title: Fully automated isotopic dimethyl labeling and phosphopeptide enrichment using a microfluidic HPLC phosphochip.
abstract: 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 (TiO(2)) 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.
authors.affiliation: Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
authors.forename: Ayse Nur
authors.initials: AN
authors.lastname: Polat
authors.forename: Karsten
authors.initials: K
authors.lastname: Kraiczek
authors.forename: Albert J R
authors.initials: AJ
authors.lastname: Heck
authors.forename: Reinout
authors.initials: R
authors.lastname: Raijmakers
authors.forename: Shabaz
authors.initials: S
authors.lastname: Mohammed
mesh_terms: D001331:Automation; D002851:Chromatography, High Pressure Liquid / Q000592:standards*; D008401:Gas Chromatography-Mass Spectrometry; D007553:Isotope Labeling / Q000379:methods*; D044085:Microfluidics / Q000592:standards*; D010748:Phosphopeptides / Q000737:chemistry*; D015203:Reproducibility of Results; D014025:Titanium / Q000737:chemistry
chemical_list: D010748:Phosphopeptides; C009495:titanium dioxide; D014025:Titanium