Konstantinos Kalogeropoulos, Simonas Savickas, Aleksander M Haack, Cathrine A Larsen, Jacek Mikosiński, Erwin M Schoof, Hans Smola, Louise Bundgaard, Ulrich Auf dem Keller
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引用次数: 0
Abstract
Targeted proteomics methods have been greatly improved and refined over the last decade and are becoming increasingly the method of choice in protein and peptide quantitative assays. Despite the tremendous progress, targeted proteomics assays still suffer from inadequate sensitivity for lower abundant proteins and throughput, especially in complex biological samples. These attributes are essential for establishing targeted proteomics methods at the forefront of clinical use. Here, we report an assay utilizing the SureQuantTM internal standard triggered targeted method on a latest generation mass spectrometer coupled with an EvoSep One liquid chromatography platform, which displays high sensitivity and a high throughput of 100 samples per day (SPD). We demonstrate the robustness of this method by quantifying proteins spanning six orders of magnitude in human wound fluid exudates, a biological fluid that exhibits sample complexity and composition similar to plasma. Among the targets quantified were low-abundance proteins such at tumor necrosis factor A (TNFA) and interleukin 1-β (IL1B), highlighting the value of this method in the quantification of trace amounts of invaluable biomarkers that were until recently hardly accessible by targeted proteomics methods. Taken together, this method extends the toolkit of targeted proteomics assays and will help to drive forward mass spectrometry-based proteomics biomarker quantification.
利用快速液相色谱 SureQuant™ IS 靶向定量技术对体液中的生物标记物进行高通量、高灵敏度监测。
在过去的十年中,靶向蛋白质组学方法得到了极大的改进和完善,并逐渐成为蛋白质和肽定量检测的首选方法。尽管取得了巨大进步,但靶向蛋白质组学检测仍存在对低丰度蛋白质的灵敏度和通量不足的问题,尤其是在复杂的生物样本中。这些特性对于将靶向蛋白质组学方法应用于临床至关重要。在此,我们报告了一种利用 SureQuantTM 内标触发靶向方法在最新一代质谱仪和 EvoSep One 液相色谱平台上进行的检测,该方法具有高灵敏度和每天 100 个样品 (SPD) 的高通量。我们通过量化人体伤口渗出液(一种样本复杂、成分类似血浆的生物液体)中跨越六个数量级的蛋白质,证明了这种方法的稳健性。定量的目标蛋白包括肿瘤坏死因子 A (TNFA) 和白细胞介素 1-β (IL1B)等低丰度蛋白,突出了这种方法在定量痕量宝贵生物标志物方面的价值,直到最近,靶向蛋白质组学方法还很难获得这些生物标志物。总之,这种方法扩展了靶向蛋白质组学测定的工具包,将有助于推动基于质谱的蛋白质组学生物标记物定量。
期刊介绍:
The mission of MCP is to foster the development and applications of proteomics in both basic and translational research. MCP will publish manuscripts that report significant new biological or clinical discoveries underpinned by proteomic observations across all kingdoms of life. Manuscripts must define the biological roles played by the proteins investigated or their mechanisms of action.
The journal also emphasizes articles that describe innovative new computational methods and technological advancements that will enable future discoveries. Manuscripts describing such approaches do not have to include a solution to a biological problem, but must demonstrate that the technology works as described, is reproducible and is appropriate to uncover yet unknown protein/proteome function or properties using relevant model systems or publicly available data.
Scope:
-Fundamental studies in biology, including integrative "omics" studies, that provide mechanistic insights
-Novel experimental and computational technologies
-Proteogenomic data integration and analysis that enable greater understanding of physiology and disease processes
-Pathway and network analyses of signaling that focus on the roles of post-translational modifications
-Studies of proteome dynamics and quality controls, and their roles in disease
-Studies of evolutionary processes effecting proteome dynamics, quality and regulation
-Chemical proteomics, including mechanisms of drug action
-Proteomics of the immune system and antigen presentation/recognition
-Microbiome proteomics, host-microbe and host-pathogen interactions, and their roles in health and disease
-Clinical and translational studies of human diseases
-Metabolomics to understand functional connections between genes, proteins and phenotypes