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Assessment of Universal Approaches to Proteome Prefractionation

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dc.contributor.author Liu, Fang
dc.date.accessioned 2011-06-15T11:24:53Z
dc.date.available 2011-06-15T11:24:53Z
dc.date.issued 2011-06-15
dc.identifier.uri http://hdl.handle.net/10222/13791
dc.description.abstract Protein prefractionation is a popular and effective strategy for improved MS analysis of complex proteome mixtures. A challenge of prefractionation is the even partitioning with high recovery of all components of the mixture, particularly hydrophobic proteins. This thesis assesses various proteome prefractionation platforms, with a goal of comprehensive proteome analysis. A more reliable dataset of 1136 S. cerevisiae transmembrane proteins was computationally generated, and used to assess two gel-based platforms (GeLC/MS and GELFrEE/MS). These platforms were determined to be comparable for proteome analysis. The requirement for high-throughput, automated fractionation demands a gel-free separation workflow. Here, a LC-based workflow was optimized, relying on SDS-assisted yeast extraction, organic solvent protein precipitation, and reversed phase separation in a formic acid/isopropanol solvent system. Though this workflow afforded improvements over conventional LC strategies to proteome fractionation, the gel-based platforms were demonstrated to be superior, in terms of their unbiased separation of hydrophobic vs hydrophilic proteins. en_US
dc.language.iso en en_US
dc.subject Proteome, Prefractionation, GeLC, GELFrEE, RPLC, Mass Spectrometry, Protein Identification en_US
dc.title Assessment of Universal Approaches to Proteome Prefractionation en_US
dc.date.defence 2011-05-13
dc.contributor.department Department of Chemistry en_US
dc.contributor.degree Master of Science en_US
dc.contributor.external-examiner Dr. Peng Zhang en_US
dc.contributor.graduate-coordinator Dr. Mark Stradiotto en_US
dc.contributor.thesis-reader Dr. Robert D. Guy, Dr. Peter D. Wentzell en_US
dc.contributor.thesis-supervisor Dr. Alan A. Doucette en_US
dc.contributor.ethics-approval Not Applicable en_US
dc.contributor.manuscripts Not Applicable en_US
dc.contributor.copyright-release Not Applicable en_US


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