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Clin Chem. 2016 Jan;62(1):48-69. doi: 10.1373/clinchem.2015.250563.

Recommendations for the Generation, Quantification, Storage, and Handling of Peptides Used for Mass Spectrometry-Based Assays.

Author information

1
University of Washington, Seattle, WA; ahoof@u.washington.edu apaulovi@fhcrc.org.
2
Fred Hutchinson Cancer Research Center, Seattle, WA;
3
Broad Institute, Cambridge, MA;
4
Pacific Northwest National Laboratory, Richland, WA;
5
New England Peptide, Inc., Gardner, MA;
6
Johns Hopkins University, Baltimore, MD;
7
Washington University, St Louis, MO;
8
Vanderbilt University, Nashville, TN;
9
National Cancer Institute, Bethesda, MD;
10
ESAC, Inc., Rockville, MD;
11
Frederick National Laboratory for Cancer Research, Frederick, MD;
12
University of North Carolina School of Medicine, Chapel Hill, NC;
13
Eli Lilly and Company, Indianapolis, IN;
14
Institute of Molecular Systems Biology, ETH Zurich, Zurich, Switzerland;
15
Mayo Clinic College of Medicine, Rochester, MN;
16
NIST, Gaithersburg, MD;
17
Quest Diagnostics, San Juan Capistrano, CA;
18
21st Century Biochemicals, Inc., Marlborough, MA;
19
Laboratory Corporation of America Holdings, Inc., Burlington, NC;
20
Institute for Systems Biology, Seattle, WA;
21
University of Utah and ARUP Laboratories, Salt Lake City, UT;
22
Pfizer, Inc., Andover, MA;
23
Gilead Sciences, Inc., Foster City, CA;
24
Thermo Fisher Scientific, Rockford, IL;
25
Cedars Sinai Medical Center, Los Angeles, CA;
26
Wake Forest School of Medicine, Winston-Salem, NC;
27
Enanta Pharmaceuticals, Watertown, MA;
28
Baylor College of Medicine, Houston, TX.
29
Fred Hutchinson Cancer Research Center, Seattle, WA; ahoof@u.washington.edu apaulovi@fhcrc.org.

Abstract

BACKGROUND:

For many years, basic and clinical researchers have taken advantage of the analytical sensitivity and specificity afforded by mass spectrometry in the measurement of proteins. Clinical laboratories are now beginning to deploy these work flows as well. For assays that use proteolysis to generate peptides for protein quantification and characterization, synthetic stable isotope-labeled internal standard peptides are of central importance. No general recommendations are currently available surrounding the use of peptides in protein mass spectrometric assays.

CONTENT:

The Clinical Proteomic Tumor Analysis Consortium of the National Cancer Institute has collaborated with clinical laboratorians, peptide manufacturers, metrologists, representatives of the pharmaceutical industry, and other professionals to develop a consensus set of recommendations for peptide procurement, characterization, storage, and handling, as well as approaches to the interpretation of the data generated by mass spectrometric protein assays. Additionally, the importance of carefully characterized reference materials-in particular, peptide standards for the improved concordance of amino acid analysis methods across the industry-is highlighted. The alignment of practices around the use of peptides and the transparency of sample preparation protocols should allow for the harmonization of peptide and protein quantification in research and clinical care.

PMID:
26719571
PMCID:
PMC4830481
DOI:
10.1373/clinchem.2015.250563
[Indexed for MEDLINE]
Free PMC Article

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