Systematic quantification of peptides/proteins in urine using selected reaction monitoring
| Autor: | Mariette Matondo, Bruno Domon, Ruedi Aebersold, Nathalie Selevsek, Marta Sanchez Carbayo |
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| Přispěvatelé: | Institute of Molecular Systems Biology [Zurich], Eidgenössische Technische Hochschule - Swiss Federal Institute of Technology [Zürich] (ETH Zürich), Spanish National Cancer Research Center (CNIO), Universität Zürich [Zürich] = University of Zurich (UZH), Luxembourg Clinical Proteomics Center [Luxembourg], Luxembourg Institute of Health (LIH), This study was supported by the FP7 Program DECanBio (Coordinator: Jérôme Garin). The Fonds National de la Recherche (FNR) of the Grand Duchy of Luxembourg is acknowledged (PEARL grant)., The authors thank Jérôme Garin, Christophe Masselon, Elodie Duriez, and Martin Soste for their comments and helpful discussions. The authors also thank their clinical collaborators, the hospitals in Salamanca, Oviedo, Barcelona, Guadalajara, and Córdoba for providing the clinical samples analyzed in this study., European Project: 201333,EC:FP7:HEALTH,FP7-HEALTH-2007-A,DECANBIO(2008) |
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Proteomics
MESH: Clusterin / urine Technology Clinical Biochemistry Analytical chemistry MESH: Quality Control MESH: Amino Acid Sequence Urine MESH: Peptides / standards Tandem mass spectrometry Biochemistry 0302 clinical medicine Tandem Mass Spectrometry Multiplex Sample preparation 0303 health sciences Chemistry 030302 biochemistry & molecular biology Bladder cancer Reference Standards Immunohistochemistry MESH: Case-Control Studies MESH: Hyaluronan Receptors / urine MESH: Proteomics / methods MESH: Reproducibility of Results MESH: Urine / chemistry Proteinuria Hyaluronan Receptors 030220 oncology & carcinogenesis Data Interpretation Statistical Selected reaction monitoring MESH: Reference Standards Quality Control MESH: Proteomics / statistics & numerical data Analyte MESH: Tandem Mass Spectrometry / standards Molecular Sequence Data MESH: Peptides / urine MESH: Tandem Mass Spectrometry / statistics & numerical data 03 medical and health sciences MESH: Analysis of Variance MESH: Biomarkers Tumor / urine Biomarkers Tumor Humans [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Amino Acid Sequence MESH: Proteinuria / urine Molecular Biology 030304 developmental biology Reproducibility Analysis of Variance Chromatography MESH: Molecular Sequence Data MESH: Humans Reproducibility of Results MESH: Proteomics / standards MESH: Immunohistochemistry MESH: Urinary Bladder Neoplasms / urine Clusterin Urinary Bladder Neoplasms Case-Control Studies MESH: Peptides / chemistry Peptides Quantitative analysis (chemistry) MESH: Chromatography Liquid MESH: Data Interpretation Statistical Biomarkers Chromatography Liquid |
| Zdroj: | Proteomics Proteomics, Wiley-VCH Verlag, 2011, 11 (6), pp.1135-1147. ⟨10.1002/pmic.201000599⟩ PROTEOMICS; Vol 11 PROTEOMICS |
| ISSN: | 1615-9853 1615-9861 |
| DOI: | 10.1002/pmic.201000599⟩ |
| Popis: | International audience; The evaluation of biomarkers in bodily fluids necessitates the development of robust methods to quantify proteins in a complex background, using large sets of samples. The ability to multiplex numerous analytes in a single assay expedites the process. Liquid chromatography-mass spectrometry (LC-MS) analyses performed in selected reaction monitoring (SRM) in conjunction with stable isotope dilution MS present an effective way to detect and quantify biomarker candidates in bodily fluids. The strategy presented involves an initial qualification of predefined sets of proteins in urine. The technique was applied to detect and quantify peptides in urine samples as surrogates for a few endogenous proteins. Multiplexed assays were developed to analyze proteins associated with bladder cancer; a few exogenous proteins were added as internal standards. The sample preparation and the analytical protocols were optimized to ensure reproducibility, analytical precision, and quantification limits in the low nanogram per milliliter range. Analyses were performed using known amounts of isotopically labeled peptides. Systematic replication of the measurements indicated intra-assay and inter-assay variability, with CVs in the range of 10%. The differences measured for two targeted proteins were correlated with their level of expression in the corresponding tumors using immunohistochemistry. |
| Databáze: | OpenAIRE |
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