Development of an Electroactive Fiber-Optic Chip for Optical/Electrochemical Characterization of Planar-Supported Biological Materials

该合同支持一种新型光纤仪器的开发，该仪器将允许对底物支持的生物分子膜进行宽带、高灵敏度的光谱/电化学表征，包括膜结合蛋白及其电活性成分。提出的仪器是基于一种新的电活性光纤芯片（EA-FOC），它可以提供：1)对表面限制和膜结合的生物相关分子进行超灵敏的吸光度和荧光表征，2)在微升细胞体积下使用10-100飞摩尔的表面限制氧化还原试剂对蛋白质进行光谱/电化学（伏安、安培或电位）表征；3)荧光检测离子通过膜组件运输过程中信号分子环境的变化。该仪器将首次使集成光波导和电活性集成光波导（EA-IOW）技术的灵敏度增强与光纤分光光度计的易用性相结合。计划中的努力扩展了先前的pi工作，以允许检测膜结合生物分子的结构和活性变化，并增加应用范围。该仪器将能够研究结构复杂性不断增加的生物分子膜中表面受限的光学/电化学事件，最终研究重构成平面支撑脂质双层的膜蛋白。EA-FOC的最终设计预计将提供一个可以与广泛的商用仪器集成的平台，并将为开发多功能、敏感和选择性的生物和化学传感器平台提供机会。直接参与该项目的学生将接受新仪器的设计、制造、表征和测试方面的培训，并将获得光谱/电化学技术、光纤技术和光谱学等方面的深入经验，以及将这些工具应用于生物分子膜研究的经验。

This award supports development of a new, fiber-optic-based instrument which will permit broadband, highly sensitive spectro/electrochemical characterization of substrate-supported biomolecular films, including membrane-bound proteins and their electroactive constituents. The proposed instrumentation is based on a new electroactive fiber-optic chip (EA-FOC) that can provide: 1) ultra-sensitive absorbance and fluorescence characterization of surface-confined and membrane-bound biologically relevant molecules, 2) spectro/electrochemical (voltammetric, amperometric or potentiometric) characterization of proteins at microliter cell volumes and using 10-100 femtomoles of surface-confined redox reagent; 3) fluorescence detection of changes in the environment of signaling molecules that occur during ion transport through membrane assemblies. The instrument will make accessible, for the first time, the sensitivity enhancements of integrated optic waveguide and electroactive integrated optic waveguide (EA-IOW) technologies combined with the ease-of-use of fiber-optic based spectrophotometers. The planned effort extends previous work of the PIs to permit detection of changes in the structure and activity of membrane-bound biomolecules and increase the range of applications. The instrument will be able to study surface-confined optical/electrochemical events in biomolecular films of increasing architectural complexity, culminating with studies of membrane proteins reconstituted into planar supported lipid bilayers. The final design of the EA-FOC is expected to provide a platform that can be integrated with a wide range of commercially available instrumentation, and will provide opportunities for the development of versatile, sensitive, and selective bio- and chemical sensor platform. Students directly involved in the project will be trained in the design, fabrication, characterization, and testing of new instrumentation, and will acquire an in-depth experience in several aspects of spectro/electrochemistry techniques, optical fiber technologies, and optical spectroscopies, and in the application of those tools into studies of biomolecular films.