Combined top-down and bottom-up mass spectrometric approach to characterization of biomarkers for renal disease
Combined top-down and bottom-up mass spectrometric approach to characterization of biomarkers for renal disease
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DOI：
10.1021/ac050983o
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发表时间：
2005-11-15
影响因子：
7.4
通讯作者：
Marshall, AG
中科院分区：
文献类型：
--
作者：
Chalmers, MJ;Mackay, CL;Marshall, AG
在这里，我们描述了一种基于自上而下和自下而上分析的生物标志物发现和结构表征的质谱学(MS)方法。毛细管电泳联用飞行时间质谱仪(TOF-MS)对人体尿液中数千种多肽进行了分离和质量检测。对健康对照样本和局灶性节段性肾小球硬化、膜性肾小球肾炎、微小病变、IgA肾病和糖尿病肾病患者之间的差异进行统计分析，证实了对照和每种疾病的多个生物标志物。为了识别这些生物标志物，我们采用了制备性CE，实现了直接输注ESI MS分析，然后在必要时进行样品处理和重新分析。我们展示了串联傅里叶变换离子回旋共振(FT-ICR)MS如何识别这些有时很大(>；8 kDa)的生物标记物。至关重要的是，我们保持了CE TOF MS数据和用于生物标记物识别的ICR数据之间的连接。
Here we describe a mass spectrometry (MS) approach for biomarker discovery and structural characterization, based on both top-down and bottom-up analyses. Capillary electrophoresis (CE) coupled to electrospray ionization (ESI) time-of-flight (TOF) MS serves to separate and mass-measure the thousands of polypeptides contained in human urine. Statistical analysis of the differences between healthy control samples and patients with focal-segmental glomerulosclerosis, membranous glomerulonephritis, minimal change disease, IgA nephropathy, and diabetic nephropathy validates multiple biomarkers for the control and each of the diseases. To identify those biomarkers, we employ preparative CE, enabling direct infusion ESI MS analysis, followed by sample manipulation and reanalysis where necessary. We show how tandem Fourier transform ion cyclotron resonance (FT-ICR) MS identifies these sometimes large (> 8 kDa) biomarkers. Critically, we maintain connectivity between the CE TOF MS data and the ICR data used for biomarker identification.