Demonstrating the feasibility of large-scale development of standardized assays to quantify human proteins.

Nat Methods
Authors
Keywords
Abstract

Multiple reaction monitoring (MRM) mass spectrometry has been successfully applied to monitor targeted proteins in biological specimens, raising the possibility that assays could be configured to measure all human proteins. We report the results of a pilot study designed to test the feasibility of a large-scale, international effort for MRM assay generation. We have configured, validated across three laboratories and made publicly available as a resource to the community 645 novel MRM assays representing 319 proteins expressed in human breast cancer. Assays were multiplexed in groups of >150 peptides and deployed to quantify endogenous analytes in a panel of breast cancer-related cell lines. The median assay precision was 5.4%, with high interlaboratory correlation (R(2) > 0.96). Peptide measurements in breast cancer cell lines were able to discriminate among molecular subtypes and identify genome-driven changes in the cancer proteome. These results establish the feasibility of a large-scale effort to develop an MRM assay resource.

Year of Publication
2014
Journal
Nat Methods
Volume
11
Issue
2
Pages
149-55
Date Published
2014 Feb
ISSN
1548-7105
URL
DOI
10.1038/nmeth.2763
PubMed ID
24317253
PubMed Central ID
PMC3922286
Links
Grant list
U24 CA160034 / CA / NCI NIH HHS / United States
R01 GM082802 / GM / NIGMS NIH HHS / United States
P50 CA138293 / CA / NCI NIH HHS / United States
RC2CA148286 / CA / NCI NIH HHS / United States
RC2 CA148286 / CA / NCI NIH HHS / United States
U24CA160034 / CA / NCI NIH HHS / United States
P50CA138293 / CA / NCI NIH HHS / United States