A selected reaction monitoring (SRM)-based method for absolute quantification of Aβ38, Aβ40, and Aβ42 in cerebrospinal fluid of Alzheimer's disease patients and healthy controls.
A selected reaction monitoring (SRM)-based method for absolute quantification of Aβ38, Aβ40, and Aβ42 in cerebrospinal fluid of Alzheimer's disease patients and healthy controls.
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基于选择反应监测 (SRM) 的方法，用于对阿尔茨海默病患者和健康对照者脑脊液中的 Aβ38、Aβ40 和 Aβ42 进行绝对定量。
DOI：
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发表时间：
2013
期刊：
影响因子：
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通讯作者：
J. Gobom
中科院分区：
文献类型：
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作者：
Josef Pannee;E. Portelius;M. Oppermann;Alan R. Atkins;M. Hornshaw;I. Zegers;P. Höjrup;L. Minthon;O. Hansson;H. Zetterberg;K. Blennow;J. Gobom
阿尔茨海默病（AD）的脑脊液（CSF）生物标志物越来越多地用于研究中心，临床试验和临床环境。然而，由于缺乏跨分析平台的标准化以及淀粉样蛋白（Aβ）与基质蛋白结合以及自聚集的干扰，它们的广泛使用受到阻碍。在本文中，我们报告了一种抗基质效应方法，用于测量人CSF中AD相关的42种氨基酸Aβ（Aβ42）以及Aβ40和Aβ38，该方法基于使用选择反应监测（SRM）的质谱定量。通过固相萃取制备样品，并使用稳定同位素标记的Aβ肽作为内标进行定量。该方法的诊断性能进行了评估两个独立的临床材料与研究志愿者谁是认知正常和AD患者轻度至中度痴呆。该方法的分析特征包括Aβ42的定量下限为62.5 pg/mL，变异系数低于10%。在一项针对AD患者和对照的初步研究中，我们证实了与Aβ42水平降低的疾病相关性，与ELISA获得的结果相似，使用Aβ42/Aβ40比值获得了更好的分离。开发的检测方法具有灵敏度，不受基质效应的影响，能够绝对定量CSF中的Aβ42、Aβ40和Aβ38，同时保留区分AD患者和对照的能力。我们建议这种基于SRM的人CSF中Aβ肽定量方法对临床研究和试验有价值。
Cerebrospinal fluid (CSF) biomarkers for Alzheimer's disease (AD) are increasingly used in research centers, clinical trials, and clinical settings. However, their broad-scale use is hampered by lack of standardization across analytical platforms and by interference from binding of amyloid-β (Aβ) to matrix proteins as well as self-aggregation. Here, we report on a matrix effect-resistant method for the measurement of the AD-associated 42 amino acid species of Aβ (Aβ42), together with Aβ40 and Aβ38 in human CSF based on mass spectrometric quantification using selected reaction monitoring (SRM). Samples were prepared by solid-phase extraction and quantification was performed using stable-isotope labeled Aβ peptides as internal standards. The diagnostic performance of the method was evaluated on two independent clinical materials with research volunteers who were cognitively normal and AD patients with mild to moderate dementia. Analytical characteristics of the method include a lower limit of quantification of 62.5 pg/mL for Aβ42 and coefficients of variations below 10%. In a pilot study on AD patients and controls, we verified disease-association with decreased levels of Aβ42 similar to that obtained by ELISA and even better separation was obtained using the Aβ42/Aβ40 ratio. The developed assay is sensitive and is not influenced by matrix effects, enabling absolute quantification of Aβ42, Aβ40, and Aβ38 in CSF, while it retains the ability to distinguish AD patients from controls. We suggest this SRM-based method for Aβ peptide quantification in human CSF valuable for clinical research and trials.
影响因子：
4.4
作者：
Pan S;Aebersold R;Chen R;Rush J;Goodlett DR;McIntosh MW;Zhang J;Brentnall TA
通讯作者：
Brentnall TA