Platform for Establishing Interlaboratory Reproducibility of Selected Reaction Monitoring-Based Mass Spectrometry Peptide Assays
| Autor: | L. Liotta, Eleftherios P. Diamandis, Amol Prakash, M. M. Ross, Emanuel F. Petricoin, Bryan Krastins, Michael Athanas, H. Zhang, Taha Rezai, David A. Sarracino, Y. Tian, D. W. Chan, P. Russo, Andrei P. Drabovich, Vathany Kulasingam, Mary F. Lopez, I. Batruch, C. Smith |
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| Rok vydání: | 2010 |
| Předmět: |
chemistry.chemical_classification
Reproducibility Chromatography Interlaboratory reproducibility Clinical Laboratory Techniques Coefficient of variation Selected reaction monitoring Analytical chemistry Proteins Reproducibility of Results Peptide General Chemistry Reference Standards Mass spectrometry Proteomics Biochemistry Mass Spectrometry Matrix (chemical analysis) chemistry North America Humans Amino Acid Sequence Peptides |
| Zdroj: | Journal of Proteome Research. 9:6678-6688 |
| ISSN: | 1535-3907 1535-3893 |
| Popis: | Mass spectrometry (MS) is an attractive alternative to quantification of proteins by immunoassays, particularly for protein biomarkers of clinical relevance. Reliable quantification requires that the MS-based assays are robust, selective, and reproducible. Thus, the development of standardized protocols is essential to introduce MS into clinical research laboratories. The aim of this study was to establish a complete workflow for assessing the transferability and reproducibility of selected reaction monitoring (SRM) assays between clinical research laboratories. Four independent laboratories in North America, using identical triple-quadrupole mass spectrometers (Quantum Ultra, Thermo), were provided with standard protocols and instrumentation settings to analyze unknown samples and internal standards in a digested plasma matrix to quantify 51 peptides from 39 human proteins using a multiplexed SRM assay. The interlaboratory coefficient of variation (CV) was less than 10% for 25 of 39 peptides quantified (12 peptides were not quantified based upon hydrophobicity) and exhibited CVs less than 20% for the remaining peptides. In this report, we demonstrate that previously developed research platforms for SRM assays can be improved and optimized for deployment in clinical research environments. |
| Databáze: | OpenAIRE |
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