Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 404, Issue 8, Pages 2507-2512Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-012-6395-7
Keywords
HPLC chip; Microfluidics; Dimethyl labeling; Phosphorylation; Quantification; TiO2
Funding
- PRIMEXS project [262067]
- European Union
- Netherlands Proteomics Centre, embedded in the Netherlands Genomics Initiative, The Netherlands Organization for Scientific Research (NWO)
- VIDI [700.10.429]
- Agilent Technologies Foundation
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Quantitative detection of phosphorylation levels is challenging and requires an expertise in both stable isotope labeling as well as enrichment of phosphorylated peptides. Recently, a microfluidic device incorporating a nanoliter flow rate reversed phase column as well as a titania (TiO2) enrichment column was released. This HPLC phosphochip allows excellent recovery and separation of phosphorylated peptides in a robust and reproducible manner with little user intervention. In this work, we have extended the abilities of this chip by defining the conditions required for on-chip stable isotope dimethyl labeling allowing for automated quantitation. The resulting approach will make quantitative phosphoproteomics more accessible.
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