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ABSTRACT Title of dissertation: CAPILLARY ISOELECTRIC FOCUSING-BASED MULTIDIMENSIONAL PEPTIDE/PROTEIN SEPARATIONS FOR PROTEOMICS ANALYSIS
| Content Provider | CiteSeerX |
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| Abstract | With the completion of the human genome project, the proteomics has become the focus of research interest for better understanding the complex biological processes. The mass spectrometry (MS) detection of vast number and broad dynamic range of the proteins requires that sample fractionation and separation be performed prior to MS analysis. Realizing the limitations of gel-based proteomic techniques, capillary-based and gel free separation technologies are presented as attractive alternatives holding the promises of high separation efficiency and resolution, broad dynamic range, easy system automation as well as high throughput. Coupled with laser free microdissection technology, the combination of CIEF with nano-RPLC in an automated and integrated platform was employed for comprehensive and sensitive proteome studies of limited protein quantities obtained from tissue samples. The peptides were first separated and concentrated by CIEF and were sequentially fractionated. All the CIEF fractions were further resolved by nano-RPLC, followed by tandem MS analysis. A total of 6,866 fully tryptic peptides were detected, leading to the identification of 1,820 distinct proteins. |
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