Abstract：
The use of protein biomarkers in blood for clinical settings is limited by the cost and accessibility of traditional venipuncture sampling. The dried blood spot (DBS) technique offers a less invasive and more accessible alternative. However, protein stability in DBS has not been well evaluated. Herein, we deployed a quantitative LC-MS/MS system to construct proteomic atlases of whole blood, DBSs, plasma, and blood cells. Approximately 4% of detected proteins\' abundance was significantly altered during blood drying into blood spots, with overwhelming disturbances in cytoplasmic fraction. We also reported a novel finding suggesting a decrease in the level of membrane/cytoskeletal proteins (SLC4A1, RHAG, DSC1, DSP, and JUP) and an increase in the level of proteins (ATG3, SEC14L4, and NRBP1) related to intracellular trafficking. Furthermore, we identified 19 temporally dynamic proteins in DBS samples stored at room temperature for up to 6 months. There were three declined cytoskeleton-related proteins (RDX, SH3BGRL3, and MYH9) and four elevated proteins (XPO7, RAN, SLC2A1, and SLC29A1) involved in cytoplasmic transport as representatives. The instability was governed predominantly by hydrophilic proteins and enhanced significantly with an increasing storage time. Our analyses provide comprehensive knowledge of both short- and long-term storage stability of DBS proteins, forming the foundation for the widespread use of DBS in clinical proteomics and other analytical applications.
摘要：
在临床环境中使用血液中的蛋白质生物标志物受到传统静脉穿刺取样的成本和可及性的限制。干血斑（DBS）技术提供了一种侵入性较小且更容易获得的替代方法。然而,蛋白质在DBS中的稳定性尚未得到很好的评估。在这里，我们部署了定量LC-MS/MS系统来构建全血的蛋白质组学图谱，DBSs,等离子体,和血细胞。大约4%的检测到的蛋白质丰度在血液干燥成血斑期间发生了显著变化，细胞质部分有压倒性的干扰。我们还报道了一个新发现，表明膜/细胞骨架蛋白（SLC4A1，RHAG，DSC1,DSP,和JUP）以及与细胞内运输有关的蛋白质（ATG3，SEC14L4和NRBP1）水平的增加。此外，我们在室温保存长达6个月的DBS样品中鉴定出19种时间动态蛋白.有三种细胞骨架相关蛋白下降(RDX，SH3BGRL3和MYH9）和四种升高的蛋白质（XPO7，RAN，SLC2A1和SLC29A1）作为代表参与细胞质运输。不稳定性主要由亲水性蛋白质控制，并且随着储存时间的增加而显着增强。我们的分析提供了DBS蛋白的短期和长期储存稳定性的全面知识，为DBS在临床蛋白质组学和其他分析应用中的广泛应用奠定了基础。
