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dc.contributor.authorBotelho, Julianne Cook
dc.description.abstractHerein we explore the applications of mass spectrometry on three different biological samples, mosquito cuticles, human blood serum, and porcine mucin. A proteomics analysis of the cast cuticles of Anopheles gambiae was performed. From this analysis 259 peptides from putative cuticular proteins were matched to 119 proteins in 69 protein families. Authentic cuticular proteins previously reported were identified following extraction, isolation and sequencing of individual proteins isolated from manually cleaned cuticles. The proteomics approach allowed for the identification of more proteins, in a high throughput manner, then previously reported. Furthermore, several of the identified proteins are reported as authentic Anopheles gambiae cuticle proteins for the first time. A glycoproteomics approach using Phaseolus Vuglaris Leukoagglutinin lectin affinity chromatography is described. This analysis allows for identification of low abundance complex type glycoproteins that carry a ²6 -N-Acetylglucosamine (GlcNAc) branch from the blood serum of patients with benign and malignant ovarian tumors. From this study 53 proteins were identified in the serum of patients with malignant tumors, 44 in patients with benign tumors, and 34 in a control sample of pooled male serum. This accounted for a total of 63 unique proteins from the 9 serum samples. From this analysis, 10 proteins were identified that with further investigation could serve as potential biomarker candidates. Nine of these proteins were identified only in the serum of patients with malignant tumors while an additional protein was observed to have a higher abundance in malignant serum based on relative quantification by normalized spectral counts. A quantitative isobaric labeling (QUIBL) technique for glycomics was validated for O-linked glycans. QUIBL, which is based on permethylation of glycans with isobaric labels, proved to be applicable for O-linked glycans based on the analysis of fetuin on a high resolution mass spectrometer. The reproducibility and accuracy of QUIBL for O-linked glycans was confirmed over 2 orders of magnitude and its application to a more complex sample, porcine mucin, was verified in this study
dc.subjectMass Spectrometry
dc.subjectMosquito Cuticular Proteins
dc.subjectOvarian Cancer
dc.subjectIsobaric labeling
dc.subjectStable isotope labeling
dc.subjectQuantitation O-linked Glycans
dc.titleBioanalytical applications of mass spectrometry to identify proteins and glycans from complex biological samples
dc.description.advisorRon Orlando
dc.description.committeeRon Orlando
dc.description.committeeCarl Bergmann
dc.description.committeeJon Amster

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