SBIR Phase I: Optical Enzymatic Biosensor for Real-Time, Continuous, In-Situ Measurements of Trichloroethene

这个小企业创新研究（SBIR）第一阶段项目提议开发一种无细胞光学酶生物传感器，用于监测水中三氯乙烯（TCE）的浓度。生物传感概念将以光纤形式实现，其中光学生物传感器尖端与一个小型光电单元相连。化学检测发生在涂有酶和荧光化学物质的生物传感器尖端上。在初步实验中，使用含有所需酶的整个细胞的传感器被证明可以测量TCE浓度。然而，这些全细胞传感器在检测限和可制造性方面存在局限性。因此，该项目的研究目标是通过提取必要的多组分酶并将酶结合到生物传感器尖端来开发这种生物传感器的无细胞版本。这些目标的实现将推动TCE传感技术向强大和易于制造的系统发展，用于TCE监测，具有更高的检测限，更高的准确性和可靠性。这个项目如果成功，其更广泛的影响/商业潜力将是提高查明、绘制地图和修复受三氯乙烯污染的水源的效率。用水量的增长速度是人口增长速度的两倍多。在许多国家，水量和水质都成为主要问题。由于水相TCE在水源中的广泛存在、毒性和高稳定性，监测其浓度的必要性将持续许多代。对于能够快速、准确和廉价地测量水中三氯甲烷和其他氯化有机化学品浓度的设备，世界范围内存在着并将继续增加的巨大需求。在环境传感领域持续的现场数据生产将大大提高需要测量TCE浓度的所有过程的效率。这种传感器将允许在污染事件的情况下对可能的污染源进行连续监测。此外，它将提供含水层中TCE羽流的深度和空间剖面。该技术将取代目前基于色谱的测量方案，这些方案不容易适应原位、在线或现场测量，并且不能连续产生数据。

This Small Business Innovation Research (SBIR) Phase I project proposes to develop a cell-free optical enzymatic biosensor for monitoring trichloroethene (TCE) concentrations in water. The biosensing concept will be implemented in an optical fiber format, in which optical biosensor tips are linked to a small electro-optical unit. Chemical detection takes place on biosensor tips that are coated with enzymes and fluorescent chemicals. In preliminary experiments, sensors using whole cells containing the desired enzymes were shown to measure TCE concentrations. However, those whole-cell sensors had limitations with respect to their detection limits and manufacturability. Thus, the research objectives of this project are to develop a cell-free version of this biosensor by extracting the necessary multicomponent enzyme and incorporating the enzyme onto the biosensor tips. Attainment of these objectives will advance the TCE-sensing technology toward a robust and readily manufactured system for TCE monitoring with improved detection limits, increased accuracy, and reliability. The broader impact/commercial potential of this project, if successful, will be to increase the efficiency of identifying, mapping, and remediating water sources contaminated with TCE. Water use has been growing at more than twice the population rate. Both water quantity and water quality are becoming dominant issues in many countries. The necessity of monitoring aqueous-phase TCE concentrations will continue for many generations due to its widespread occurrence in water sources, its toxicity, and its high stability. A significant need exists and will continue to rise worldwide for devices that can measure the concentration of TCE as well as other chlorinated organic chemicals in water, and that can do so rapidly, accurately, and inexpensively. Continuous in-situ data production in the field of environmental sensing will greatly increase efficiency in all processes that require a measurement of TCE concentration. This sensor will allow for continuous monitoring of possible contamination sources in the case of a contamination event. Also, it will afford facile depth and spatial profiling of TCE plumes in aquifers. This technology will replace the current chromatography-based measurement protocols, which are not readily adapted for in-situ, on-line, or in-the-field measurements, and cannot produce data continuously.