| Zugriffsnummer | 32072 |
| Dokumenttyp | Zeitschriftenartikel |
| Sprache | Englisch |
| Titel | Determination of cystatin C in human serum by isotope dilution mass spectrometry using mass overlapping peptides. |
| Autor(in); Institution |
González Antuna, Ana; University of Uviedo, Department of Physical and Analytical Chemistry, Oviedo, SPAIN
Rodríguez-González, Pablo; University of Uviedo, Department of Physical and Analytical Chemistry, Oviedo, SPAIN
Ohlendorf, Rüdiger; 3.1, Metrologie in der Chemie, PTB-Braunschweig
Henrion, André; 3.1, Metrologie in der Chemie, PTB-Braunschweig
Delatour, Vincent; Laboratoire Ntional de Métrologie et d’Essais (LNE), Paris, FRANCE
García Alonso, José Ignacio; University of Uviedo, Department of Physical and Analytical Chemistry, Oviedo, SPAIN
|
| Quelle/Jahr | Journal of Proteomics: 112 (2015), 141 - 155 |
| ISSN | 1874-3919 (PRINT) ; 1876-7737 (ONLINE) |
| DOI | |
| Verlag | Amsterdam [u.a.]: Elsevier |
| Freie Schlagworte | cystatin C ; clinical biomarker ; isotope dilution mass spectrometry ; mass overlapping peptides ; selected reaction monitoring ; absolute protein quantification |
| Zusammenfassung | We propose a peptide-based Isotope Dilution Mass Spectrometry appraoach for the determination of Cystatin C in human serum samples, a clinical marker for renal status, for which backup by a mass spectrometry based primary method for reliable determination has been missing so far. In contrast to common protocols, the isotope labeled version of the proteotypic signature peptide is designed such as keeping the isotopic difference as little as possible with respect to the peptide released from the protein (Mass Overlapping Peptides, MOPs). For Cystatin C we employ MOPs labeled in two 13C atoms which are added to the serum samples just before proteolysis. After two steps of chromatographic purification (reverse phase and ion exchange) the sample is measured by LC-MS/MS in a triple quadrupole instrument. Molar fractions of labeled and natural abundance peptides are directly obtained from the experimental mass spectra of the in-cell fragment ions obtained by selected reaction monitoring (SRM). Resolution of the first quadrupole is reduced to the extent that, the whole parent ion cluster is transmitted to the collision cell for monitoring accurate isotopic distribution of the molecular fragments. |
Zitierung
González Antuna, A., Rodríguez-González, P., Ohlendorf, R., Henrion, A., Delatour, V., & García Alonso, J. I. (2015). Determination of cystatin C in human serum by isotope dilution mass spectrometry using mass overlapping peptides. Journal of Proteomics, 112, 141–155. https://doi.org/10.1016/j.jprot.2014.09.005