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

• DOI: 10.1586/epr.12.15
• Publication Date: 2012-04-02T10:31:42Z
• Authors: Feng Yang; Yufeng Shen; David G Camp; Richard D Smith
• Keywords: Shotgun proteomics; Concatenation (mathematics); Fraction (chemistry)

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 applied 2D analysis. For example, concatenated high-pH RPLC increased identification of peptides (by 1.8-fold) proteins 1.6-fold) shotgun proteomics analyses a digested human sample. Additional advantages with concatenation include sequence coverage, simplified sample processing reduced losses, making this an attractive alternative to conjunction second-dimension low-pH analyses.