An Online, Real-Time Microfluidic Biosensor for PFOA and PFOS

适用于 PFOA 和 PFOS 的在线实时微流体生物传感器

• 批准号: 10383822
• 负责人: Nicholas Csicsery
• 金额: $ 25.66万
• 依托单位: QUANTITATIVE BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-01 至 2022-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10383822
• 关键词: Address; Agglutination; Agriculture; Binding; Biological Assay; Biosensing Techniques; Biosensor; Calibration; Detection; Engineering; Ensure; Environment; Escherichia coli; Fluorescence; Generations; Health; Heavy Metals; Home; Human; Image; Individual; Kinetics; Laboratories; Liver diseases; Malignant Neoplasms; Measures; Microfluidic Microchips; Microfluidics; Monitor; Nutrient; Poly-fluoroalkyl substances; Privatization; Process; Quality of life; Risk; Running; Safety; Signal Transduction; Small Business Innovation Research Grant; Specificity; Speed; Surface; Thyroid Diseases; Time; United States; Water; Water Supply; Work; base; commune; drinking; drinking water; epidemiology study; ground water; imaging platform; innovation; microbial; microfluidic technology; nanobodies; novel; novel strategies; operation; prototype; response; sensor; sensor technology; synthetic biology; vector; water testing

Access to clean, reliable water supplies is critical to our quality of life and our economy, and ensuring this access for generations to come will involve developing novel approaches to determining the safety and composition of potable water that are practical and affordable. Per- and polyfluoroalkyl substances (PFASs) are among the most ubiquitous and persistent contaminants plaguing groundwater in the United States, and human epidemiological studies have found associations between PFASs in drinking water and a number of adverse health conditions, from liver and thyroid disorders to various forms of cancer. The main objective of this SBIR proposal is to develop a customizable biosensor platform that uses engineered microbial sensor strains paired with microfluidic technology to continuously monitor water for PFASs. In order to demon- strate technical feasibility, QBI will perform these Specific Aims: Specific Aim 1: To identify and characterize the binding kinetics of nanobodies for PFOA and PFOS. For an engineered bacterial strain to be maximally effective as a sensor, it must be able to specifically and strongly bind its target in the environment. QBI will identify nanobodies that bind PFAS molecules and characterize their capture potential when surface displayed in an adsorbing E. coli strain. QBI will work with a local nanobody company, Abcore, to isolate a set of nanobodies that are enriched for specificity to the two targets and will then bring these nanobodies to their facility, clone them into their E. coli nanobody display vector, and screen and characterize them within their multiplexed microfluidic platform. Specific Aim 2: To develop a prototype for continuous and batch PFAS sensing. In order to use the newly developed sensor strains in a continuous monitoring platform, QBI will need to develop a novel as- say for measuring agglutination on a microfluidic-scale from many individual strain banks. QBI will begin by developing and optimizing an agglutination assay using the reduced set of surface-displayed nanobody strains, and then they will build upon previous results to transduce the agglutination signal to a fluores- cence response. Finally, QBI will optimize a microfluidic device to facilitate this assay and maximize the cellular fluorescence signal so that they can quantify the amount of contaminant present in the water. Successful completion of these Aims will serve to validate the use of nanobody-based sensing strains to achieve sensitive, selective, and continuous contaminant detection, making it of great utility to monitoring efforts aimed at tracking and assessing potential hazardous exposures. Beyond the ability to detect many different targets with a single on-line sensor, which is highly unique, the customizability and expandability of the platform using synthetic biology to engineer strains is transformative. This will enable QBI to continually expand their customer base as they continue to add sensing capabilities tailored to meet end-users' needs. 1

获得清洁、可靠的供水对于我们的生活质量和经济至关重要，并确保 子孙后代的这种获取将涉及开发新方法来确定安全性 实用且经济实惠的饮用水成分。 (PFAS) 是困扰美国地下水的最普遍、最持久的污染物之一 各州和人类流行病学研究发现饮用水中的 PFAS 与 许多不利的健康状况，从肝脏和甲状腺疾病到各种癌症。 该 SBIR 提案的主要目标是开发一个可定制的生物传感器平台，该平台使用工程微生物 传感器菌株与微流体技术相结合，持续监测水中的 PFAS。 为了确定技术可行性，QBI 将实现以下具体目标： 具体目标 1：识别和表征纳米抗体与 PFOA 和 PFOS 的结合动力学。 为了使工程菌株作为传感器发挥最大作用，它必须能够特异性地、 QBI 将识别结合 PFAS 分子的纳米抗体， 表征当表面展示在吸附性大肠杆菌菌株中时它们的捕获潜力。 与当地的纳米抗体公司 Abcore 合作，分离出一组经过富集特异性的纳米抗体 两个目标，然后将这些纳米抗体带到他们的设施，将它们克隆到大肠杆菌纳米抗体中 显示矢量，并在其多路微流控平台中对其进行筛选和表征。 具体目标 2：开发连续和批量 PFAS 传感原型。 在连续监测平台中新开发的传感器菌株，QBI 将需要开发一种新颖的as- 比如说，从许多单独的菌株库中测量微流体规模的凝集反应将从 QBI 开始。 使用减少的表面展示纳米抗体组开发和优化凝集测定 菌株，然后他们将根据先前的结果将凝集信号转导为荧光素 最后，QBI 将优化微流体装置以促进该检测并最大化 细胞荧光信号，以便他们可以量化水中存在的污染物的量。 这些目标的成功完成将有助于验证基于纳米抗体的传感菌株的使用，以实现 灵敏、选择性和连续的污染物检测，使其对于旨在监测目标的工作非常有用 跟踪和评估潜在的危险暴露超出了检测许多不同目标的能力。 单一在线传感器，具有高度独特性，平台的可定制性和可扩展性 使用合成生物学来设计菌株具有变革性，这将使 QBI 能够不断扩展。 他们的客户群不断增加定制的传感功能以满足最终用户的需求。 1