Differential recovery of peptides from sample tubes and the reproducibility of quantitative proteomic data
Differential recovery of peptides from sample tubes and the reproducibility of quantitative proteomic data
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DOI：
10.1021/pr070294o
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发表时间：
2007-11-01
影响因子：
4.4
通讯作者：
Hook, Vivian
中科院分区：
文献类型：
--
作者：
Bark, Steven J.;Hook, Vivian
由于非特异性吸附导致的肽的差异回收会严重损害液相色谱-质谱法（LC-MS）用于肽分析的蛋白质组学数据的重现性和质量。本研究表明，在不同样品管中储存时，BSA胰蛋白酶解产物衍生肽的LC-MS数据的重现性和定量存在较大差异。值得注意的是，我们表明，高度改善的一致性和较低的错误，在重复测量中的BSA胰蛋白酶肽定量实现与低保留管相比，定期eppendorf管。此外，在两种类型的储存管中观察到通过LC-MS检测到的肽的定性差异。这些结果说明了仔细评价储存容器和条件的必要性，以最大限度地减少LC-MS实验样品质量的变异性。
Differential recovery of peptides due to nonspecific adsorption can seriously compromise reproducibility and quality of proteomic data for pepticle analyses by liquid chromatography-mass spectrometry (LC-MS). This study demonstrates large variations in reproducibility and quantitation of LC-MS data for peptides derived from tryptic digests of BSA upon storage in different sample tubes. Notably, we show that highly improved consistency and lower errors in quantitation of BSA tryptic peptides in replicate measurements is achieved with low-retention tubes compared to regular eppendorf tubes. Furthermore, qualitative differences in peptides detected by LC-MS were observed in the two types of storage tubes. These results illustrate the necessity for careful evaluation of storage vessels and conditions to minimize variability in sample quality for LC-MS experiments.