Simultaneous Proteomic Discovery and Targeted Monitoring using Liquid Chromatography, Ion Mobility Spectrometry, and Mass Spectrometry

Current proteomic approaches include both broad discovery measurements and quantitative targeted analyses. In many cases, discovery measurements are initially used to identify potentially important proteins (e.g. candidate biomarkers) and then targeted studies are employed to quantify a limited numb...

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Vydáno v:Molecular & cellular proteomics Ročník 15; číslo 12; s. 3694 - 3705
Hlavní autoři: Burnum-Johnson, Kristin E, Nie, Song, Casey, Cameron P, Monroe, Matthew E, Orton, Daniel J, Ibrahim, Yehia M, Gritsenko, Marina A, Clauss, Therese R W, Shukla, Anil K, Moore, Ronald J, Purvine, Samuel O, Shi, Tujin, Qian, Weijun, Liu, Tao, Baker, Erin S, Smith, Richard D
Médium: Journal Article
Jazyk:angličtina
Vydáno: United States 01.12.2016
Témata:
Animals
Breast Neoplasms - metabolism
Chromatography, Liquid - methods
Female
Humans
Mass Spectrometry - methods
Mice
Neoplasm Transplantation
Peptides - analysis
Proteome - isolation & purification
Proteomics - methods
ISSN:1535-9484
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Shrnutí:Current proteomic approaches include both broad discovery measurements and quantitative targeted analyses. In many cases, discovery measurements are initially used to identify potentially important proteins (e.g. candidate biomarkers) and then targeted studies are employed to quantify a limited number of selected proteins. Both approaches, however, suffer from limitations. Discovery measurements aim to sample the whole proteome but have lower sensitivity, accuracy, and quantitation precision than targeted approaches, whereas targeted measurements are significantly more sensitive but only sample a limited portion of the proteome. Herein, we describe a new approach that performs both discovery and targeted monitoring (DTM) in a single analysis by combining liquid chromatography, ion mobility spectrometry and mass spectrometry (LC-IMS-MS). In DTM, heavy labeled target peptides are spiked into tryptic digests and both the labeled and unlabeled peptides are detected using LC-IMS-MS instrumentation. Compared with the broad LC-MS discovery measurements, DTM yields greater peptide/protein coverage and detects lower abundance species. DTM also achieved detection limits similar to selected reaction monitoring (SRM) indicating its potential for combined high quality discovery and targeted analyses, which is a significant step toward the convergence of discovery and targeted approaches.
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ISSN:1535-9484
DOI:10.1074/mcp.M116.061143