques, identified non-host proteins with good sequence coverage and spectral counts. Combining antibody-based depletion of highly abundant plasma proteins in bronchoalveolar lavage fluid, with at least a three fraction sample analysis enabled detection of a low abundant non-host protein (2pmol in 50μg host protein) with high sequence coverage. Glycosylation, an abundant post-translational modification of protein composed of carbohydrate oligomers may hold within its structure more biologically relevant information than the DNA that encoded the protein on which the glycan resides. The analysis of glycosylation plays a critical role in understanding biology. Carbohydrate based moieties pose many distinct challenges to their analysis; two of which are isobaric fundamental units and complex branching chemistry. Mass spectrometry provides a way of overcoming some of these challenges. To examine the complex biomolecules, a gas phase ion separation technique, known as ion mobility, and a non-traditional ion activation technique, surface-induced dissociation, were used. Surface-induced dissociation provides analogous fragmentation patterns to those generated via collision-induced dissociation (CID); however, much more extensive fragmentation can be achieved in a single tandem MS experiment. Using the gas-phase separations power of ion mobility showed that multiple conformations were adopted by relatively simple oligosaccharides. Ion mobility was also successfully used to determine fragment ion lineage of isobaric fragment ions, through inline separation between two differential fragmentation experiments.
    Type
    text
    Electronic Dissertation
    Degree Name
    Ph.D.
    Degree Level
    doctoral
    Degree Program
    Graduate College
    Chemistry
    Degree Grantor
    University of Arizona
    Collections
    Dissertations

    entitlement

     
    The University of Arizona Libraries | 1510 E. University Blvd. | Tucson, AZ 85721-0055
    Tel 520-621-6442 | repository@u.library.arizona.edu
    DSpace software copyright © 2002-2017  DuraSpace
    Quick Guide | Contact Us | Send Feedback
    Open Repository is a service operated by 
    Atmire NV
     

    Export search results

    The export option will allow you to export the current search results of the entered query to a file. Different formats are available for download. To export the items, click on the button corresponding with the preferred download format.

    By default, clicking on the export buttons will result in a download of the allowed maximum amount of items.

    To select a subset of the search results, click "Selective Export" button and make a selection of the items you want to export. The amount of items that can be exported at once is similarly restricted as the full export.

    After making a selection, click one of the export format buttons. The amount of items that will be exported is indicated in the bubble next to export format.