Deep-Dive Targeted Quantification for Ultrasensitive Analysis of Proteins in Nondepleted Human Blood Plasma/Serum and Tissues

Journal Article · Fri Aug 11 00:00:00 EDT 2017 · Analytical Chemistry

DOI:https://doi.org/10.1021/acs.analchem.7b01878· OSTI ID:1406735

Nie, Song ^[1]; Shi, Tujin ^[1]; Fillmore, Thomas L. ^[1]; Schepmoes, Athena A. ^[1]; Brewer, Heather ^[1]; Gao, Yuqian ^[1]; Song, Ehwang ^[1]; Wang, Hui ^[1]; Rodland, Karin D. ^[1]; Qian, Wei-Jun ^[1]; Smith, Richard D.  ^[1]; Liu, Tao  ^[1]

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1. Biological Sciences Division and Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, United States

Mass spectrometry-based targeted proteomics (e.g., selected reaction monitoring, SRM) is emerging as an attractive alternative to immunoassays for protein quantification. Recently we have made significant progress in SRM sensitivity for enabling quantification of low ng/mL to sub-ng/mL level proteins in nondepleted human blood plasma/serum without affinity enrichment. However, precise quantification of extremely low abundant but biologically important proteins (e.g., ≤100 pg/mL in blood plasma/serum) using targeted proteomics approaches still remains challenging. To address this need, we have developed an antibody-independent Deep-Dive SRM (DD-SRM) approach that capitalizes on multidimensional high-resolution reversed-phase liquid chromatography (LC) separation for target peptide enrichment combined with precise selection of target peptide fractions of interest, significantly improving SRM sensitivity by ~5 orders of magnitude when compared to conventional LC-SRM. Application of DD-SRM to human serum and tissue has been demonstrated to enable precise quantification of endogenous proteins at ~10 pg/mL level in nondepleted serum and at <10 copies per cell level in tissue. Thus, DD-SRM holds great promise for precisely measuring extremely low abundance proteins or protein modifications, especially when high-quality antibody is not available.

• Research Organization: Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
• Sponsoring Organization: USDOE
• DOE Contract Number: AC05-76RL01830
• OSTI ID: 1406735
• Report Number(s): PNNL-SA-126297; 48666; 453040220
• Journal Information: Analytical Chemistry, Vol. 89, Issue 17; ISSN 0003-2700
• Publisher: American Chemical Society (ACS)
• Country of Publication: United States
• Language: English

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Environmental Molecular Sciences Laboratory