Luk Vivienne N, Mo Gary Ch, Wheeler Aaron R
Department of Chemistry, University of Toronto, 80 St. George Street, Toronto, Ontario M5S 3H6, Canada.
Langmuir. 2008 Jun 17;24(12):6382-9. doi: 10.1021/la7039509. Epub 2008 May 16.
Digital microfluidics (DMF) is a promising technique for carrying out miniaturized, automated biochemical assays in which discrete droplets of reagents are actuated on the surface of an array of electrodes. A limitation for DMF is nonspecific protein adsorption to device surfaces, which interferes with assay fidelity and can cause droplets to become unmovable. Here, we report the results of a quantitative analysis of protein adsorption on DMF devices by means of confocal microscopy and secondary ion mass spectrometry. This study led us to a simple and effective method for limiting the extent of protein adsorption: the use of low concentrations of Pluronic F127 as a solution additive. This strategy has a transformative effect on digital microfluidics, facilitating the actuation of droplets containing greater than 1000-fold higher protein concentrations than is possible without the additive. To illustrate the benefits of this new method, we implemented a DMF-driven protein digest assay using large concentrations (1 mg/mL) of protein-substrate. The use of Pluronic additives solves a sticky problem in DMF, which greatly expands the range of applications that are compatible with this promising technology.
数字微流控(DMF)是一种很有前景的技术,可用于进行小型化、自动化生化分析,其中试剂的离散液滴在电极阵列表面上被驱动。DMF的一个局限性是蛋白质会非特异性吸附到设备表面,这会干扰分析的保真度,并可能导致液滴无法移动。在这里,我们报告了通过共聚焦显微镜和二次离子质谱对DMF设备上蛋白质吸附进行定量分析的结果。这项研究使我们找到了一种简单有效的方法来限制蛋白质吸附的程度:使用低浓度的普朗尼克F127作为溶液添加剂。这种策略对数字微流控有变革性影响,有助于驱动含有比不添加添加剂时蛋白质浓度高1000倍以上的液滴。为了说明这种新方法的好处,我们实施了一种使用高浓度(1 mg/mL)蛋白质底物的DMF驱动的蛋白质消化分析。普朗尼克添加剂的使用解决了DMF中的一个棘手问题,这极大地扩展了与这种有前景的技术兼容的应用范围。