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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Giménez, Estela Balmaña, Meritxell Figueras, Joan Fort, Esther de Bolós, Carme Sanz-Nebot, Victòria Peracaula, Rosa Rizzi, Andreas |
| Description | Author Affiliation: Giménez E ( Department of Analytical Chemistry, University of Barcelona, Diagonal 647, E-08028 Barcelona, Spain. Electronic address: estelagimenez@ub.edu.); Balmaña M ( Biochemistry and Molecular Biology Unit, Department of Biology, University of Girona, Campus Montilivi s/n, 17071 Girona, Spain.); Figueras J ( Department of Surgery, Dr. Josep Trueta University Hospital, IdlBGi, 17007 Girona, Spain.); Fort E ( Digestive Unit, Dr. Josep Trueta University Hospital, 17007 Girona, Spain.); de Bolós C ( Gastroesophagic Cancer Research Group, Research Programme in Cancer, Hospital del Mar Medical Research Institute (IMIM), Dr. Aiguader, 88, 08003 Barcelona, Spain.); Sanz-Nebot V ( Department of Analytical Chemistry, University of Barcelona, Diagonal 647, E-08028 Barcelona, Spain.); Peracaula R ( Biochemistry and Molecular Biology Unit, Department of Biology, University of Girona, Campus Montilivi s/n, 17071 Girona, Spain.); Rizzi A ( Institute of Analytical Chemistry, University of Vienna, Währinger Straße 38, A-1090 Vienna, Austria.) |
| Abstract | In this work we demonstrate the potential of glycan reductive isotope labeling (GRIL) using [(12)C]- and [(13)C]-coded aniline and zwitterionic hydrophilic interaction capillary liquid chromatography electrospray mass spectrometry (µZIC-HILIC-ESI-MS) for relative quantitation of glycosylation variants in selected glycoproteins present in samples from cancer patients. Human 1-acid-glycoprotein (hAGP) is an acute phase serum glycoprotein whose glycosylation has been described to be altered in cancer and chronic inflammation. However, it is not clear yet whether some particular glycans in hAGP can be used as biomarker for differentiating between these two pathologies. In this work, hAGP was isolated by immunoaffinity chromatography (IAC) from serum samples of healthy individuals and from those suffering chronic pancreatitis and different stages of pancreatic cancer, respectively. After de-N-glycosylation, relative quantitation of the hAGP glycans was carried out using stable isotope labeling and µZIC-HILIC-ESI-MS analysis. First, protein denaturing conditions prior to PNGase F digestion were optimized to achieve quantitative digestion yields, and the reproducibility of the established methodology was evaluated with standard hAGP. Then, the proposed method was applied to the analysis of the clinical samples (control vs. pathological). Pancreatic cancer samples clearly showed an increase in the abundance of fucosylated glycans as the stage of the disease increases and this was unlike to samples from chronic pancreatitis. The results gained here indicate the mentioned glycan in hAGP as a candidate structure worth to be corroborated by an extended study including more clinical cases; especially those with chronic pancreatitis and initial stages of pancreatic cancer. Importantly, the results demonstrate that the presented methodology combining an enrichment of a target protein by IAC with isotope coded relative quantitation of N-glycans can be successfully used for targeted glycomics studies. The methodology is assumed being suitable as well for other such studies aimed at finding novel cancer associated glycoprotein biomarkers. |
| File Format | HTM / HTML |
| ISSN | 00032670 |
| Volume Number | 866 |
| e-ISSN | 18734324 |
| Journal | Analytica Chimica Acta |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2015-03-25 |
| Publisher Place | Netherlands |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Analytical Discipline Chemistry Orosomucoid Chemistry Polysaccharides Analysis Spectrometry, Mass, Electrospray Ionization Methods Biological Markers Blood Chromatography, Affinity Chromatography, High Pressure Liquid Glycosylation Humans Hydrophobic And Hydrophilic Interactions Isotope Labeling Nitrogen Isolation & Purification Metabolism Pancreatic Neoplasms Pathology Instrumentation Journal Article Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Spectroscopy Environmental Chemistry Analytical Chemistry Biochemistry |
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