Multiplexed Assay for Proteins Based on Sequestration Electrochemistry Using the Protein Binding Electroactive Magnetic Microbeads-猫眼课题宝

DOI：

10.2116/analsci.28.77

发表时间：

2012-01

期刊：

Analytical Sciences

影响因子：

1.6

通讯作者：

中科院分区：

化学4区

文献类型：

作者：

H. Kuramitz;Yoko Mawatari;Mariwo Ikeuchi;O. Kutomi;N. Hata;S. Taguchi;K. Sugawara

采用顺磁性微珠电化学结合法同时检测两种蛋白质。该分析的原理基于螯合电化学。制备了与电活性化合物和配体偶联的蛋白结合电活性磁性微球，以与靶蛋白特异性结合。选择亲和素-生物素和大豆凝集素（SBA）-半乳糖胺作为模型蛋白-配体系统。将生物素/硫代氨基酚和半乳糖胺/二茂铁修饰在顺磁性微球上，构建亲和素结合电活性磁性微球（ABEMMb）和SBA结合电活性磁性微球（SBEMMb）。用Nd-Fe-B磁结合碳糊旋转圆盘电极测量了这些功能化微球的伏安响应。使用旋转盘电极系统在微升液滴中进行测量。通过ABEMMb和SBEMMb与靶蛋白结合引起的电流响应降低，同时检测亲和素和SBA。抗生物素蛋白和SBA的检测限分别为4 × 10^-10和2 × 10^-10 M。

The paramagnetic microbead-based electrochemical binding assay was demonstrated for detecting two kinds of protein simultaneously. The principle of this assay is based on the sequestration electrochemistry. The protein binding electroactive magnetic microbeads which are conjugated with an electroactive compound and a ligand to bind specifically with a target protein were prepared. The avidin-biotin and soybean agglutinin (SBA)-galactosamine were chosen as model protein-ligand systems. The avidin binding electroactive magnetic microbead (ABEMMb) and SBA binding electroactive magnetic microbead (SBEMMb) are constructed by biotin/thionine and galactosamine/ferrocene modified on paramagnetic microbeads. The voltammetric response for these functionalized microbeads was measured by the Nd-Fe-B magnet-incorporating carbon paste rotating disk electrode. The measurements were performed in a microliter droplet using a rotating disk electrode system. Avidin and SBA were simultaneously detected by the decrease in the current responses from the reduction of ABEMMb and SBEMMb that was caused by the binding with target proteins. The limits of detection for avidin and SBA were 4 × 10^−10 and 2 × 10^−10 M, respectively.

Multiplexed Assay for Proteins Based on Sequestration Electrochemistry Using the Protein Binding Electroactive Magnetic Microbeads

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