Capillary-driven multiparametric microfluidic chips for one-step immunoassays
Capillary-driven multiparametric microfluidic chips for one-step immunoassays
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DOI：
10.1016/j.bios.2011.06.016
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发表时间：
2011-09-15
影响因子：
12.6
通讯作者：
Delamarche, Emmanuel
中科院分区：
文献类型：
--
作者：
Gervais, Luc;Hitzbleck, Martina;Delamarche, Emmanuel
在这里，我们提出了一种用于“一步”免疫测定的毛细血管驱动微流体芯片。该芯片允许轻松修改几个测定参数，如样品的流速，用于测试的样品的体积，以及用于检测分析物的试剂和受体的类型。因此，我们将这样的芯片称为多参数芯片，并通过抗C-反应蛋白（CRP）检测抗体（dAb）的整合和释放以及含有CRP的样品跨抗CRP捕获抗体（cAb）的线的分流来说明这一概念。微流控芯片在Si中制造，并用cAb图案化的聚二甲基硅氧烷（PDMS）密封。该微流控芯片尺寸为1.7 × 3.4 cm（2），能够在6个平行流路中分析20 μ L人血清，流速范围为3.3 nL·s（-1）~ 0.46 nL·s（-1）。使用喷墨点样器将10.6 nL的dAb溶液存款沉积在芯片主流路附近的结构中。随后在人血清流中dAb的不对称释放通过具有9个混合回路和2.8 mm x 0.78 mm的占地面积的Dean流动混合器来补偿。在靠近点样区域的流动路径的一半中存在的dAb的量从混合器入口处的83%降低到混合器之后的区域中的52%。然后将样品平均分配到6个反应室中，并通过连接通道进入2 μ L毛细管泵。连接通道的液压阻力被设计为改变流速，因此CRP-dAb复合物的捕获动力学，从10分钟到72分钟。增加的孵育时间导致反应室中的检测信号增加四倍，孵育时间更长。这里提出的概念是灵活的，适用于实现各种表面荧光免疫分析的毛细管驱动的微流控芯片。(C)2011 Elsevier B. V.保留所有权利。
Here we present a capillary-driven microfluidic chip for "one-step" immunoassays. The chip allows for easy modification of several assay parameters such as the flow rates of sample, the volumes of samples for tests, and the type of reagents and receptors for detecting analytes. We therefore term such a chip a multiparametric chip and illustrate this concept with the integration and release of anti-C-reactive protein (CRP) detection antibodies (dAbs) together with splitting flow of samples containing CRP across lines of anti-CRP capture antibodies (cAbs). The microfluidic chip is fabricated in Si and is sealed with polydimethylsiloxane (PDMS) patterned with cAbs. The microfluidic chip is similar to 1.7 x 3.4 cm(2) and is capable of analyzing 20 mu L of human serum in 6 parallel flow paths with a range of flow rates from 3.3 nL s(-1) to 0.46 nL s(-1). An inkjet spotter was used to deposit 10.6 nL of dAb solution in a structure vicinal to the main flow path of the chip. The consequent asymmetric release of dAbs in a stream of human serum is compensated by a Dean flow mixer having 9 mixing loops and a footprint of 2.8 mm x 0.78 mm. The quantity of dAb present in the half of the flow path close to the spotting region decreases from 83% at the entrance of the mixer to 52% in the region after the mixer. The sample is then equally split into 6 reaction chambers and proceeds via connecting channels to 2 mu L capillary pumps. The hydraulic resistance of the connecting channels is designed to vary flow rates, and therefore the kinetics of capture of CRP-dAb complexes, from 10 min to 72 min. The increased incubation time leads to a fourfold increase in detection signal in the reaction chamber with the longer incubation time. The concept presented here is flexible and suited for implementing various surface fluorescence immunoassays on a capillary-driven microfluidic chip. (C) 2011 Elsevier B.V. All rights reserved.