Online Nanoscale ERLIC-MS Outperforms RPLC-MS for Shotgun Proteomics in Complex Mixtures
Online Nanoscale ERLIC-MS Outperforms RPLC-MS for Shotgun Proteomics in Complex Mixtures
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DOI：
10.1021/pr300638n
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发表时间：
2012-10-01
影响因子：
4.4
通讯作者：
Griffin, Timothy J.
中科院分区：
文献类型：
--
作者：
de Jong, Ebbing P.;Griffin, Timothy J.
我们已经探索了使用静电排斥亲水相互作用色谱法（ERLIC）作为替代的金标准，在鸟枪蛋白质组学：反相（RP）LC在线ESI-MS/MS。样品溶解和初始梯度条件进行了优化，以达到肽的溶解度和最大肽保留之间的平衡时，使用高有机溶剂浓度的移动的相。在线ERLIC-MS表现出57%的总肽鉴定相比，RP-MS。我们研究了这种改进的性能的机制，发现它源于ERLIC的倾向，保留较长的肽，可以确定更大的信心。在线纳米ERLIC-MS提供了一个强大的新工具，以提高MS为基础的鸟枪蛋白质组学在广泛的应用。
We have explored the use of electrostatic repulsion hydrophilic interaction chromatography (ERLIC) as an alternative to the gold-standard in shotgun proteomics: reversed-phase (RP) LC for online ESI-MS/MS. Conditions for sample solubilization and initial gradient conditions were optimized to strike a balance between peptide solubility and maximum peptide retention when using mobile phase with high organic solvent concentration. Online ERLIC-MS demonstrated a 57% increase in total peptide identifications compared to RP-MS. We examined the mechanism of this improved performance and found that it stems from ERLIC's propensity to retain longer peptides, which can be identified with greater confidence. Online nanoscale ERLIC-MS provides a powerful new tool for enhancing MS-based shotgun proteomic in a broad range of applications.