Total α-synuclein levels in human blood cells, CSF, and saliva determined by a lipid-ELISA
Total α-synuclein levels in human blood cells, CSF, and saliva determined by a lipid-ELISA
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DOI：
10.1007/s00216-016-9863-7
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发表时间：
2016-11-01
影响因子：
4.3
通讯作者：
Sharon, Ronit
中科院分区：
文献类型：
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作者：
Abd-Elhadi, Suaad;Basora, Misericordia;Sharon, Ronit
α-突触核蛋白 (α-Syn) 作为帕金森病 (PD) 生物标志物的有效性仍在研究中。用于捕获和定量人类样品中 α-Syn 蛋白的常规方法主要基于抗 α-Syn 抗体。针对 α-Syn 产生了特异性和有效的抗体。然而，抗α-Syn抗体的捕获可能仅限于特定表位识别，这归因于蛋白质结构或翻译后修饰。因此，基于抗体的 α-Syn 捕获方法引起了人们对其检测细胞内未折叠 α-Syn 池的功效的担忧。另一种方法是通过膜脂捕获α-Syn，即利用α-Syn的生化特性特异性结合膜脂并在结合后获得特征结构。我们使用固定化脂质捕获 α-Syn 来测定人类样本中的 α-Syn 水平。用于 α-Syn 捕获的脂质由磷脂酰肌醇 (PI)、磷脂酰丝氨酸 (PS) 和磷脂酰乙醇胺 (PE) 组成。在固定脂质中添加单唾液酸神经节苷脂（GM1 神经节苷脂）可显着改善 α-Syn 检测。捕获后，使用抗 α-Syn 抗体检测脂质结合的 α-Syn。通过脂质 ELISA 方法测定全血细胞 (WBC)、脑脊液 (CSF) 和唾液中的总 α-Syn 水平。
The validity of alpha-synuclein (alpha-Syn) as a biomarker for Parkinson's disease (PD) is still under investigation. Conventional methods for capture and quantitation of alpha-Syn protein in human samples are primarily based on anti-alpha-Syn antibodies. Specific and competent antibodies were raised against alpha-Syn. However, capture by anti-alpha-Syn antibodies may be limited to specific epitope recognition, attributed to protein structure or post-translational modifications. Hence, antibody-based methods for alpha-Syn capture raise a concern regarding their efficacy to detect the intracellular, unfolded alpha-Syn pool. An alternative is alpha-Syn capture by membrane lipids, i.e., to utilize the biochemical property of alpha-Syn to specifically bind membrane lipids and acquire a characteristic structure following binding. We determined alpha-Syn levels in human samples using immobilized lipids for alpha-Syn capture. The lipids used for alpha-Syn capture consist of phosphatidyl inositol (PI), phosphatidyl serine (PS), and phosphatidyl ethanolamine (PE). Addition of mono-sialoganglioside, GM1 ganglioside, to the immobilized lipids significantly improved alpha-Syn detection. Following capture, the lipid-bound alpha-Syn was detected using an anti-alpha-Syn antibody. Total alpha-Syn levels in whole blood cells (WBC), cerebrospinal fluid (CSF), and saliva were determined by the lipid-ELISA method.