Vallejo Derek, Lee Shih-Hui, Lee Abraham
Department of Biomedical Engineering, University of California, Irvine, Irvine, CA, 92697 2715, USA.
Methods Mol Biol. 2017;1572:489-510. doi: 10.1007/978-1-4939-6911-1_31.
In recent years, lipid vesicles have become popular vehicles for the creation of biosensors. Vesicles can hold reaction components within a selective permeable membrane that provides an ideal environment for membrane protein biosensing elements. The lipid bilayer allows a protein to retain its native structure and function, and the membrane fluidity can allow for conformational changes and physiological interactions with target analytes. Here, we present two methods for the production of giant unilamellar vesicles (GUVs) within a microfluidic device that can be used as the basis for a biosensor. The vesicles are produced from water-in-oil-in-water (W/O/W) double emulsion templates using a nonvolatile oil phase. To create the GUVs, the oil can be removed via extraction with ethanol, or by altering the interfacial tension between the oil and carrier solution causing the oil to retract into a cap on one side of the structure, leaving behind an exposed lipid bilayer. Methods to integrate sensing elements and membrane protein pores onto the vesicles are also introduced in this work.
近年来,脂质囊泡已成为构建生物传感器的常用载体。囊泡能够将反应成分容纳在选择性渗透膜内,该膜为膜蛋白生物传感元件提供了理想的环境。脂质双分子层能使蛋白质保持其天然结构和功能,且膜的流动性可允许构象变化以及与目标分析物进行生理相互作用。在此,我们展示了两种在微流控装置中制备巨型单层囊泡(GUVs)的方法,这些囊泡可作为生物传感器的基础。囊泡由水包油包水(W/O/W)双乳液模板使用非挥发性油相制备而成。为了制备GUVs,可通过用乙醇萃取来去除油,或者通过改变油与载体溶液之间的界面张力,使油缩回到结构一侧的帽状物中,从而留下暴露的脂质双分子层。本文还介绍了将传感元件和膜蛋白孔整合到囊泡上的方法。