UNS: Role of Reporter and Membrane Properties for the Sensitivity of Viral Nanoparticle Lateral Flow Assays

UNS：报告基因和膜特性对病毒纳米颗粒侧向层析检测灵敏度的作用

• 批准号: 1511789
• 负责人: Jacinta Conrad
• 金额: $ 32.96万
• 依托单位: University of Houston
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-15 至 2020-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1511789&HistoricalAwards=false
• 关键词: UNS; Role; Reporter; Membrane; Properties

CBET - 1511789 Conrad, Jacinta The investigators have shown that significantly improved sensitivity can be achieved by using engineered bio-active nanoparticles. The applications of the proposed method development, if successful, will simplify pathogen detection in a variety of contexts, including food, water and medical samples.The objective of the proposed research is to identify, understand, and ultimately engineer the properties of phage reporters and membranes that provide sensitive response in lateral flow assays (LFA). The hypothesis is tailoring the properties of phage nanoparticles and LFA membranes will enhance phage transport and binding and as a result reduce LFA limit of detection. The specific aims are to: (1) vary phage morphology to understand phage performance as LFA reporters; (2) identify membrane properties that enhance phage transport and adhesion onto the membrane; and (3) select phage for superior performance using directed evolution. Phage reporters of varied shape and size will be imaged with confocal microscopy as they diffuse through and bind to LFA membranes. Using high-throughput image-processing algorithms, phage orientation and dispersion, binding will be correlated to membrane properties and pore size. Directed evolution will be used to select phage with improved transport and specificity from libraries of billions of candidates. Intellectual Merit: This project integrates the expertise of the PI in imaging and confined transport with that of the co-PI in chromatographic interactions and phage engineering to develop LFAs of increased sensitivity. Phage reporters are potentially transformative as LFA reporters, enabling fast, inexpensive and ultrasensitive detection with extremely low non-specific binding. The proposed research will establish ultrasensitive LFAs as a broadly-applicable platform technology. The transport of nonspherical nanoparticles in porous media is under-investigated, and the proposed work will use phage as a new model system in for studing dispersion in complex and confined media. Finally, this proposal introduces phage evolution with separation as a novel method to engineer bacteriophage for applications in the emerging area of nanobiotechnology. Broader Impacts: Viral nanoparticle LFA technology can be readily integrated with microfluidics and smartphone-based fluorescence imaging to yield an inexpensive yet ultrasensitive point-of-use diagnostic tool. Bioseparations dominate the cost and process complexity of manufacturing of modern biopharmaceuticals; process analytical (PAT) technologies that do not require trained investigators will immediately impact this area. In addition, a field-portable analytical method may prove useful for environmental contamination, food safety monitoring, and medical diagnostics. The University of Houston is the second-most ethnically diverse research university in the US and is designated as a Hispanic-serving Institution, providing a fertile recruiting ground for broadening participation from underrepresented groups. The PIs will continue ongoing outreach efforts aimed at the local scientific community, by organizing the Texas Soft Matter Meeting for academic and industrial scientists, and at society at large, by writing and presenting popular science radio programs for The Academic Minute and Engines of our Ingenuity.

CBET - 1511789 Conrad, Jacinta 研究人员表明，通过使用工程生物活性纳米颗粒可以显着提高灵敏度。如果成功的话，所提出的方法开发的应用将简化各种情况下的病原体检测，包括食品、水和医学样本。拟议研究的目的是识别、理解并最终设计噬菌体报告基因和噬菌体报告基因的特性。在侧流分析 (LFA) 中提供灵敏响应的膜。 该假设是定制噬菌体纳米颗粒和 LFA 膜的特性将增强噬菌体运输和结合，从而降低 LFA 的检测限。具体目标是：（1）改变噬菌体形态以了解噬菌体作为 LFA 报告基因的性能； (2) 确定增强噬菌体转运和粘附到膜上的膜特性； (3) 使用定向进化选择具有优越性能的噬菌体。当不同形状和大小的噬菌体记者扩散穿过 LFA 膜并与其结合时，将通过共聚焦显微镜对其进行成像。使用高通量图像处理算法、噬菌体定向和分散、结合将与膜特性和孔径大小相关。定向进化将用于从数十亿候选库中选择具有改善的运输和特异性的噬菌体。 智力优势：该项目将 PI 在成像和限制运输方面的专业知识与 Co-PI 在色谱相互作用和噬菌体工程方面的专业知识相结合，以开发灵敏度更高的 LFA。噬菌体报告基因与 LFA 报告基因一样具有潜在的变革性，能够以极低的非特异性结合实现快速、廉价和超灵敏的检测。拟议的研究将把超灵敏的 LFA 建立为一种广泛适用的平台技术。非球形纳米粒子在多孔介质中的传输尚未得到充分研究，拟议的工作将使用噬菌体作为新的模型系统来研究复杂和受限介质中的分散。最后，该提案介绍了噬菌体进化与分离作为一种改造噬菌体的新方法，以应用于纳米生物技术的新兴领域。 更广泛的影响：病毒纳米粒子 LFA 技术可以轻松地与微流体和基于智能手机的荧光成像集成，以产生一种廉价但超灵敏的使用点诊断工具。生物分离在现代生物制药制造的成本和工艺复杂性中占主导地位；不需要训练有素的调查人员的过程分析（PAT）技术将立即影响这一领域。此外，现场便携式分析方法可能对环境污染、食品安全监测和医疗诊断有用。休斯顿大学是美国第二大种族多元化的研究型大学，被指定为拉美裔服务机构，为扩大代表性不足群体的参与提供了肥沃的招聘土壤。 PI 将继续针对当地科学界开展外展工作，为学术和工业科学家组织德克萨斯州软物质会议，并为整个社会编写和播放《学术分钟和我们创造力的引擎》的科普广播节目。