Enzyme-linked immunosorbent assay-based method to quantify the association of small molecules with aggregated amyloid peptides.
Enzyme-linked immunosorbent assay-based method to quantify the association of small molecules with aggregated amyloid peptides.
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DOI：
10.1021/ac2030859
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发表时间：
2012-02-07
影响因子：
7.4
通讯作者：
Yang, Jerry
中科院分区：
文献类型：
--
作者：
Capule, Christina C.;Yang, Jerry
本文描述了一种用于定量小分子与聚集的β淀粉样蛋白（Aβ）肽结合的简单ELISA方案。淀粉样蛋白靶向小分子作为治疗或诊断神经退行性疾病如阿尔茨海默病的潜在药物引起了广泛的兴趣。然而，由于缺乏评价小分子-淀粉样蛋白结合相互作用的通用方法，迄今为止已经研究的淀粉样蛋白靶向分子的数量受到了极大的限制。在这里，我们展示了一个通用的方法来量化小分子-淀粉样蛋白结合的相互作用，通过修改后的定量ELISA协议。该方案的一个关键特征是用空气等离子体处理商业ELISA板，以帮助在将这些肽固定到聚苯乙烯表面上时维持聚集的Aβ的所需β-折叠含量。我们在这些空气等离子体处理的平板上开发了基于ELISA的竞争试验，并评估了五种先前已知的淀粉样蛋白结合小分子与聚集的Aβ的结合。我们表明，这种一般的ELISA为基础的竞争测定可用于量化小分子-淀粉样蛋白结合的相互作用，在低纳摩尔低微摩尔范围内，这是目前开发的许多淀粉样蛋白靶向诊断剂的亲和力的典型范围。这种用于定量小分子与聚集Aβ肽相互作用的简单方案克服了先前报道的光谱或放射性测定的许多局限性，因此可能有助于筛选和评价比先前可能的结构更多样化的淀粉样蛋白靶向剂。
This paper describes a simple ELISA protocol for quantifying the binding of small molecules to aggregated Amyloid-β (Aβ) peptides. Amyloid-targeting small molecules have attracted wide interest as potential agents for the treatment or diagnosis of neurodegenerative disorders such as Alzheimer’s disease. The lack of general methods to evaluate small molecule-amyloid binding interactions, however, has significantly limited the number of amyloid-targeting molecules that have been studied to date. Here, we demonstrate a general method to quantify small molecule-amyloid binding interactions via a modified quantitative ELISA protocol. A key feature of this protocol is the treatment of commercial ELISA plates with an air plasma to help maintain the desired β-sheet content of the aggregated Aβ upon immobilization of these peptides on to the polystyrene surface. We developed an ELISA-based competition assay on these air plasma-treated plates and evaluated the binding of five previously known amyloid-binding small molecules to aggregated Aβ. We show that this general ELISA-based competition assay can be used to quantify small molecule-amyloid binding interactions in the low nanomolar to low micromolar range, which is the typical range of affinities for many amyloid-targeting diagnostic agents under current development. This simple protocol for quantifying the interaction of small molecules with aggregated Aβ peptides overcomes many limitations of previously reported spectroscopic or radioactivity assays, and may, therefore, facilitate the screening and evaluation of a more structurally diverse set of amyloid-targeting agents than had previously been possible.
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影响因子：
4.6
作者：
Giacomelli, CE;Norde, W
通讯作者：
Norde, W
影响因子：
3.6
作者：
HOLLANDER, Z;KATCHALSKIKATZIR, E
通讯作者：
KATCHALSKIKATZIR, E
DOI：
10.1001/jama.2010.2008
发表时间：
2011-01-19
期刊：
JAMA
影响因子：
--
作者：
Clark CM;Schneider JA;Bedell BJ;Beach TG;Bilker WB;Mintun MA;Pontecorvo MJ;Hefti F;Carpenter AP;Flitter ML;Krautkramer MJ;Kung HF;Coleman RE;Doraiswamy PM;Fleisher AS;Sabbagh MN;Sadowsky CH;Reiman EP;Zehntner SP;Skovronsky DM;AV45-A07 Study Group
通讯作者：
AV45-A07 Study Group
DOI：
10.1073/pnas.96.7.3688
发表时间：
1999-03-30
影响因子：
11.1
作者：
Kowalewski, T;Holtzman, DM
通讯作者：
Holtzman, DM
影响因子：
15
作者：
Childers, W. Seth;Mehta, Anil K.;Lynn, David G.
通讯作者：
Lynn, David G.