Internal Standard Triggered-Parallel Reaction Monitoring Mass Spectrometry Enables Multiplexed Quantification of Candidate Biomarkers in Plasma
| Autor: | Jacob J. Kennedy, Jeffrey R. Whiteaker, Richard G. Ivey, Aura Burian, Shrabanti Chowdhury, Chia-Feng Tsai, Tao Liu, ChenWei Lin, Oscar D. Murillo, Rachel A. Lundeen, Lisa A. Jones, Philip R. Gafken, Gary Longton, Karin D. Rodland, Steven J. Skates, John Landua, Pei Wang, Michael T. Lewis, Amanda G. Paulovich |
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| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Analytical Chemistry. 94:9540-9547 |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.1c04382 |
| Popis: | Despite advances in proteomic technologies, clinical translation of plasma biomarkers remains low, partly due to a major bottleneck between the discovery of candidate biomarkers and costly clinical validation studies. Due to a dearth of multiplexable assays, generally only a few candidate biomarkers are tested, and the validation success rate is accordingly low. Previously, mass spectrometry-based approaches have been used to fill this gap but feature poor quantitative performance and were generally limited to hundreds of proteins. Here, we demonstrate the capability of an internal standard triggered-parallel reaction monitoring (IS-PRM) assay to greatly expand the numbers of candidates that can be tested with improved quantitative performance. The assay couples immunodepletion and fractionation with IS-PRM and was developed and implemented in human plasma to quantify 5176 peptides representing 1314 breast cancer biomarker candidates. Characterization of the IS-PRM assay demonstrated the precision (median % CV of 7.7%), linearity (median |
| Databáze: | OpenAIRE |
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