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Comparison of proteins in whole blood and dried blood spot samples by LC/MS/MS

机译:通过LC / MS / MS比较全血和干血斑样品中的蛋白质

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摘要

Dried blood spot (DBS) sampling methods are desirable for population-wide biomarker screening programs because of their ease of collection, transportation, and storage. Immunoassays are traditionally used to quantify endogenous proteins in these samples but require a separate assay for each protein. Recently, targeted mass spectrometry (MS) has been proposed for generating highly-multiplexed assays for biomarker proteins in DBS samples. In this work, we report the first comparison of proteins in whole blood and DBS samples using an untargeted MS approach. The average number of proteins identified in undepleted whole blood and DBS samples by liquid chromatography (LC)/MS/MS was 223 and 253, respectively. Protein identification repeatability was between 77 %-92 % within replicates and the majority of these repeated proteins (70 %) were observed in both sample formats. Proteins exclusively identified in the liquid or dried fluid spot format were unbiased based on their molecular weight, isoelectric point, aliphatic index, and grand average hydrophobicity. In addition, we extended this comparison to include proteins in matching plasma and serum samples with their dried fluid spot equivalents, dried plasma spot (DPS), and dried serum spot (DSS). This work begins to define the accessibility of endogenous proteins in dried fluid spot samples for analysis by MS and is useful in evaluating the scope of this new approach. [Figure not available: see fulltext.]
机译:干血斑(DBS)采样方法因其易于收集,运输和存储而对于整个人群的生物标志物筛选程序而言是理想的。免疫测定传统上用于定量这些样品中的内源性蛋白质,但需要对每种蛋白质进行单独的测定。最近,已经提出了靶向质谱分析法(MS)来生成DBS样品中生物标志物蛋白的高度多重测定。在这项工作中,我们报告了使用非靶向MS方法对全血和DBS样品中蛋白质的首次比较。通过液相色谱(LC)/ MS / MS在未消耗的全血和DBS样品中鉴定出的平均蛋白质数量分别为223和253。重复中的蛋白质鉴定重复性在77%-92%之间,并且在两种样品形式中均观察到大多数这些重复蛋白质(70%)。基于分子量,等电点,脂肪族指数和平均疏水性,以液体或干燥液体斑点形式专门鉴定的蛋白质没有偏见。此外,我们将此比较扩展到包括匹配血浆和血清样品中的蛋白质及其干液斑点当量,干血浆斑点(DPS)和干血清斑点(DSS)。这项工作开始确定干燥液体斑点样品中内源蛋白的可及性,以供MS分析,并有助于评估这种新方法的范围。 [图不可用:请参见全文。]

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