Evaluation of blood collection tubes using selected reaction monitoring MS: Implications for proteomic biomarker studies
Evaluation of blood collection tubes using selected reaction monitoring MS: Implications for proteomic biomarker studies
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DOI：
10.1002/pmic.200900517
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发表时间：
2010-05-01
期刊：
影响因子：
3.4
通讯作者：
Molloy, Mark P.
中科院分区：
文献类型：
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作者：
Randall, Sarah A.;McKay, Matthew J.;Molloy, Mark P.
当前蛋白质组学研究的一个重点是血浆蛋白生物标志物的验证。血液采集过程本身对定量生物标志物测定的准确性和可重复性至关重要。我们开发了选择性反应监测（SRM）方法来分析13种丰富的血浆蛋白，并评估了三种不同的采血管对这些蛋白水平的影响。我们还评估了通过评估这些蛋白质的恢复来分析血浆样本所需时间的含义。我们发现SRM检测到一些蛋白质水平的微小差异，这可以归因于收集管类型。与传统的收集管相比，在含有蛋白酶抑制剂的管中收集的蛋白质，18项测定中有12项的平均回收率更高。对于其中的五种测定，差异回收率具有统计学意义。将冻融样品的质谱分析延迟1小时，这些分析物的回收率大大降低；然而，管型差异仅在基线时间点明显。最后，我们在7个健康个体的队列中评估了这些蛋白质循环水平的自然变化。本研究为研究人员考虑采集血液进行蛋白质生物标志物研究提供了有用的信息。
An emphasis of current proteomic research is the validation of plasma protein biomarkers. The process of blood collection itself is critical to the accuracy and reproducibility of quantitative biomarker assays. We have developed selected reaction monitoring (SRM) assays to analyse thirteen abundant plasma proteins and evaluated the impact of three different blood collection tubes on the levels of these proteins. We also assessed the implications of the time taken to analyse plasma samples by evaluating the recovery of these proteins. We showed that SRM detects minor differences in the levels of some proteins which can be attributed to collection tube type. The average recovery for 12 of 18 assays was higher for proteins that were collected in tubes containing protease inhibitors compared to conventional collection tubes. For five of the assays, the differential recovery was statistically significant. Delaying MS analysis of a freeze-thawed sample for 1 hour showed greatly reduced recovery of these analytes; however differences attributed to tube type were only evident at the baseline timepoint. Finally, we assessed the natural variation of circulating levels of these proteins in a cohort of seven healthy individuals. This study provides useful information for researchers contemplating blood collection for undertaking protein biomarker studies.