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肽的储存:你的投资是否获得了最佳回报?确定蛋白质组学样品的最佳储存条件。

Peptide storage: are you getting the best return on your investment? Defining optimal storage conditions for proteomics samples.

作者信息

Kraut Alexandra, Marcellin Marlène, Adrait Annie, Kuhn Lauriane, Louwagie Mathilde, Kieffer-Jaquinod Sylvie, Lebert Dorothée, Masselon Christophe D, Dupuis Alain, Bruley Christophe, Jaquinod Michel, Garin Jérôme, Gallagher-Gambarelli Maighread

机构信息

CEA, DSV, iRTSV, Laboratoire d'Etude de la Dynamique des Protéomes, INSERM, U880, Université Joseph Fourier, Grenoble, F-38054, France.

出版信息

J Proteome Res. 2009 Jul;8(7):3778-85. doi: 10.1021/pr900095u.

Abstract

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.

摘要

为了符合当前的蛋白质组学指南,通常有必要对相同的肽样品进行多次分析。在分析之间,样品必须以能够保存其固有特性的方式进行储存,既不损失肽也不损失信号强度。本文介绍了两项旨在确定分析之间肽样品最佳储存条件的研究。采用无标记策略,在28天的时间内,对三种不同的容器:标准塑料管、玻璃管和低吸附塑料管中的肽保存情况进行了比较。这项研究的结果表明,在所研究的时间段内,标准塑料管不适用于肽的储存。发现玻璃管的性能优于标准塑料,但使用低吸附塑料管可实现最佳的肽回收率。储存后回收率较差的肽主要具有疏水性。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。这项研究能够在同一液相色谱-质谱运行中准确比较两种管型之间的肽回收率。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率都是最佳的。

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