ArticleAnalytical chemistry2011
Online nanoflow reversed phase-strong anion exchange-reversed phase liquid chromatography-tandem mass spectrometry platform for efficient and in-depth proteome sequence analysis of complex organisms.
Article in Analytical chemistry, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 64 citations in OpenAlex.
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- Identification and validation of selective deubiquitinase inhibitors.Cell chemical biology · 2021Article
- Latest Trends on the Future of Three-Dimensional Separations in Chromatography.Chemical reviews · 2021Review
- Review of Three-Dimensional Liquid Chromatography Platforms for Bottom-Up Proteomics.International journal of molecular sciences · 2020Review
- Leveraging Compound Promiscuity to Identify Targetable Cysteines within the Kinome.Cell chemical biology · 2019Article
- In vitro analysis of RNA polymerase II elongation complex dynamics.Genes & development · 2019Article
- A Chemoproteomic Strategy for Direct and Proteome-Wide Covalent Inhibitor Target-Site Identification.Journal of the American Chemical Society · 2019Article
- High-throughput metabolomics used to identify potential therapeutic targets of Guizhi Fuling Wan against endometriosis of cold coagulation and blood stasis.RSC advances · 2018Article
- Salivary Protein Roles in Oral Health and as Predictors of Caries Risk.Open life sciences · 2018Article
- Advances in microscale separations towards nanoproteomics applications.Journal of chromatography. A · 2017Review
- Highly reproducible improved label-free quantitative analysis of cellular phosphoproteome by optimization of LC-MS/MS gradient and analytical column construction.Journal of proteomics · 2017Article
- Phosphoproteomic profiling of mouse primary HSPCs reveals new regulators of HSPC mobilization.Blood · 2016Article
- Review
- Off-Line Multidimensional Liquid Chromatography and Auto Sampling Result in Sample Loss in LC/LC-MS/MS.Journal of proteome research · 2014Article
- Hydrophilic strong anion exchange (hSAX) chromatography for highly orthogonal peptide separation of complex proteomes.Journal of proteome research · 2013Article
- Protein analysis by shotgun/bottom-up proteomics.Chemical reviews · 2013Review
- Genome-scale proteome quantification by DEEP SEQ mass spectrometry.Nature communications · 2013Article
- Proteomic analysis demonstrates activator- and chromatin-specific recruitment to promoters.The Journal of biological chemistry · 2012Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
The dynamic range of protein expression in complex organisms coupled with the stochastic nature of discovery-driven tandem mass spectrometry (MS/MS) analysis continues to impede comprehensive sequence analysis and often provides only limited information for low-abundance proteins. High-performance fractionation of proteins or peptides prior to mass spectrometry analysis can mitigate these effects, though achieving an optimal combination of automation, reproducibility, separation peak capacity, and sample yield remains a significant challenge. Here we demonstrate an automated nanoflow 3-D liquid chromatography (LC)-MS/MS platform based on high-pH reversed phase (RP), strong anion exchange (SAX), and low-pH reversed phase (RP) separation stages for analysis of complex proteomes. We observed that RP-SAX-RP outperformed RP-RP for analysis of tryptic peptides derived from Escherichia coli and enabled identification of proteins present at a level of 50 copies per cell in Saccharomyces cerevisiae, corresponding to an estimated detection limit of 500 amol, from 40 μg of total lysate on a low-resolution 3-D ion trap mass spectrometer. A similar study performed on a LTQ-Orbitrap yielded over 4000 unique proteins from 5 μg of total yeast lysate analyzed in a single, 101 fraction RP-SAX-RP LC-MS/MS acquisition, providing an estimated detection limit of 65 amol for proteins expressed at 50 copies per cell.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.