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Microfluidic devices for investigation of biomimetic membranes for sensor and separation applications

机译:用于传感器和分离应用的仿生膜研究的微流体装置

摘要

The term biomimetic membrane denotes membrane that mimics biological cell membrane. Artificially made membranes are powerful tools for the fundamental biophysical studies of membrane proteins. Moreover, they may be used in biomedicine, serving as biosensors in high-throughput screening of potential drug candidates and in separation technologies, where an exciting example is water purification device based on biomimetic membranes containing aquaporins (highly water selective proteins). However, there are many challenges that must be overcome in order to build biomimetic membrane-based devices for industrial applications. Among them are the inherent fragility of lipid membranes, the challenge of up-scaling the effective membrane area and the quantification of the protein delivery to the lipid membrane which may determined the biomimetic membrane application. This PhD thesis addresses the above mentioned difficulties. First, a device that facilitates atomic force microscopy (AFM) measurements of biomimetic membranes is presented. The microfluidic device was specifically designed and fabricated to accommodate the AFM probes that were used to study micrometer-sized fluid polymeric membranes. Second, membrane arrays stability was increased by two ways; surface modification of support partitions and by involving fully closed and automated microfluidic device. The surface was covalently modified by plasma resulting in a hydrophobic coating and thus greatly improved the average membrane array lifetimes (up to 6 days) with a bilayer membrane area ~50% of the available aperture area. Highly stable membranes (up to 2 days) with a bilayer membrane area ~24% of the available aperture area were created in the developed microfluidic device. Further, reconstitution of α-hemolysin (α-HL) membrane proteins in the biomimetic membranes was performed. Third, an outer membrane porin (OMP) fusion efficacy assay was established and used to quantify protein delivery to an array of planar membranes. Incorporation was established as a process with either first order or exponential kinetics. This may be of interest to microfluidic designs involving protein delivery to biomimetic membranes developed for sensor and separation applications. Finally, an OMP functionality modulation with β-cyclodextrin (β-CD) was shown and revealed the protein potential application as a sensor. Moreover, the β-CD blocker may be used to prevent human dental plaque formation and the development of periodontitis.
机译:术语仿生膜表示模仿生物细胞膜的膜。人工制作的膜是用于膜蛋白基础生物学研究的强大工具。此外,它们可用于生物医学中,用作潜在药物候选物的高通量筛选和分离技术中的生物传感器,其中一个令人振奋的例子是基于包含水通道蛋白(高度水选择性蛋白)的仿生膜的净水设备。但是,要构建用于工业应用的仿生膜基设备,必须克服许多挑战。其中包括脂质膜的固有易碎性,扩大有效膜面积的挑战以及定量蛋白质传递至脂质膜的挑战,这可能决定了仿生膜的应用。本博士论文解决了上述困难。首先,提出一种促进仿生膜原子力显微镜(AFM)测量的装置。微流控设备经过专门设计和制造,可容纳用于研究微米级流体聚合物膜的AFM探针。第二,膜阵列的稳定性通过两种方式提高:支撑隔板的表面改性,以及涉及全封闭和自动化的微流控设备。通过等离子体对表面进行共价修饰,形成疏水涂层,从而极大地提高了平均膜阵列寿命(长达6天),双层膜面积约为可用孔径的50%。在开发的微流控设备中创建了双层膜面积约为可用孔径面积约24%的高度稳定的膜(长达2天)。此外,在仿生膜中进行了α-溶血素(α-HL)膜蛋白的重构。第三,建立了外膜孔蛋白(OMP)融合功效测定法,并用于量化蛋白质向平面膜阵列的递送。建立结合是具有一级动力学或指数动力学的过程。这对于涉及将蛋白质递送至为传感器和分离应用开发的仿生膜的微流体设计可能是感兴趣的。最后,显示了用β-环糊精(β-CD)进行的OMP功能调节,并揭示了蛋白质作为传感器的潜在应用。此外,β-CD阻滞剂可用于预防人牙菌斑的形成和牙周炎的发展。

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