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ERLIC-based strategies for combined quantitative proteome and phosphoproteome analysis with limited sample amounts

ABSTRACT: The study aims the development of a sensitive strategy for combined quantitative proteomics and phosphoproteomics. 3 ERLIC-based strategies, applicable for low-sample-amount experiments (~150 µg per condition), were evaluated and compared . Three samle sets have been uploaded. 16 fractions ERLIC-1-SCX/RP (2 replicates) for phosphopeptide enrichment, 8 fractions SCX-EL (2 replicates), 12 fractions ERLIC-2 (2 replicates). The following 3 strategies were compared: A 2D strategy -> sample set ERLIC-1-SCX/RP. A pseudo-3Dstrategy -> sample sets ERLIC-1-SCX/RP + SCX-EL and a 3D strategy -> sample sets ERLIC-1-SCX/RP + ERLIC-2

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Cell Culture, Fibroblast

SUBMITTER: Stefan Loroch

LAB HEAD: Albert Sickmann

PROVIDER: PXD001241 | Pride | 2015-02-25

REPOSITORIES: Pride

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Multidimensional electrostatic repulsion-hydrophilic interaction chromatography (ERLIC) for quantitative analysis of the proteome and phosphoproteome in clinical and biomedical research.

Loroch Stefan S Schommartz Tim T Brune Wolfram W Zahedi René Peiman RP Sickmann Albert A

Biochimica et biophysica acta 20150122 5

Quantitative proteomics and phosphoproteomics have become key disciplines in understanding cellular processes. Fundamental research can be done using cell culture providing researchers with virtually infinite sample amounts. In contrast, clinical, pre-clinical and biomedical research is often restricted to minute sample amounts and requires an efficient analysis with only micrograms of protein. To address this issue, we generated a highly sensitive workflow for combined LC-MS-based quantitative ...[more]

PMID: 25619855

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