Martinković Franjo, Popović Marin, Smolec Ozren, Mrljak Vladimir, Eckersall Peter David, Horvatić Anita
Faculty of Veterinary Medicine, University of Zagreb, Heinzelova 55, HR-10000 Zagreb, Croatia.
Department of Safety and Protection, Karlovac University of Applied Sciences, Trg Josipa Juraja Strossmayera 9, HR-47000 Karlovac, Croatia.
Metabolites. 2023 Feb 28;13(3):365. doi: 10.3390/metabo13030365.
Comprehensive profiling of serum proteome provides valuable clues of health status and pathophysiological processes, making it the main strategy in biomarker discovery. However, the high dynamic range significantly decreases the number of detectable proteins, obstructing the insights into the underlying biological processes. To circumvent various serum enrichment methods, obtain high-quality proteome wide information using the next-generation proteomic, and study host response in canine leishmaniosis, we applied data-independent acquisition mass spectrometry (DIA-MS) for deep proteomic profiling of clinical samples. The non-depleted serum samples of healthy and naturally -infected dogs were analyzed using the label-free 60-min gradient sequential window acquisition of all theoretical mass spectra (SWATH-MS) method. As a result, we identified 554 proteins, 140 of which differed significantly in abundance. Those were included in lipid metabolism, hematological abnormalities, immune response, and oxidative stress, providing valuable information about the complex molecular basis of the clinical and pathological landscape in canine leishmaniosis. Our results show that DIA-MS is a method of choice for understanding complex pathophysiological processes in serum and serum biomarker development.
血清蛋白质组的全面分析为健康状况和病理生理过程提供了有价值的线索,使其成为生物标志物发现的主要策略。然而,高动态范围显著减少了可检测蛋白质的数量,阻碍了对潜在生物学过程的深入了解。为了规避各种血清富集方法,使用新一代蛋白质组学获得高质量的全蛋白质组信息,并研究犬利什曼病中的宿主反应,我们应用数据非依赖采集质谱法(DIA-MS)对临床样本进行深度蛋白质组分析。使用无标记的60分钟梯度全理论质谱连续窗口采集(SWATH-MS)方法分析健康犬和自然感染犬的非耗尽血清样本。结果,我们鉴定出554种蛋白质,其中140种在丰度上有显著差异。这些蛋白质涉及脂质代谢、血液学异常、免疫反应和氧化应激,为犬利什曼病临床和病理情况的复杂分子基础提供了有价值的信息。我们的结果表明,DIA-MS是了解血清中复杂病理生理过程和血清生物标志物开发的首选方法。