使用质谱参考测量程序对脑脊液中的Aβ1-42进行绝对定量
Absolute Quantification of Aβ1-42 in CSF Using a Mass Spectrometric Reference Measurement Procedure.
作者信息
Pannee Josef, Blennow Kaj, Zetterberg Henrik, Portelius Erik
机构信息
Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital; Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital.
Institute of Neuroscience and Physiology, Department of Psychiatry and Neurochemistry, The Sahlgrenska Academy at University of Gothenburg, Sahlgrenska University Hospital; Clinical Neurochemistry Laboratory, Sahlgrenska University Hospital; UCL Institute of Neurology, Queen Square;
出版信息
J Vis Exp. 2017 Mar 21(121):55386. doi: 10.3791/55386.
Alzheimer's disease (AD) is the most common neurodegenerative disease among the elderly and accounts for 60-80% of all cases of dementia. Currently, the diagnosis of AD is based on cognitive tests and mental state exams, but the peptide amyloid-beta (Aβ) in cerebrospinal fluid (CSF) is increasingly used in clinical trials and settings. As for most protein and peptide biomarkers, quantification is performed using antibody-based techniques, such as enzyme-linked immunosorbent assay (ELISA). However, intra- and inter-laboratory variability in these assays hamper its use as a diagnostic marker in clinical routine. An antibody-independent Reference Measurement Procedure (RMP) was developed based on solid-phase extraction (SPE) and liquid chromatography (LC)-tandem mass spectrometry (MS/MS), where stable, isotope-labeled Aβ peptides were used as internal standards, enabling absolute quantification. A high-resolution quadrupole-orbitrap hybrid instrument was used for the measurements. The method allows for the quantification of CSF Aβ1-42 between 150-4,000 pg/mL.
阿尔茨海默病（AD）是老年人中最常见的神经退行性疾病，占所有痴呆病例的60-80%。目前，AD的诊断基于认知测试和精神状态检查，但脑脊液（CSF）中的β淀粉样肽（Aβ）在临床试验和临床环境中越来越多地被使用。对于大多数蛋白质和肽生物标志物，定量是使用基于抗体的技术进行的，如酶联免疫吸附测定（ELISA）。然而，这些测定方法在实验室内和实验室间的变异性阻碍了其在临床常规中用作诊断标志物。基于固相萃取（SPE）和液相色谱（LC）-串联质谱（MS/MS）开发了一种独立于抗体的参考测量程序（RMP），其中使用稳定的、同位素标记的Aβ肽作为内标，实现绝对定量。使用高分辨率四极杆-轨道阱混合仪器进行测量。该方法可对150-4000 pg/mL的脑脊液Aβ1-42进行定量。
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