Systematical evaluation of the effects of sample collection procedures on low-molecular-weight serum/plasma proteome profiling
Systematical evaluation of the effects of sample collection procedures on low-molecular-weight serum/plasma proteome profiling
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DOI：
10.1002/pmic.200500535
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发表时间：
2006-05-01
期刊：
影响因子：
3.4
通讯作者：
Lee, Hai-Lun
中科院分区：
文献类型：
--
作者：
Hsieh, Sen-Yung;Chen, Ren-Kung;Lee, Hai-Lun
血液是生物标志物发现的理想来源。然而，几乎没有做什么来解决的采样，处理和储存程序对血清/血浆蛋白质组的影响。我们使用基于磁珠的MALDI-TOF MS系统地评估每个程序对低分子量血清/血浆蛋白质组分析的基础上的整个光谱的影响。我们发现，采样程序，包括采血管和抗凝剂的选择，在离心前的点时间或时滞的变化，和溶血，显示显着的蛋白质组的影响。此外，血清和血浆是相互不相容的蛋白质组比较。相比之下，过夜禁食，处理程序，包括离心速度（1500 x g与3000 x g）或时间（15分钟对30分钟）和储存条件，如在4 ℃或25 ℃下长达24小时或在-80 ℃下长达3个月，以及重复冷冻/解冻长达10个循环，根据我们对大约100个峰的分析，对蛋白质组的影响相对较小。我们得出结论，低分子量血清/血浆蛋白质组的多样性受采样，处理和储存的变化，从采样程序的变化。因此，我们建议有必要标准化的采样程序，蛋白质组比较和生物标志物的发现。
Blood is an ideal source for biomarker discovery. However, little has been done to address the effects of sampling, handling and storage procedures on serum/plasma proteomes. We used magnetic bead-based MALDI-TOF MS to systematically evaluate the influence of each procedure on low-molecular-weight serum/plasma proteome profiling on the basis of the whole spectra. We found that sampling procedures, including the selection of blood collection tubes and anticoagulants, variations in dotting time or time lag before centrifugation, and hemolysis, displayed significant effects on the proteomes. Moreover, serum and plasma were mutually incompatible for proteome comparison. By contrast, overnight fasting, handling procedures, including centrifugation speeds (1500 x g vs. 3000 x g) or time (15 min vs. 30 min), and storage conditions, such as at 4 degrees C or 25 degrees C for up to 24 h or at -80 degrees C for up to 3 months, and repeated freeze/thaw of up to ten cycles, had relatively minor effects on the proteomes based upon our analysis of about 100 peaks. We concluded that low-molecular-weight serum/plasma proteomes were diversely affected by sampling, handling and storage with most change from variations of sampling procedures. We therefore suggest the necessity of standardizing sampling procedure for proteome comparison and biomarker discovery.