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

自2020年以来，在NSERC和MITACS项目下，我们与Sixth Wave Inc.(SIXW)合作，开发了一种简便的方法，将细菌和噬菌体特异的分子印迹聚合物(MIP)覆盖在荧光磁性微粒(MMP)和微丝(MWs)上作为生物传感界面，并获得了专利。MIPS是在细菌等生物靶标存在的情况下制备的合成聚合物。用蚀刻剂将细菌雕刻出来后，MIP结构中将留下细菌特有的空洞，用于重新结合到相同的靶标。我们使用微流控技术利用外部磁铁在微通道中捕获MIP包裹的MMPs，然后使用荧光显微镜进行原理验证的大肠杆菌检测。为此，将含有细菌的水样暴露在设备中捕获的MIP涂层MMPs中，并利用细菌屏蔽对颗粒的荧光衰减来实现100cfu/ml的检测极限(LOD)。在这项应用中，一名博士后将接受培训，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控生物传感器集成，开发出传感器的阿尔法原型。我们的目标是用4种细菌对样机进行测试，并对其检测极限和动态范围等性能进行表征。我们的国际合作者是德克萨斯大学奥斯汀分校的Adela 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。她将每两周向博士后提供指导，还可能在她的实验室接受1-2个月的博士后指导，以便在美国实验室测试原型，并在那里采集一些水样。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1名博士后；3)启动国际合作，为团队带来所需的专业知识、博士后宝贵经验和平台独特的测试机会；4)完成目前临时提交的一项专利的数据；5)发表一篇新的期刊论文，并完成两篇现有的手稿。

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.