Peptide Storage: Are You Getting the Best Return on Your Investment? Defining Optimal Storage Conditions for Proteomics Samples
Peptide Storage: Are You Getting the Best Return on Your Investment? Defining Optimal Storage Conditions for Proteomics Samples
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DOI：
10.1021/pr900095u
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发表时间：
2009-07-01
影响因子：
4.4
通讯作者：
Gallagher-Gambarelli, Maighread
中科院分区：
文献类型：
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作者：
Kraut, Alexandra;Marcellin, Marlene;Gallagher-Gambarelli, Maighread
为了符合当前的蛋白质组学指南，通常需要对相同的肽样品进行多次分析。在两次分析之间，样品的存储方式必须能够保留其内在特性，而不损失肽或信号强度。本文介绍了两项旨在确定分析之间肽样品的最佳储存条件的研究。通过使用无标记策略，在 28 天的时间内对三种不同接受者的肽保守性进行了比较：标准塑料管、玻璃管和低吸附塑料管。这项研究的结果表明，标准塑料管不适合在研究期间储存肽。研究发现玻璃管的性能优于标准塑料管，但使用低吸附塑料管可实现最佳的肽回收率。储存后表现出较差恢复性的肽本质上主要是疏水性的。使用同位素标记的蛋白质进一步研究了玻璃管和低吸附塑料管之间肽回收率的差异。这项研究允许在同一 LC-MS 运行中准确比较两种管类型之间的肽回收率。无标记研究的结果得到了证实。此外，还可以证明，无论储存的肽浓度如何，低吸附塑料管中的肽回收都是最佳的。
To comply with current proteomics guidelines, it is often necessary to analyze the same peptide samples several times. Between analyses, the sample must be stored in such a way as to conserve its intrinsic properties, without losing either peptides or signal intensity. This article describes two studies designed to define the optimal storage conditions for peptide samples between analyses. With the use of a label-free strategy, peptide conservation was compared over a 28-day period in three different recipients: standard plastic tubes, glass tubes, and low-adsorption plastic tubes. The results of this study showed that standard plastic tubes are unsuitable for peptide storage over the period studied. Glass tubes were found to perform better than standard plastic, but optimal peptide recovery was achieved using low-adsorption plastic tubes. The peptides showing poor recovery following storage were mainly hydrophobic in nature. The differences in peptide recovery between glass and low-adsorption plastic tubes were further studied using isotopically labeled proteins. This study allowed accurate comparison of peptide recovery between the two tube types within the same LC-MS run. The results of the label-free study were confirmed. Further, it was possible to demonstrate that peptide recovery in low-adsorption plastic tubes was optimal whatever the peptide concentration stored.