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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Abbatiello, Susan E. Schilling, Birgit Mani, D. R. Zimmerman, Lisa J. Hall, Steven C. MacLean, Brendan Albertolle, Matthew Allen, Simon Burgess, Michael Cusack, Michael P. Gosh, Mousumi Hedrick, Victoria Held, Jason M. Inerowicz, H. Dorota Jackson, Angela Keshishian, Hasmik Kinsinger, Christopher R. Lyssand, John Makowski, Lee Mesri, Mehdi Rodriguez, Henry Rudnick, Paul Sadowski, Pawel Sedransk, Nell Shaddox, Kent Skates, Stephen J. Kuhn, Eric Smith, Derek Whiteaker, Jeffery R. Whitwell, Corbin Zhang, Shucha Borchers, Christoph H. Fisher, Susan J. Gibson, Bradford W. Liebler, Daniel C. MacCoss, Michael J. Neubert, Thomas A. Paulovich, Amanda G. Regnier, Fred E. Tempst, Paul Carr, Steven A. |
| Description | Author Affiliation: Abbatiello SE ( From the Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142); Schilling B ( Buck Institute for Research on Aging, Novato, California 94945); Mani DR ( From the Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142); Zimmerman LJ ( Department of Biochemistry, Vanderbilt University School of Medicine, and the Jim Ayers Institute for Precancer Detection and Diagnosis, Vanderbilt-Ingram Cancer Center, Nashville, Tennessee 37232); Hall SC ( Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, San Francisco, California 94143); MacLean B ( Department of Genome Sciences, University of Washington School of Medicine, Seattle, Washington 98195); Albertolle M ( Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, San Francisco, California 94143); Allen S ( Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, San Francisco, California 94143); Burgess M ( From the Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142); Cusack MP ( Buck Institute for Research on Aging, Novato, California 94945); Gosh M ( Memorial Sloan-Kettering Cancer Center, New York, New York 10065); Hedrick V ( Purdue University, West Lafayette, Indiana 47907); Held JM ( Buck Institute for Research on Aging, Novato, California 94945); Inerowicz HD ( Purdue University, West Lafayette, Indiana 47907); Jackson A ( University of Victoria-Genome BC Proteomics Centre, Victoria, British Columbia V8Z 7X8 CAN); Keshishian H ( From the Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142); Kinsinger CR ( National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892); Lyssand J ( Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, New York 10016); Makowski L ( Argonne National Laboratory (currently at Northeastern University, Boston Massachusetts 02115); Mesri M ( National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892); Rodriguez H ( National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892); Rudnick P ( National Institute of Standards and Technology, Gaithersburg, Maryland 20899); Sadowski P ( Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, New York 10016); Sedransk N ( National Institute of Statistical Sciences, Research Triangle Park, North Carolina 27709); Shaddox K ( Department of Biochemistry, Vanderbilt University School of Medicine, and the Jim Ayers Institute for Precancer Detection and Diagnosis, Vanderbilt-Ingram Cancer Center, Nashville, Tennessee 37232); Skates SJ ( Biostatistics Center, Massachusetts General Hospital, Boston, Massachusetts 02114); Kuhn E ( From the Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142); Smith D ( University of Victoria-Genome BC Proteomics Centre, Victoria, British Columbia V8Z 7X8 CAN); Whiteaker JR ( Fred Hutchinson Cancer Research Center, Seattle, Washington 98109.); Whitwell C ( Department of Biochemistry, Vanderbilt University School of Medicine, and the Jim Ayers Institute for Precancer Detection and Diagnosis, Vanderbilt-Ingram Cancer Center, Nashville, Tennessee 37232); Zhang S ( Fred Hutchinson Cancer Research Center, Seattle, Washington 98109.); Borchers CH ( University of Victoria-Genome BC Proteomics Centre, Victoria, British Columbia V8Z 7X8 CAN); Fisher SJ ( Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California, San Francisco, San Francisco, California 94143); Gibson BW ( Buck Institute for Research on Aging, Novato, California 94945); Liebler DC ( Department of Biochemistry, Vanderbilt University School of Medicine, and the Jim Ayers Institute for Precancer Detection and Diagnosis, Vanderbilt-Ingram Cancer Center, Nashville, Tennessee 37232); MacCoss MJ ( Department of Genome Sciences, University of Washington School of Medicine, Seattle, Washington 98195); Neubert TA ( Kimmel Center for Biology and Medicine at the Skirball Institute, New York University School of Medicine, New York, New York 10016); Paulovich AG ( Fred Hutchinson Cancer Research Center, Seattle, Washington 98109.); Regnier FE ( Purdue University, West Lafayette, Indiana 47907); Tempst P ( Memorial Sloan-Kettering Cancer Center, New York, New York 10065); Carr SA ( From the Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142) |
| Abstract | There is an increasing need in biology and clinical medicine to robustly and reliably measure tens to hundreds of peptides and proteins in clinical and biological samples with high sensitivity, specificity, reproducibility, and repeatability. Previously, we demonstrated that LC-MRM-MS with isotope dilution has suitable performance for quantitative measurements of small numbers of relatively abundant proteins in human plasma and that the resulting assays can be transferred across laboratories while maintaining high reproducibility and quantitative precision. Here, we significantly extend that earlier work, demonstrating that 11 laboratories using 14 LC-MS systems can develop, determine analytical figures of merit, and apply highly multiplexed MRM-MS assays targeting 125 peptides derived from 27 cancer-relevant proteins and seven control proteins to precisely and reproducibly measure the analytes in human plasma. To ensure consistent generation of high quality data, we incorporated a system suitability protocol (SSP) into our experimental design. The SSP enabled real-time monitoring of LC-MRM-MS performance during assay development and implementation, facilitating early detection and correction of chromatographic and instrumental problems. Low to subnanogram/ml sensitivity for proteins in plasma was achieved by one-step immunoaffinity depletion of 14 abundant plasma proteins prior to analysis. Median intra- and interlaboratory reproducibility was <20%, sufficient for most biological studies and candidate protein biomarker verification. Digestion recovery of peptides was assessed and quantitative accuracy improved using heavy-isotope-labeled versions of the proteins as internal standards. Using the highly multiplexed assay, participating laboratories were able to precisely and reproducibly determine the levels of a series of analytes in blinded samples used to simulate an interlaboratory clinical study of patient samples. Our study further establishes that LC-MRM-MS using stable isotope dilution, with appropriate attention to analytical validation and appropriate quality control measures, enables sensitive, specific, reproducible, and quantitative measurements of proteins and peptides in complex biological matrices such as plasma. |
| File Format | HTM / HTML |
| ISSN | 15359476 |
| e-ISSN | 15359484 |
| DOI | 10.1074/mcp.M114.047050 |
| Journal | Molecular & Cellular Proteomics |
| Issue Number | 9 |
| Volume Number | 14 |
| Language | English |
| Publisher | American Society for Biochemistry and Molecular Biology |
| Publisher Date | 2015-09-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Discipline Proteomics Neoplasm Proteins Blood Neoplasms Metabolism Peptides Proteomics Chromatography, Liquid Isotope Labeling Mass Spectrometry Chemistry Isolation & Purification Reproducibility Of Results Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Analytical Chemistry Molecular Biology Biochemistry |
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