Alpha Prototyping and Deployment of a MIP-Based Microfluidic Fluorometric Bacteria Biosensor

基于 MIP 的微流控荧光细菌生物传感器的 Alpha 原型设计和部署

• 批准号: 580282-2022
• 负责人: Rezai, PouyaP
• 金额: $ 1.82万
• 依托单位: York University
• 依托单位国家: 加拿大
• 项目类别: Alliance Grants
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=760051
• 关键词: Alpha; Prototyping; Deployment; MIP; Based

Under NSERC and MITACS projects since 2020, in partnership with Sixth Wave Inc. (SIXW), we have developed and patented facile approaches to coat bacterium- and bacteriophage-specific Molecularly Imprinted Polymers (MIPs) on florescent magnetic microparticles (MMPs) and microwires (MWs) as biosensing interfaces. MIPs are synthetic polymers that are prepared in the presence of bio-targets such as bacteria. After engraving the bacteria out with an etchant, there will be cavities specific to the bacteria left in the MIP structure for rebinding to the same target. We used microfluidics to capture MIP-coated MMPs in a microchannel using an external magnet, then fluorescent microscopy was used for proof-of-principle E. coli detection. For this, water samples with bacteria were exposed to the captured MIP-coated MMPs in the device, and fluorescent attenuation of the particles due to bacteria shielding was used to achieve a limit of detection (LOD) of 100 CFU/mL.In this application, one postdoc will be trained to develop a portable fluidic pump and a portable fluorescent microscope, then integrate them with the above microfluidic biosensor to develop an alpha prototype of the sensor. We aim to test the prototype with 4 types of bacteria and characterize its performance such as limit of detection and dynamic range. Our international collaborator is Prof. Adela Ben-Yakar from the University of Texas Austin who will bring expertise in developing high-end imaging and microscopy units. She will provide guidance to the postdoc on a biweekly basis and also may receive the postdoc in her lab for 1-2 months to test the prototype in the USA lab and with some water samples there. Outcomes include 1) Development of a portable platform technology for the detection of multiple bacteria; 2) Training of one postdoc in the biosensor sector; 3) Initiating an international collaboration to bring needed expertise to the team, valuable experience for the postdoc, and unique testing opportunities for the platform; 4) Completion of data for a patent that is currently filed provisionally; 5) Publication of one new journal paper and completion of two existing manuscripts.

自2020年起，在NSERC和MITACS项目下，与Sixth Wave Inc.（SIXW），我们已经开发并获得专利的简易方法，以涂层细菌和噬菌体特异性分子印迹聚合物（MIP）上的flocculant磁性微粒（MMP）和微丝（MW）作为生物传感接口。MIP是在生物靶标如细菌存在下制备的合成聚合物。在用蚀刻剂将细菌雕刻出来之后，在MIP结构中将留下细菌特异性的空腔，用于重新结合到相同的靶标。我们使用微流控技术在微通道中使用外部磁体捕获MIP包被的MMPs，然后使用荧光显微镜进行原理验证E。大肠杆菌检测为此，将含有细菌的水样暴露于装置中捕获的MIP涂覆的MMP，并使用由于细菌屏蔽而导致的颗粒的荧光衰减来实现100 CFU/mL的检测限（LOD）。在该应用中，将培训一名博士后以开发便携式流体泵和便携式荧光显微镜，然后将它们与上述微流体生物传感器集成以开发传感器的α原型。我们的目标是测试原型与4种类型的细菌和表征其性能，如检测限和动态范围。我们的国际合作者是来自德克萨斯大学奥斯汀分校的Awesley Ben-Yakar教授，他将带来开发高端成像和显微镜单元的专业知识。她将每两周为博士后提供指导，也可能在她的实验室接受博士后1-2个月的测试原型在美国实验室和一些水样。成果包括：1）开发用于检测多种细菌的便携式平台技术; 2）在生物传感器领域培训一名博士后; 3）启动国际合作，为团队带来所需的专业知识，为博士后提供宝贵的经验，并为平台提供独特的测试机会; 4）完成目前临时申请的专利的数据; 5）发表一篇新的期刊论文，完成两篇现有手稿。