ArticleJournal of proteome research2013
Hydrophilic strong anion exchange (hSAX) chromatography for highly orthogonal peptide separation of complex proteomes.
Article in Journal of proteome research, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 35 papers.
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Who cites it
35 citing papers in PubMed.
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- Enrichment Workflow for Pyrophosphoproteome Analysis.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Extensive protein pyrophosphorylation revealed in human cell lines.Nature chemical biology · 2024Article
- Impact of sacubitril/valsartan on cardiac and systemic hypoxia in chronic heart failure.iScience · 2024Article
- Proteomic and transcriptomic profiling of aerial organ development in Arabidopsis.Scientific data · 2020Article
- A mass spectrometry-based proteome map of drug action in lung cancer cell lines.Nature chemical biology · 2020Article
- Fractionation of Enriched Phosphopeptides Using pH/Acetonitrile-Gradient-Reversed-Phase Microcolumn Separation in Combination with LC-MS/MS Analysis.International journal of molecular sciences · 2020Article
- The type VI secretion system deploys antifungal effectors against microbial competitors.Nature microbiology · 2018Article
- LRRK2 is a negative regulator ofThe EMBO journal · 2018Article
- Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib.The EMBO journal · 2018Article
- Peptide Retention in Hydrophilic Strong Anion Exchange Chromatography Is Driven by Charged and Aromatic Residues.Analytical chemistry · 2018Article
- Improved mass spectrometric detection of acidic peptides by variations in the functional group pKThe Analyst · 2018Article
- Proteome-wide analysis of protein abundance and turnover remodelling during oncogenic transformation of human breast epithelial cells.Wellcome open research · 2018Article
- 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
- Expanding Proteome Coverage with CHarge Ordered Parallel Ion aNalysis (CHOPIN) Combined with Broad Specificity Proteolysis.Journal of proteome research · 2017Article
- Towards comprehensive and quantitative proteomics for diagnosis and therapy of human disease.Proteomics · 2017Review
- Towards Understanding Male Infertility After Spinal Cord Injury Using Quantitative Proteomics.Molecular & cellular proteomics : MCP · 2016Article
- IFNs Modify the Proteome of Legionella-Containing Vacuoles and Restrict Infection Via IRG1-Derived Itaconic Acid.PLoS pathogens · 2016Article
- Enhanced snoMEN Vectors Facilitate Establishment of GFP-HIF-1α Protein Replacement Human Cell Lines.PloS one · 2016Article
- Evaluating the use of HILIC in large-scale, multi dimensional proteomics: Horses for courses?International journal of mass spectrometry · 2015Article
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5 authors.
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Abstract
Due to its compatibility and orthogonality to reversed phase (RP) liquid chromatography (LC) separation, ion exchange chromatography, and mainly strong cation exchange (SCX), has often been the first choice in multidimensional LC experiments in proteomics. Here, we have tested the ability of three strong anion exchanger (SAX) columns differing in their hydrophobicity to fractionate RAW264.7 macrophage cell lysate. IonPac AS24, a strong anion exchange material with ultralow hydrophobicity, demonstrated to be superior to other materials by fractionation and separation of tryptic peptides from both a mixture of 6 proteins as well as mouse cell lysate. The chromatography displayed very high orthogonality and high robustness depending on the hydrophilicity of column chemistry, which we termed hydrophilic strong anion exchange (hSAX). Mass spectrometry analysis of 34 SAX fractions from RAW264.7 macrophage cell lysate digest resulted in an identification of 9469 unique proteins and 126318 distinct peptides in one week of instrument time. Moreover, when compared to an optimized high pH/low pH RP separation approach, the method presented here raised the identification of proteins and peptides by 10 and 28%, respectively. This novel hSAX approach provides robust, reproducible, and highly orthogonal separation of complex protein digest samples for deep coverage proteome analysis.
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