Design, Implementation and Multisite Evaluation of a System Suitability Protocol for the Quantitative Assessment of Instrument Performance in Liquid Chromatography-Multiple Reaction Monitoring-MS (LC-MRM-MS)

被引：81

作者：

Abbatiello, Susan E. ^{[1
]}

Mani, D. R. ^{[1
]}

Schilling, Birgit ^{[2
]}

MacLean, Brendan ^{[3
]}

Zimmerman, Lisa J. ^{[4
]}

Feng, Xingdong ^{[5
]}

Cusack, Michael P. ^{[2
]}

Sedransk, Nell ^{[5
]}

Hall, Steven C. ^{[6
]}

Addona, Terri ^{[1
]}

Allen, Simon ^{[6
]}

Dodder, Nathan G. ^{[7
]}

Ghosh, Mousumi ^{[8
]}

Held, Jason M. ^{[2
]}

Hedrick, Victoria ^{[9
]}

Inerowicz, H. Dorota ^{[9
]}

Jackson, Angela ^{[10
]}

Keshishian, Hasmik ^{[1
]}

Kim, Jong Won ^{[11
]}

Lyssand, John S. ^{[12
,13
]}

Riley, C. Paige ^{[9
]}

Rudnick, Paul ^{[7
]}

Sadowski, Pawel ^{[12
,13
]}

Shaddox, Kent ^{[4
]}

Smith, Derek ^{[10
]}

Tomazela, Daniela ^{[3
]}

Wahlander, Asa ^{[12
,13
]}

Waldemarson, Sofia ^{[12
,13
]}

Whitwell, Corbin A. ^{[4
]}

You, Jinsam ^{[11
]}

Zhang, Shucha ^{[14
]}

Kinsinger, Christopher R. ^{[15
]}

Mesri, Mehdi ^{[15
]}

Rodriguez, Henry ^{[15
]}

Borchers, Christoph H. ^{[10
]}

Buck, Charles ^{[9
]}

Fisher, Susan J. ^{[6
]}

Gibson, Bradford W. ^{[2
]}

Liebler, Daniel ^{[4
]}

MacCoss, Michael ^{[3
]}

Neubert, Thomas A. ^{[12
,13
]}

Paulovich, Amanda ^{[14
]}

Regnier, Fred ^{[9
]}

Skates, Steven J. ^{[16
]}

Tempst, Paul ^{[8
]}

Wang, Mu ^{[11
]}

Carr, Steven A. ^{[1
]}

机构：

[1] Broad Inst MIT & Harvard, Cambridge, MA 02142 USA

[2] Buck Inst Res Aging, Novato, CA 94945 USA

[3] Univ Washington, Sch Med, Dept Genome Sci, Seattle, WA 98195 USA

[4] Vanderbilt Univ, Nashville, TN 37232 USA

[5] Natl Inst Stat Sci, Res Triangle Pk, NC 27709 USA

[6] Univ Calif San Francisco, Dept Obstet Gynecol & Reprod Sci, San Francisco, CA 94143 USA

[7] NIST, Gaithersburg, MD 20899 USA

[8] Mem Sloan Kettering Canc Ctr, New York, NY 10065 USA

[9] Purdue Univ, W Lafayette, IN 47907 USA

[10] Univ Victoria, Genome BC Prote Ctr, Victoria, BC V8Z 7X8, Canada

[11] Indiana Univ Sch Med, Dept Biochem & Mol Biol, Indianapolis, IN 46202 USA

[12] NYU, Sch Med, Kimmel Ctr Biol & Med, Skirball Inst, New York, NY 10016 USA

[13] NYU, Sch Med, Dept Pharmacol, New York, NY 10016 USA

[14] Fred Hutchinson Canc Res Ctr, Seattle, WA 98109 USA

[15] NCI, NIH, Bethesda, MD 20892 USA

[16] Massachusetts Gen Hosp, Biostat Ctr, Boston, MA 02114 USA

来源：

MOLECULAR & CELLULAR PROTEOMICS | 2013年 / 12卷 / 09期

关键词：

PEPTIDE IMMUNOAFFINITY ENRICHMENT; MASS-SPECTROMETRIC QUANTIFICATION; ABSOLUTE QUANTIFICATION; TARGETED PROTEOMICS; QUALITY-CONTROL; PROTEINS; PLASMA; ASSAYS;

D O I：

10.1074/mcp.M112.027078

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Multiple reaction monitoring (MRM) mass spectrometry coupled with stable isotope dilution (SID) and liquid chromatography (LC) is increasingly used in biological and clinical studies for precise and reproducible quantification of peptides and proteins in complex sample matrices. Robust LC-SID-MRM-MS-based assays that can be replicated across laboratories and ultimately in clinical laboratory settings require standardized protocols to demonstrate that the analysis platforms are performing adequately. We developed a system suitability protocol (SSP), which employs a predigested mixture of six proteins, to facilitate performance evaluation of LC-SID-MRM-MS instrument platforms, configured with nanoflow-LC systems interfaced to triple quadrupole mass spectrometers. The SSP was designed for use with low multiplex analyses as well as high multiplex approaches when software-driven scheduling of data acquisition is required. Performance was assessed by monitoring of a range of chromatographic and mass spectrometric metrics including peak width, chromatographic resolution, peak capacity, and the variability in peak area and analyte retention time (RT) stability. The SSP, which was evaluated in 11 laboratories on a total of 15 different instruments, enabled early diagnoses of LC and MS anomalies that indicated suboptimal LC-MRM-MS performance. The observed range in variation of each of the metrics scrutinized serves to define the criteria for optimized LCSID- MRM-MS platforms for routine use, with pass/ fail criteria for system suitability performance measures defined as peak area coefficient of variation < 0.15, peak width coefficient of variation < 0.15, standard deviation of RT < 0.15 min (9 s), and the RT drift < 0.5min (30 s). The deleterious effect of a marginally performing LC-SID-MRM-MS system on the limit of quantification (LOQ) in targeted quantitative assays illustrates the use and need for a SSP to establish robust and reliable system performance. Use of a SSP helps to ensure that analyte quantification measurements can be replicated with good precision within and across multiple laboratories and should facilitate more widespread use of MRM-MS technology by the basic biomedical and clinical laboratory research communities.

引用

页码：2623 / 2639

页数：17

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