Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.14/184893
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- Title
- The Development of an integrated platform to identify breast cancer glycoproteome changes in human serum
- Related
- Journal of chromatography A, Vol. 1217, Issue 19, (2010), p.3307-3315
- DOI
- 10.1016/j.chroma.2009.09.029
- Publisher
- Elsevier BV
- Date
- 2010
- FoR/RFCD Code(s)
-
030100 Analytical Chemistry
- Author/Creator
- Zeng, Zhi
- Author/Creator
- Hincapie, Marina
- Author/Creator
- Haab, Brian B
- Author/Creator
- Hanash, Samir
- Author/Creator
- Pitteri, Sharon J
- Author/Creator
- Kluck, Steven
- Author/Creator
- Hogan, Jason M
- Author/Creator
- Kennedy, Jacob
- Author/Creator
- Hancock, William S
- Description
- Protein glycosylation represents one of the major post-translational modifications and can have significant effects on protein function. Moreover, changes in the carbohydrate structure are increasingly being recognized as an important modification associated with cancer etiology. In this report, we describe the development of a proteomics approach to identify breastcancer related changes in either concentration and/or the carbohydrate structures of glycoprotein(s) present in blood samples. Diseased and healthy serum samples were processed by an optimized sample preparation protocol using multiple lectin affinity chromatography (M-LAC) that partitions serum proteins based on glycan characteristics. Subsequently, three separate procedures, 1D SDS-PAGE, isoelectric focusing and an antibody microarray, were applied to identify potential candidate markers for future study. The combination of these three platforms is illustrated in this report with the analysis of control and cancer glycoproteomic fractions. Firstly, a molecular weight based separation of glycoproteins by 1D SDS-PAGE was performed, followed by protein, glycoprotein staining, lectin blotting and LC–MS analysis. To refine or confirm the list of interesting glycoproteins, isoelectric focusing (targeting sialic acid changes) and an antibody microarray (used to detect neutral glycan shifts) were selected as the orthogonal methods. As a result, several glycoproteins including alpha-1B-glycoprotein, complement C3, alpha-1-antitrypsin and transferrin were identified as potential candidates for further study.
- Description
- 9 page(s)
- Subject Keyword
- 030100 Analytical Chemistry
- Subject Keyword
- High-performance multi-lectin affinity chromatography
- Subject Keyword
- Lectin blotting
- Subject Keyword
- Isoelectric focusing
- Subject Keyword
- Lectin–antibody microarray
- Resource Type
- journal article
- Organisation
- Macquarie University. Dept. of Chemistry and Biomolecular Sciences
- Identifier
- http://hdl.handle.net/1959.14/184893
- Identifier
- ISSN:0021-9673
- Identifier
- mq-rm-2011000520
- Language
- eng
- Reviewed
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