Definitive screening design enables optimization of LC-ESI-MS/MS parameters in proteomics
Definitive screening design enables optimization of LC-ESI-MS/MS parameters in proteomics
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DOI：
10.1080/09168451.2017.1391685
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发表时间：
2017-01-01
影响因子：
1.6
通讯作者：
Ueda, Mitsuyoshi
中科院分区：
文献类型：
--
作者：
Aburaya, Shunsuke;Aoki, Wataru;Ueda, Mitsuyoshi
在蛋白质组学中，超过100，000种肽是从人类细胞裂解物的消化中产生的。蛋白质组样品具有广泛的蛋白质丰度动态范围;因此，优化LC-ESI-MS/MS的各种参数以全面鉴定这些肽至关重要。然而，LC-ESI-MS/MS分析有许多参数。在这项研究中，我们采用确定性筛选设计，同时优化整体毛细管LC-ESI-MS/MS操作中的14个参数，以增加识别的蛋白质的数量和/或MS 1的平均峰面积。同时优化使LC和MS之间的双因素相互作用的测定。最后，我们发现了两个参数集的整体毛细管LC-ESI-MS/MS，增加了8.1%或MS 1的平均峰面积的67%，确定的蛋白质的数量。确定的筛选设计将是非常有用的高通量分析的最佳参数设置在LC-ESI-MS/MS系统。
In proteomics, more than 100,000 peptides are generated from the digestion of human cell lysates. Proteome samples have a broad dynamic range in protein abundance; therefore, it is critical to optimize various parameters of LC-ESI-MS/MS to comprehensively identify these peptides. However, there are many parameters for LC-ESI-MS/MS analysis. In this study, we applied definitive screening design to simultaneously optimize 14 parameters in the operation of monolithic capillary LC-ESI-MS/MS to increase the number of identified proteins and/or the average peak area of MS1. The simultaneous optimization enabled the determination of two-factor interactions between LC and MS. Finally, we found two parameter sets of monolithic capillary LC-ESI-MS/MS that increased the number of identified proteins by 8.1% or the average peak area of MS1 by 67%. The definitive screening design would be highly useful for high-throughput analysis of the best parameter set in LC-ESI-MS/MS systems.