| Hydrophilic Strong Anion Exchange (hSAX) Chromatography for Highly Orthogonal Peptide Separation of Complex Proteomes. | |
| | |
MedLine Citation:
|
PMID: 23294059 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
|
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. |
| | |
Authors:
|
Maria Stella Ritorto; Ken Cook; Kshitiz Tyagi; Patrick G A Pedrioli; Matthias Trost |
Related Documents
:
|
12622389 - Three-electrode electrochemical detector and platinum film decoupler integrated with a ... 6526899 - Determination of oxalic acid in biological matrices by liquid chromatography with amper... 18962539 - Determination of mercaptopyrimidines with copper(ii) in acetonitrile. 15042169 - Simultaneous quantification of analytes in quaternary mixtures using dual pulse stairca... 15571339 - Electrochemical detection method for nonelectroactive and electroactive analytes in mic... 16867629 - A poly(vinylchloride) membrane electrode for determination of phenytoin in pharmaceutic... 16915829 - Determination of aristolochic acid in botanicals and dietary supplements by liquid chro... 19997879 - Determination of equilibrium association constants of ligand-dna complexes by electrosp... 6819409 - Chemical analysis of human blood for assessment of environmental exposure to semivolati... |
Publication Detail:
|
Type: JOURNAL ARTICLE Date: 2013-1-7 |
Journal Detail:
|
Title: Journal of proteome research Volume: - ISSN: 1535-3907 ISO Abbreviation: J. Proteome Res. Publication Date: 2013 Jan |
Date Detail:
|
Created Date: 2013-1-8 Completed Date: - Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 101128775 Medline TA: J Proteome Res Country: - |
Other Details:
|
Languages: ENG Pagination: - Citation Subset: - |
Affiliation:
|
MRC Protein Phosphorylation and Ubiquitylation Unit, University of Dundee , Dundee, United Kingdom. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
|
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Clinical impact of the publication of S3 guidelines for intensive care in cardiac surgery patients i...
Next Document: The Revolution and Evolution of Shotgun Proteomics for Large-Scale Proteome Analysis.