Bark Steven J, Hook Vivian
The Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of California, San Diego, 9500 Gilman Drive, MC 0744, La Jolla, California 92093-0744, USA.
J Proteome Res. 2007 Nov;6(11):4511-6. doi: 10.1021/pr070294o. Epub 2007 Sep 12.
Differential recovery of peptides due to nonspecific adsorption can seriously compromise reproducibility and quality of proteomic data for peptide 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.
由于非特异性吸附导致的肽段回收率差异会严重影响液相色谱-质谱联用（LC-MS）肽段分析蛋白质组学数据的重现性和质量。本研究表明，牛血清白蛋白（BSA）胰蛋白酶消化产物衍生的肽段在不同样品管中储存后，LC-MS数据的重现性和定量存在很大差异。值得注意的是，我们发现与常规的艾本德管相比，使用低保留管可在重复测量中实现更高的一致性和更低的BSA胰蛋白酶肽段定量误差。此外，在两种类型的储存管中观察到LC-MS检测到的肽段存在定性差异。这些结果表明，有必要仔细评估储存容器和条件，以尽量减少LC-MS实验中样品质量的变异性。
