High pH reversed-phase chromatography with fraction concatenation for 2D proteomic analysis
Orthogonal high-resolution separations are critical for attaining improved analytical dynamic ranges of proteome measurements. Concatenated high pH reversed phase liquid chromatography affords better separations than the strong cation exchange conventionally applied for two-dimensional shotgun proteomic analysis. For example, concatenated high pH reversed phase liquid chromatography increased identification coverage for peptides (e.g., by 1.8-fold) and proteins (e.g., by 1.6-fold) in shotgun proteomics analyses of a digested human protein sample. Additional advantages of concatenated high pH RPLC include improved protein sequence coverage, simplified sample processing, and reduced sample losses, making this an attractive first dimension separation strategy for two-dimensional proteomics analyses.
- Research Organization:
- Pacific Northwest National Laboratory (PNNL), Richland, WA (US), Environmental Molecular Sciences Laboratory (EMSL)
- Sponsoring Organization:
- USDOE
- DOE Contract Number:
- AC05-76RL01830
- OSTI ID:
- 1038643
- Report Number(s):
- PNNL-SA-83356; 40072; 400412000
- Journal Information:
- Expert Review of Proteomics, 9(2):129-134, Journal Name: Expert Review of Proteomics, 9(2):129-134 Journal Issue: 2 Vol. 9
- Country of Publication:
- United States
- Language:
- English
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