title: Advancing the sensitivity of selected reaction monitoring-based targeted quantitative proteomics.
abstract: Selected reaction monitoring (SRM) - also known as multiple reaction monitoring (MRM) - has emerged as a promising high-throughput targeted protein quantification technology for candidate biomarker verification and systems biology applications. A major bottleneck for current SRM technology, however, is insufficient sensitivity for, e.g. detecting low-abundance biomarkers likely present at the low ng/mL to pg/mL range in human blood plasma or serum, or extremely low-abundance signaling proteins in cells or tissues. Herein, we review recent advances in methods and technologies, including front-end immunoaffinity depletion, fractionation, selective enrichment of target proteins/peptides including posttranslational modifications, as well as advances in MS instrumentation which have significantly enhanced the overall sensitivity of SRM assays and enabled the detection of low-abundance proteins at low- to sub-ng/mL level in human blood plasma or serum. General perspectives on the potential of achieving sufficient sensitivity for detection of pg/mL level proteins in plasma are also discussed.
authors.affiliation: Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors.forename: Tujin
authors.initials: T
authors.lastname: Shi
authors.forename: Dian
authors.initials: D
authors.lastname: Su
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.forename: Keqi
authors.initials: K
authors.lastname: Tang
authors.forename: David G
authors.initials: DG
authors.lastname: Camp
authors.forename: Wei-Jun
authors.initials: WJ
authors.lastname: Qian
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
mesh_terms: D015415:Biomarkers / Q000032:analysis*; D001798:Blood Proteins / Q000032:analysis*; D005591:Chemical Fractionation; D057166:High-Throughput Screening Assays; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D015415:Biomarkers; D001798:Blood Proteins