Improved Electrospray Ionization Efficiency Compensates for Diminished Chromatographic Resolution and Enables Proteomics Analysis of Tyrosine Signaling in Embryonic Stem Cells
2009; American Chemical Society; Volume: 81; Issue: 9 Linguagem: Inglês
10.1021/ac802720e
ISSN1520-6882
AutoresScott B. Ficarro, Yi Zhang, Yu Lu, Ahmadali R. Moghimi, Manor Askenazi, Elzbieta Hyatt, Eric Smith, Leah Boyer, Thorsten M. Schlaeger, Chance John Luckey, Jarrod A. Marto,
Tópico(s)Microfluidic and Capillary Electrophoresis Applications
ResumoCharacterization of signaling pathways in embryonic stem cells is a prerequisite for future application of these cells to treat human disease and other disorders. Identification of tyrosine signaling cascades is of particular interest but is complicated by the relatively low levels of tyrosine phosphorylation in embryonic stem cells. These hurdles correlate with the primary limitations of mass spectrometry-based proteomics; namely, poor detection limit and dynamic range. To overcome these obstacles, we fabricated miniaturized LC-electrospray assemblies that provided ∼15-fold improvement in LC-MS performance. Significantly, our characterization data demonstrate that electrospray ionization efficiency compensates for diminished chromatographic performance at effluent flow rates below Van Deemter minima. Use of these assemblies facilitated quantitative proteomics-based analysis of tyrosine signaling cascades in embryonic stem cells. Our results suggest that a renewed focus on miniaturized LC coupled to ultralow flow electrospray will provide a viable path for proteomic analysis of primary cells and rare post-translational modifications.
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