ArticleMolecular & cellular proteomics : MCP2014
Simplified and efficient quantification of low-abundance proteins at very high multiplex via targeted mass spectrometry.
Article in Molecular & cellular proteomics : MCP, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 32 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
32 citing papers in PubMed.
- Article
- Instrumentation at the Leading Edge of Proteomics.Analytical chemistry · 2024Review
- Quantitative Cell Proteomic Atlas: Pathway-Scale Targeted Mass Spectrometry for High-Resolution Functional Profiling of Cell Signaling.Journal of proteome research · 2022Article
- Review
- Internal Standard Triggered-Parallel Reaction Monitoring Mass Spectrometry Enables Multiplexed Quantification of Candidate Biomarkers in Plasma.Analytical chemistry · 2022Article
- A highly multiplexed quantitative phosphosite assay for biology and preclinical studies.Molecular systems biology · 2021Article
- Targeted mass spectrometry-based assays enable multiplex quantification of receptor tyrosine kinase, MAP Kinase, and AKT signaling.Cell reports methods · 2021Article
- Use of Hybrid Data-Dependent and -Independent Acquisition Spectral Libraries Empowers Dual-Proteome Profiling.Journal of proteome research · 2021Article
- An Introduction to Advanced Targeted Acquisition Methods.Molecular & cellular proteomics : MCP · 2021Review
- An MRM-Based Multiplexed Quantification Assay for Human Adipokines and Apolipoproteins.Molecules (Basel, Switzerland) · 2020Article
- Rapidly Assessing the Quality of Targeted Proteomics Experiments through Monitoring Stable-Isotope Labeled Standards.Journal of proteome research · 2019Article
- Quantification of Cardiovascular Disease Biomarkers in Human Platelets by Targeted Mass Spectrometry.Proteomes · 2017Article
- High Sensitivity Quantitative Proteomics Using Automated Multidimensional Nano-flow Chromatography and Accumulated Ion Monitoring on Quadrupole-Orbitrap-Linear Ion Trap Mass Spectrometer.Molecular & cellular proteomics : MCP · 2017Article
- Revisiting biomarker discovery by plasma proteomics.Molecular systems biology · 2017Review
- Quantitative, multiplexed workflow for deep analysis of human blood plasma and biomarker discovery by mass spectrometry.Nature protocols · 2017Article
- An Optimized Chromatographic Strategy for Multiplexing In Parallel Reaction Monitoring Mass Spectrometry: Insights from Quantitation of Activated Kinases.Molecular & cellular proteomics : MCP · 2017Article
- Psychiatric disorders biochemical pathways unraveled by human brain proteomics.European archives of psychiatry and clinical neuroscience · 2017Review
- A Strategy to Combine Sample Multiplexing with Targeted Proteomics Assays for High-Throughput Protein Signature Characterization.Molecular cell · 2017Article
- Transitioning from Targeted to Comprehensive Mass Spectrometry Using Genetic Algorithms.Journal of the American Society for Mass Spectrometry · 2016Article
- Quantification of Human Kallikrein-Related Peptidases in Biological Fluids by Multiplatform Targeted Mass Spectrometry Assays.Molecular & cellular proteomics : MCP · 2016Article
Corrections and comments
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Authors and funding
5 authors.
Funding
Abstract
Liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM-MS) of plasma that has been depleted of abundant proteins and fractionated at the peptide level into six to eight fractions is a proven method for quantifying proteins present at low nanogram-per-milliliter levels. A drawback of fraction-MRM is the increased analysis time due to the generation of multiple fractions per biological sample. We now report that the use of heated, long, fused silica columns (>30 cm) packed with 1.9 μm of packing material can reduce or eliminate the need for fractionation prior to LC-MRM-MS without a significant loss of sensitivity or precision relative to fraction-MRM. We empirically determined the optimal column length, temperature, gradient duration, and sample load for such assays and used these conditions to study detection sensitivity and assay precision. In addition to increased peak capacity, longer columns packed with smaller beads tolerated a 4- to 6-fold increase in analyte load without a loss of robustness or reproducibility. The longer columns also provided a 4-fold improvement in median limit-of-quantitation values with increased assay precision relative to the standard 12 cm columns packed with 3 μm material. Overall, the optimized chromatography provided an approximately 3-fold increase in analysis throughput with excellent robustness and less than a 2-fold reduction in quantitative sensitivity relative to fraction-MRM. The value of the system for increased multiplexing was demonstrated by the ability to configure an 800-plex MRM-MS assay, run in a single analysis, comprising 2400 transitions with retention time scheduling to monitor 400 unlabeled and heavy labeled peptide pairs.
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Registered trials
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