A disposable bio-nano-chip using agarose beads for high performance immunoassays.
A disposable bio-nano-chip using agarose beads for high performance immunoassays.
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DOI：
10.1016/j.bios.2011.07.027
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发表时间：
2011-10-15
影响因子：
12.6
通讯作者：
McDevitt JT
中科院分区：
文献类型：
--
作者：
Du N;Chou J;Kulla E;Floriano PN;Christodoulides N;McDevitt JT
本文报道了一次性生物纳米芯片（BNC）的制作，一种由聚二甲基硅氧烷（PDMS）和基于硫醇的光学环氧树脂组成的微流控装置，既具有成本效益，又适用于高性能免疫分析。一种新的室温（RT）键合技术，利用，以实现PDMS和硫醇基环氧树脂层之间的不可逆共价键合，而在同一时间是兼容的插入琼脂糖珠传感器，选择性地排列在一个阵列中的金字塔微腔复制的硫醇薄膜层。在密封装置中，珠支撑环氧树脂膜夹在两个PDMS层之间，包括流体注入和排出通道。该装置中使用的琼脂糖珠传感器用抗C反应蛋白（CRP）抗体敏化，并进行荧光免疫分析以表征该装置的性能。根据器械规格使用计算流体动力学（CFD）对微珠穿透进行建模。实验数据显示，分析物渗透到100 μm的免疫复合物到280 μm直径的琼脂糖珠，这与模拟相关性良好。获得了剂量-响应曲线，并确定了测定的线性动态范围为1 ng/mL至50 ng/mL，检测限小于1 ng/mL。
This article reports on the fabrication of a disposable bio-nano-chip (BNC), a microfluidic device composed of polydimethylsiloxane (PDMS) and thiolene-based optical epoxy which is both cost-effective and suitable for high performance immunoassays. A novel room temperature (RT) bonding technique was utilized so as to achieve irreversible covalent bonding between PDMS and thiolene-based epoxy layers, while at the same time being compatible with the insertion of agarose bead sensors, selectively arranged in an array of pyramidal microcavities replicated in the thiolene thin film layer. In the sealed device, the bead-supporting epoxy film is sandwiched between two PDMS layers comprising of fluidic injection and drain channels. The agarose bead sensors used in the device are sensitized with anti-C-reactive protein (CRP) antibody, and a fluorescent sandwich-type immunoassay was run to characterize the performance of this device. Computational fluid dynamics (CFD) was used based on the device specifications to model the bead penetration. Experimental data revealed analyte penetration of the immunocomplex to 100 μm into the 280 μm diameter agarose beads, which correlated well with the simulation. A dose–response curve was obtained and the linear dynamic range of the assay was established over 1 ng/mL to 50 ng/mL with a limit of detection less than 1 ng/mL.
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