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Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry

Stuart, Orville Dean
http://rave.ohiolink.edu/etdc/view?acc_num=ucin1245823953

Abstract Details

2009, MS, University of Cincinnati, Arts and Sciences : Chemistry.
Cerebrospinal fluid (CSF) surrounds and serves as a protective media for the brain and central nervous system (CNS). This fluid remains isolated from other biological matrices in normal bodily conditions, therefore, an in depth analysis of CSF has the potential to reveal important details and malfunctions of many diseases that plague the nervous system. Because phosphorylation of a wide variety of proteins governs the activity of biological enzymes and systems, a method for the detection of 31P in proteins found in human cerebrospinal fluid by high-performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICPMS) is described. Specifically, it is of interest to compare phosphorylated proteins/peptides from patients suffering from post subarachnoid hemorrhage (SAH) arterial vasospasms against CSF from non-diseased patients. HPLC provides a way to separate many of the components of the CSF matrix, while ICPMS allows for the simultaneous detection of various elements, specifically phosphorus, at high sensitivity. Because structural elucidation is lost in the ICPMS experiment, softer ionization techniques such as Electrospray Ionization (ESI) are thus utilized for potential protein/peptide identification. Database searching software (Spectrum Mill) is then used in this study as a means of identification and validation. SEC-ICPMS confirms the presence of phosphorylated peptides within normal CSF as well as vasospastic and non-vasospastic SAH CSF samples. Non-enzymatically digested samples with ESI-MS analysis, in conjunction with Spectrum Mill database searching, indicates a variance among phosphorylated protein species across the sample batches, however, no correlations are drawn between disease types and phosphoprotein presence or absence.
Joseph Caruso, PhD (Committee Chair)
James Mack, PhD (Committee Member)
44 p.
Analytical Chemistry
ICPMS; HPLC; ESI-MS; Cerebrospinal fluid; subarachnoid hemmorhage; phosphorus

Recommended Citations

Citations

  • Stuart, O. D. (2009). Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry [Master's thesis, University of Cincinnati]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1245823953

    APA Style (7th edition)

  • Stuart, Orville. Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry. 2009. University of Cincinnati, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ucin1245823953.

    MLA Style (8th edition)

  • Stuart, Orville. "Investigation of Phosphorylated Proteins and Peptides in Human Cerebrospinal Fluid via High-Performance Liquid Chromatography Coupled to Elemental and Molecular Mass Spectrometry." Master's thesis, University of Cincinnati, 2009. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1245823953

    Chicago Manual of Style (17th edition)

ucin1245823953
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© 2009, all rights reserved.
This open access ETD is published by University of Cincinnati and OhioLINK.