Laboratory of Microsystems, Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
Anal Chem. 2010 Dec 15;82(24):9989-97. doi: 10.1021/ac102149f. Epub 2010 Oct 22.
We propose a simple microfluidic device for protein preconcentration based on the electrokinetic trapping principle. It comprises a narrow Nafion strip that is simply cut from a commercial membrane and is integrated into a molded poly(dimethylsiloxane) (PDMS) microfluidic structure using a guiding channel. Mechanically clamping the PDMS/Nafion assembly with a glass substrate results in a rapid prototypable, leak-tight, and easily disposable device. Our device preconcentrates negatively charged fluorescent proteins located at the anodic microfluidic compartment side of the Nafion strip within a few minutes and up to a concentration factor of 10(4). Moreover, we present a numerical study of the preconcentration effect by solving the coupled Poisson, Nernst-Planck, and Navier-Stokes equations for our type of device, which provides microscopic insight into the mechanism of preconcentration. The electrical field across the ion-permselective Nafion generates concentration polarization, i.e., ion depletion at the anodic side and ion enrichment at the cathodic side for both types of ions, with a local excess of mobile positive ions in the depleted concentration polarization zone, inducing a nonequilibrium electrical double layer in close proximity to the Nafion membrane. A voltage difference applied over the anodic compartment is used to generate the electrophoretic flow velocity of the negatively charged tracer biomolecules. This, in combination with the electroosmotic flow in the opposite direction, which originates from the fixed charges on the channel walls and the induced space charge near the membrane, provides the basis for the local preconcentration of the negative tracer biomolecules.
我们提出了一种基于电动捕获原理的简单蛋白质预浓缩微流控装置。它由一个从商业膜上简单切割下来的窄 Nafion 条带组成,并通过导向通道集成到模制的聚二甲基硅氧烷 (PDMS) 微流控结构中。通过玻璃基底机械夹紧 PDMS/Nafion 组件,可得到快速原型、防漏且易于处理的设备。我们的设备可在几分钟内将带负电荷的荧光蛋白浓缩到 Nafion 条带的阳极微流控隔间侧,浓缩倍数高达 10(4)。此外,我们通过求解我们这种类型设备的耦合泊松、Nernst-Planck 和纳维-斯托克斯方程,对预浓缩效果进行了数值研究,为预浓缩机制提供了微观见解。离子选择性 Nafion 中的电场会产生浓度极化,即在阳极侧耗尽离子,在阴极侧富集离子,对于两种类型的离子,在耗尽的浓度极化区都存在局部过量的可移动正离子,在靠近 Nafion 膜的地方产生非平衡电双层。在阳极隔间上施加电压差,用于产生带负电荷的示踪生物分子的电泳流速度。这与源自通道壁上的固定电荷和膜附近的感应空间电荷的相反方向的电渗流相结合,为负示踪生物分子的局部预浓缩提供了基础。