The purpose of the present study was to assess the differences between the human serum and plasma proteomes, and the stability of human plasma proteins under different storage conditions following blood collection, by means of SWATH-MS analysis. When we compared plasma and serum prepared immediately after blood sampling, 95.5% of 176 quantified proteins differed by less than 1.5-fold. When we compared plasma samples prepared by centrifugation after storage of blood at room temperature for 0, 15 or 30 min, or under refrigeration at 0-5 °C for 1, 4 or 8 h, no protein showed a significant change (q < 0.05) that amounted to 1.5-fold or more, except hemoglobins. Those proteins were greatly increased in a single sample at 8 h, probably due to hemolysis. Comparison of data from the same samples indicates that the blood proteome is more stable than the blood metabolome. The present results suggest that most components of the proteome are essentially the same in plasma and serum, and are stable under the storage conditions examined in the present study. However, it may be important to pay attention to the extent of coagulation, and levels of platelet and hemolysis-related proteins. SIGNIFICANCE: Pre-analytical processing and storage conditions after blood collection are expected to influence the blood proteome. Therefore, we investigated differences in the proteome between human serum and plasma, as well as the stability of human plasma proteins under different storage conditions: at room temperature for 0-30 min, or at 0-5 °C for 1-8 h, which may reflect the clinical situation of blood collection. Proteomics analysis with SWATH-MS identified 342 proteins, and 176 proteins quantified with two or more unique peptides were compared. The levels of most components of the proteome were similar in plasma and serum, and were stable under the storage conditions examined. However, it is necessary to consider the possibility of coagulation, as this affects the levels of platelet and hemolysis-related proteins. Interestingly, the blood proteome appears to be more stable than the blood metabolome, based on previously reported metabolomics data with same samples. These data will be helpful in designing protocols for blood sampling and for blood biomarker discovery and validation.

中文翻译：

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分析前处理中的差异对用SWATH-MS测定的血液蛋白质谱的影响。

本研究的目的是通过SWATH-MS分析评估人血清和血浆蛋白质组之间的差异，以及采血后在不同储存条件下人血浆蛋白的稳定性。当我们比较采血后立即准备的血浆和血清时，在176种定量蛋白质中，有95.5％的差异不到1.5倍。当我们比较通过血液在室温下储存0、15或30分钟，或在0-5°C下冷藏1、4或8小时后通过离心制备的血浆样品时，没有蛋白质显示出显着变化（q <0.05 ）除血红蛋白外，总计为1.5倍或更高。这些蛋白质在8小时内的单个样品中大量增加，可能是由于溶血所致。来自相同样品的数据比较表明，血液蛋白质组比血液代谢组更稳定。目前的结果表明，蛋白质组的大多数成分在血浆和血清中基本相同，并且在本研究中考察的储存条件下稳定。但是，重要的是要注意凝血程度以及血小板和溶血相关蛋白的水平。意义：采血后的分析前处理和储存条件预计会影响血液蛋白质组。因此，我们研究了人血清和血浆中蛋白质组的差异，以及人血浆蛋白在不同存储条件下的稳定性：在室温下0-30分钟，或在0-5°C下1-8小时，这可能反映了采血的临床情况。用SWATH-MS进行的蛋白质组学分析鉴定了342种蛋白质，并比较了用两种或两种以上独特肽定量的176种蛋白质。蛋白质组大多数成分的水平在血浆和血清中相似，并且在所研究的储存条件下稳定。但是，有必要考虑凝血的可能性，因为这会影响血小板和溶血相关蛋白的水平。有趣的是，根据先前报道的具有相同样品的代谢组学数据，血液蛋白质组似乎比血液代谢组更稳定。这些数据将有助于设计用于血液采样以及血液生物标志物发现和验证的方案。蛋白质组大多数成分的水平在血浆和血清中相似，并且在所研究的储存条件下稳定。但是，有必要考虑凝血的可能性，因为这会影响血小板和溶血相关蛋白的水平。有趣的是，根据先前报道的具有相同样品的代谢组学数据，血液蛋白质组似乎比血液代谢组更稳定。这些数据将有助于设计用于血液采样以及血液生物标志物发现和验证的方案。蛋白质组大多数成分的水平在血浆和血清中相似，并且在所考察的储存条件下稳定。但是，有必要考虑凝血的可能性，因为这会影响血小板和溶血相关蛋白的水平。有趣的是，根据先前报道的具有相同样品的代谢组学数据，血液蛋白质组似乎比血液代谢组更稳定。这些数据将有助于设计用于血液采样以及血液生物标志物发现和验证的方案。根据先前报道的相同样品的代谢组学数据，血液蛋白质组似乎比血液代谢组更稳定。这些数据将有助于设计用于血液采样以及血液生物标志物发现和验证的方案。根据先前报道的相同样品的代谢组学数据，血液蛋白质组似乎比血液代谢组更稳定。这些数据将有助于设计用于血液采样以及血液生物标志物发现和验证的方案。