Effects of storage time and exogenous protease inhibitors on plasma protein levels
Effects of storage time and exogenous protease inhibitors on plasma protein levels
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DOI：
10.1309/3wm7xj7rd8bclnkx
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发表时间：
2006-08-01
影响因子：
3.5
通讯作者：
Stroncek, David F.
中科院分区：
文献类型：
--
作者：
Ayache, Saleh;Panelli, Monica;Stroncek, David F.
血浆生物标志物分析需要完整无偏倚的起始材料。我们分析了蛋白酶抑制剂鸡尾酒和预处理储存对血浆蛋白质谱的影响。立即和室温储存2小时后制备从12名健康受试者中采集的含和不含蛋白酶抑制剂的血浆。通过捕获99种趋化因子和细胞因子的多重酶联免疫吸附测定来分析样品。无监督分层分析将样本聚类为4组：一组主要由立即处理的样本组成，另一组由2小时后处理的样本组成，其余两组由立即和2小时样本组成。混合组和即刻组蛋白质谱相似，但是，在即刻组和2小时组样本中，37个因子的水平显著不同：2小时后所有因子均更高。储存期间蛋白质水平的显著变化与蛋白酶抑制剂无关，可能是由于白细胞和血小板产生和/或释放细胞因子所致。
Plasma biomarker analysis requires intact unbiased starting material. We analyzed the effects on plasma protein profiles of protease inhibitor cocktails and preprocessing storage. Plasma from 12 healthy subjects collected with and without protease inhibitors was prepared immediately and after 2 hours of room temperature storage. The samples were analyzed by a multiplexed enzyme-linked immunosorbent assay that captured 99 chemokines and cytokines. Unsupervised hierarchical analysis clustered the samples into 4 groups: one composed predominantly of samples processed immediately, another of samples processed after 2 hours, and the remaining two a mix of immediate and 2-hour samples. The mixed and immediate group protein profiles were similar However, among the immediate and 2-hour samples, the levels of 37 factors differed significantly: all were greater after 2 hours. The dramatic changes in protein levels during storage were independent of protease inhibitors and are likely due to cytokine production and/or release by leukocytes and platelets.