Improved Electrospray Ionization Efficiency Compensates for Diminished Chromatographic Resolution and Enables Proteomics Analysis of Tyrosine Signaling in Embryonic Stem Cells

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• 作者：Scott B. Ficarro ; Yi Zhang ; Yu Lu ; Ahmadali R. Moghimi ; Manor Askenazi ; Elzbieta Hyatt ; Eric D. Smith ; Leah Boyer ; Thorsten M. Schlaeger ; C. John Luckey ; Jarrod A. Marto
• 刊名：Analytical Chemistry
• 出版年：2009
• 出版时间：May 1, 2009
• 年：2009
• 卷：81
• 期：9
• 页码：3440-3447
• 全文大小：417K
• 年卷期：v.81,no.9(May 1, 2009)
• ISSN：1520-6882

文摘

Characterization 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.