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从血清中高效提取完整的 HSA-Aβ 肽复合物:寻找白蛋白组学生物标志物。

Efficient extraction of intact HSA-Aβ peptide complexes from sera: Toward albuminome biomarker identification.

机构信息

Université Paris-Saclay, CNRS, Institut Galien Paris Sud, 92296, Châtenay-Malabry, France.

Université Paris-Saclay, CNRS, Institut Galien Paris Sud, 92296, Châtenay-Malabry, France.

出版信息

Talanta. 2020 Aug 15;216:121002. doi: 10.1016/j.talanta.2020.121002. Epub 2020 Apr 9.

Abstract

The performances of three commercial albumin extraction methods for the isolation of intact albumin-amyloid beta peptide (HSA-Aβ) complexes from serum were compared using different analytical approaches. To determine the extraction yield, the repeatability and the selectivity of the extraction procedures, a capillary electrophoresis coupled to UV detection method was developed. For the evaluation of the specificity and integrity of the extracted HSA-Aβ complexes, SDS-PAGE, hybrid and ultra-sensitive ELISA experiments were conducted. All the extraction methods showed different characteristics depending on their chemical binding affinities toward albumin. The ProteoExtract Albumin Depletion kit extracted albumin with a high repeatability but was not efficient for the extraction of intact HSA-Aβ complexes. The PureProteome Albumin magnetic beads showed a high specificity toward HSA thanks to the grafting of anti-HSA antibodies on their surface but tended to dissociate HSA from Aβ peptides. The Pierce Albumin depletion kit showed a high extraction yield, no selectivity towards the different albumin proteoforms and proved to be the most efficient method for the extraction of intact HSA-Aβ complexes from serum.

摘要

比较了三种商业白蛋白提取方法在血清中分离完整的白蛋白-淀粉样β肽(HSA-Aβ)复合物的性能,使用了不同的分析方法。为了确定提取产率、提取程序的重复性和选择性,开发了一种毛细管电泳与紫外检测相结合的方法。为了评估提取的 HSA-Aβ 复合物的特异性和完整性,进行了 SDS-PAGE、杂交和超灵敏 ELISA 实验。所有的提取方法都因其对白蛋白的化学结合亲和力而表现出不同的特性。ProteoExtract 白蛋白去除试剂盒对白蛋白的提取具有很高的重复性,但对完整 HSA-Aβ 复合物的提取效率不高。PureProteome 白蛋白磁珠由于其表面接枝了抗 HSA 抗体,对 HSA 具有很高的特异性,但往往会使 HSA 从 Aβ 肽中解离。Pierce 白蛋白去除试剂盒显示出较高的提取产率,对不同的白蛋白蛋白形式没有选择性,并且被证明是从血清中提取完整 HSA-Aβ 复合物的最有效方法。

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