A Paper-Based Synthetic Biology Platform for the On-Demand Testing of Water Quality

项目总结/摘要 安全饮用水对公众健康至关重要，但日益受到人类活动的威胁， 老化的基础设施，重金属和其他毒素污染它。在美国最值得注意的是 饮用水被铅污染，铅是一种普遍存在于美国水基础设施中的重金属 和室内管道长期接触铅会导致许多不利的健康和社会后果， 是一种扩大不平等的环境不公正。一旦发现，铅污染可以通过以下方式减轻： 改变饮用水源，冲洗和现有的过滤技术。然而，由于最近发生的事件， 正如密歇根州弗林特或新泽西州纽瓦克的危机所证明的那样，管理不当可能导致重大公共卫生威胁 通过使整个社区暴露在危险的高水平铅中。饮用水铅污染 由于基础设施老化和家庭管道严重依赖 线索了经常监测饮用水中铅污染的存在是解决方案的一部分。它可以 确定问题所在，及时向消费者发出警报，并告知风险缓解和补救措施 战略布局然而，可靠的测试仍然局限于分析化学技术，这些技术成本高、时间长、 消耗，并需要大量的实验室基础设施和技术专长。这使大规模的- 解决饮用水中的铅危机所需的测试规模，是对水进行常规测试的障碍 消费者的供应。在此，我们建议通过下一阶段的发展来解决这些问题 我们的技术平台，将允许可靠的，廉价的，现场和按需监测铅 在饮用水中。我们的技术是建立在合成生物学的最新创新，使我们能够重新利用 生物传感器蛋白，检测特定的有毒配体，如重金属，进入“无细胞”反应， 当铅存在时产生可检测的信号。这些生化反应是 安全且可嵌入 用于长期存储和分发。向这些纸基设备中加水会激活 生物化学反应，并在有毒化合物的存在下产生视觉信号。这项II期 提案详细说明了一系列补充目标，以提高该电极导线的特异性和灵敏度 测试设备，使用以人为本的设计过程来改善测试操作和结果解释，以及 调整我们的制造和质量保证/质量控制流程，以扩大制造规模 验证研究的能力。一个成功的结果，这一建议将导致一个快速的铅测试与灵敏度 与实验室测试相当，一种可访问且易于使用的设备格式，带有配套的移动的应用程序， 和中试规模生产，以验证我们领先的传感技术在实验室和现场。这项工作 将能够制造和商业销售高度灵敏和快速的铅生物传感器， 帮助解决美国及其他地区日益严重的水质危机。

PROJECT SUMMARY/ABSTRACT Safe drinking water is essential for public health yet is increasingly threatened by anthropogenic activities and aging infrastructure that contaminate it with heavy metals and other toxins. Most notable in the United States is the contamination of drinking water with lead, a heavy metal that is pervasive in America’s water infrastructure and indoor plumbing. Chronic exposure to lead results in numerous adverse health and societal outcomes and is an environmental injustice that widens inequalities. Upon discovery, lead contamination can be mitigated by changing drinking water source, flushing, and existing filtration technologies. However, as recent events such as the crises in Flint, MI or Newark, NJ exemplify, improper management can lead to major public health threats by creating community-wide exposure to dangerously high levels of lead. Lead contamination of drinking water is widespread across the United States due to aged infrastructure and domestic plumbing that heavily relied on lead. Frequent monitoring for the presence of lead contamination in drinking water is part of the solution. It can identify where problems exist, alert consumers without delay, and inform risk mitigation and remediation strategies. However, reliable testing remains limited to analytical chemistry techniques that are costly, time consuming, and require substantial laboratory infrastructure and technical expertise. This complicates the large- scale of testing needed to address the lead in drinking water crisis and is a barrier to the routine testing of water supplies by consumers. Here, we propose to address these issues by pursuing the next stage of development of our technology platform that will allow for the reliable, inexpensive, on-site, and on-demand monitoring of lead in drinking water. Our technology is built from recent innovations in synthetic biology that allow us to repurpose biological sensor proteins that detect specific toxic ligands, such as heavy metals, into ‘cell-free’ reactions that produce detectable signals when lead is present. These biochemical reactions are safe and can be embedded on paper devices for long term storage and distribution. Adding water to these paper-based devices activates the biochemical reaction and produces a visual signal in the presence of a toxic compound. This Phase II proposal details a series of complementary aims for achieving improved specificity and sensitivity of this lead testing device, using a human-centered design process to improve test operation and results interpretation, and adapting our manufacturing and quality assurance/quality control processes for a scale-up of manufacturing capability for validation studies. A successful outcome of this proposal will lead to a rapid lead test with sensitivity comparable to laboratory testing, an accessible and easy-to-use device format with a companion mobile app, and pilot-scale production to validate our lead sensing technology in the laboratory and in the field. This work will enable manufacturing and commercial distribution of a highly sensitive and rapid lead biosensor, which will help address the growing water quality crisis in the United States and beyond.