Redox-magnetohydrodynamics, flat flow profile-guided enzyme assay detection: toward multiple, parallel analyses.
Redox-magnetohydrodynamics, flat flow profile-guided enzyme assay detection: toward multiple, parallel analyses.
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氧化还原磁流体动力学、平流剖面引导酶测定检测：走向多重并行分析。
DOI：
10.1021/ac502014t
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发表时间：
2014
影响因子：
7.4
通讯作者：
I. Fritsch
中科院分区：
文献类型：
--
作者：
V. Sahore;I. Fritsch
一个概念验证的超顺磁微珠-酶复合物与氧化还原磁流体动力学（MHD）泵送的微流体相结合，利用芯片下的磁体（0.56 T）和均匀的平坦流型，作为在芯片上开发多个并行化学分析的第一步，而不需要独立的通道。超顺磁磁珠用碱性磷酸酶（生化分析中常用的酶标记）衍生化，并在芯片上的三个不同位置进行磁固定，其中一个直接在通往检测器的路径上，另外两个位置与检测器直径相邻但远离路径，距离为检测器直径的5倍。在含有氧化还原物质[Ru(NH3)6]（3+/2+）和Tris缓冲液的溶液中，电活性对氨基酚由其电活性前体对氨基苯基磷酸在珠状酶复合物中酶促生成，并通过氧化还原- mhd运输，并在流动路径上位于珠状复合物下游2 mm的直径312 μm的金微盘上用方波伏安法检测。相反偏压的泵送电极由2.5 cm长的金带组成，间隔5.6 mm，位于含有球-酶复合物和检测位点的活性流动区域的两侧。来自相邻路径的信号仅为直接路径的20%，当泵送电极不活动时≤8%。
A proof-of-concept superparamagnetic microbead-enzyme complex was integrated with microfluidics pumped by redox-magneto-hydrodynamics (MHD) to take advantage of the magnet (0.56 T) beneath the chip and the uniform flat flow profile, as a first step toward developing multiple, parallel chemical analyses on a chip without the need for independent channels. The superparamagnetic beads were derivatized with alkaline phosphatase (a common enzyme label for biochemical assays) and magnetically immobilized at three different locations on the chip with one directly on the path to the detector and the other two locations adjacent to, but off the path, by a distance >5 times the detector diameter. Electroactive p-aminophenol, enzymatically generated at the bead-enzyme complex from its electroinactive precursor p-aminophenyl phosphate in a solution containing a redox species [Ru(NH3)6](3+/2+) for pumping and Tris buffer, was transported by redox-MHD and detected with square wave voltammetry at a 312 μm diameter gold microdisk stationed 2 mm downstream from the bead-complex on the flow path. Oppositely biased pumping electrodes, consisting of 2.5 cm long gold bands and separated by 5.6 mm, flanked the active flow region containing the bead-enzyme complex and detection site. The signal from adjacent paths was only 20% of that for the direct path and ≤8% when pumping electrodes were inactive.
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