Literature DB >> 22462785

High-pH reversed-phase chromatography with fraction concatenation for 2D proteomic analysis.

Feng Yang1, Yufeng Shen, David G Camp, Richard D Smith.   

Abstract

Orthogonal high-resolution separations are critical for attaining improved analytical dynamic range and protein coverage in proteomic measurements. High-pH reversed-phase liquid chromatography (RPLC), followed by fraction concatenation, affords better peptide analysis than conventional strong cation-exchange chromatography applied for 2D proteomic analysis. For example, concatenated high-pH RPLC increased identification of peptides (by 1.8-fold) and proteins (by 1.6-fold) in shotgun proteomics analyses of a digested human protein sample. Additional advantages of high-pH RPLC with fraction concatenation include improved protein sequence coverage, simplified sample processing and reduced sample losses, making this an attractive alternative to strong cation-exchange chromatography in conjunction with second-dimension low-pH RPLC for 2D proteomics analyses.

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Year:  2012        PMID： 22462785      PMCID： PMC3337716          DOI： 10.1586/epr.12.15

Source DB:  PubMed          Journal:  Expert Rev Proteomics        ISSN： 1478-9450            Impact factor:   3.940


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