RAPID: Molecular Imprinting of Coronavirus Attachment Factors to Enhance Disinfection by a Selective Photocatalytic “Trap-and-Zap” Approach

在通风管道中发现了导致COVID-19的病毒（即SARS-CoV-2），这表明它可以通过空调系统在建筑物周围传播。尽管患者的检测结果呈阴性，但SARS-CoV-2也会通过粪便排出，因此它可能会到达污水处理厂，在那里它可以存活数天，并被雾化或随废水排放。事实上，有报道称SARS-CoV-2可能通过浴室管道传播。尽管一些传统的水处理工艺可以灭活冠状病毒，但迫切需要更精确、在现实情况下快速、高效和可靠的病毒消毒方法。本项目旨在研究SARS-CoV-2和其他致病性冠状病毒的选择性吸附和光催化消毒（即诱捕和杀灭）新方法。这将导致一种无化学物质的技术（从而避免有害的消毒副产物）以前所未有的精度和可靠的效率灭活冠状病毒。驱动假说是，氮化石墨碳与常见冠状病毒附着因子的分子印迹将使病毒在活性位点附近选择性吸附，从而实现可靠的高消毒。虽然提高废水消毒和医院空气消毒系统的能力和弹性以保护公众健康免受新发传染病的侵害具有显著的内在优点，但本项目的好处要广泛得多。该项目将加强对各种冠状病毒（例如导致COVID-19、中东呼吸综合征和SARS的冠状病毒）的表面识别，这将有助于集中它们，提高精确分离（例如，通过优质吸附剂）和可用于诊断和监测工作的传感器的检测极限。项目成果将整合到各种课程中，包括莱斯大学的教师纳米环境工程（NEET）课程，该课程招收15名教师，每年为3300多名高中生提供服务。这门课程最近扩展到亚利桑那州立大学，也正在扩展到德克萨斯大学埃尔帕索分校，从而确保这种“陷阱和攻击”方法在STEM教师中得到广泛传播。该项目建立在最近发表的一项纳米技术支持的“捕获和破坏”方法（通过分子印迹增强）的基础上，该方法选择性地吸附抗生素抗性基因并将其集中在光催化位点附近进行有效降解（doi.org/10.1021/acs.est.9b06926）。通过将唾液酸、硫酸肝素蛋白多糖、血管紧张素转换酶相关羧肽酶（ACE2）相关肽等参与病毒附着的分子印迹到石墨碳氮光催化剂上，对该方法进行改进，以靶向SARS-CoV-2和其他冠状病毒。当印迹分子被去除（例如，通过酸洗）时，它会留下一个目标特异性的空腔，可以在最小的背景水成分干扰下进行选择性吸附和光催化失活。低致病性冠状病毒HCoV-NL63（与SARS-CoV 2类似，被S刺突蛋白包裹，并使用与宿主受体相同的细胞表面分子ACE2）将用于评估分子印迹gc3n4在竞争蛋白（如牛血清白蛋白）和噬菌体MS2存在下的吸附动力学和选择性。灭活效率将通过定量剩余活病毒浓度来评估，使用Avicel覆盖的空斑试验。具体任务包括：(1)选择冠状病毒模型（如HCoV-NL63）和附着因子进行分子印迹；(2)制备均匀、稳定的生物分子印迹材料；(3)合成分子印迹催化剂；(4)表征目标冠状病毒颗粒对分子印迹催化剂的吸附动力学和选择性；(5)在现实条件下，分子印迹包覆催化剂对传统消毒方法（氯化、紫外线照射）的病毒灭活效率的基准；项目结果将整合到各种课程中，包括莱斯大学的教师纳米环境工程（NEET）课程，该课程每年招收15名教师，覆盖3300多名高中生。这门课程最近扩展到亚利桑那州立大学，也正在扩展到德克萨斯大学埃尔帕索分校，从而确保这种“陷阱和攻击”方法在STEM教师中得到广泛传播。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The virus that causes COVID-19 (i.e., SARS-CoV-2) has been found in air ducts, suggesting that it could spread around buildings via air conditioning systems. SARS-CoV-2 is also shed in stool despite patients testing negative, and thus it may reach wastewater treatment plants, where it could survive for days and be aerosolized or discharged in the effluent. In fact, there are reports that SARS-CoV-2 may spread through bathroom pipes. Although coronaviruses can be inactivated by some conventional water treatment processes, there is an urgent need for more precise viral disinfection approaches that are fast, efficient and reliable under realistic scenarios. The objective of this project is to develop a novel approach for selective adsorption and photocatalytic disinfection (i.e., trap-and-zap) of SARS-CoV-2 and other pathogenic coronaviruses. This would result in a chemical-free technology (thus avoiding harmful disinfection byproducts) with unprecedented precision and reliable efficiency to inactivate coronavirus. The driving hypothesis is that molecular imprinting of graphitic carbon nitride with common coronavirus attachment factors will enable selective virus adsorption near reactive sites, resulting in reliably high disinfection. Whereas enhancing the capacity and resiliency of wastewater disinfection and hospital air sterilization systems to protect public health against emerging infectious diseases has significant intrinsic merit, the benefits of this project are much broader. This project will enhance surface recognition of various types of coronavirus (e.g., those causing COVID-19, MERS and SARS), which will inform efforts to concentrate them and improve both precision separation (e.g., by superior sorbents) and detection limits of sensors that can be used in diagnostics and surveillance efforts. Project results will be integrated into various courses, including the NanoEnvironmental Engineering for Teachers (NEET) course at Rice, which enrolls 15 teachers that reach over 3,300 high school students annually. This course recently expanded to Arizona State University and is also being expanded to the University of Texas at El Paso, thereby ensuring wide dissemination of this “trap-and-zap” approach to STEM teachers.This project builds on a recently published, nanotechnology-enabled “trap-and-zap” approach (enhanced by molecular imprinting), to selectively adsorb antibiotic resistant genes and concentrate them near photocatalytic sites for efficient degradation (doi.org/10.1021/acs.est.9b06926). This approach will be modified to target SARS-CoV-2 and other coronavirus by imprinting molecules involved in virus attachment such as sialic acids, heparin sulfate proteoglycan, and angiotensin-converting enzyme–related carboxypeptidase (ACE2)-associated peptides onto the graphitic carbon nitride photocatalysts. When the imprinted molecule is removed (e.g., by acid washing), it leaves behind a target-specific cavity that enables selective adsorption and photocatalytic inactivation with minimum interference by background water constituents. Low pathogenic coronavirus HCoV-NL63 (which, similarly to SARS-CoV 2, is enveloped with an S spike protein and uses the same cell surface molecule ACE2 as host receptor) will be used to assess adsorption kinetics and selectivity of molecularly imprinted-gC3N4 in the presence of competing proteins (such as bovine serum albumin) and bacteriophage MS2. Inactivation efficiency will be assessed by quantifying residual viable virus concentrations, using the plaque assay with Avicel overlay. Specific tasks include to (1) select a model coronavirus (e.g., HCoV-NL63) and attachment factors for molecular imprinting; (2) prepare homogenous, stable, biomolecular materials for imprinting; (3) synthesize the molecularly-imprinted catalyst; (4) characterize adsorption kinetics and selectivity of target coronavirus particles to the molecularly-imprinted catalyst; (5) benchmark virus inactivation efficiency of the molecularly imprinted -coated catalyst against traditional disinfection methods (chlorination, ultraviolet irradiation) under realistic conditions; and (6) assess durability and reuse potential of the molecularly imprinted catalyst Project results will be integrated into various courses, including the NanoEnvironmental Engineering for Teachers (NEET) course at Rice, which enrolls 15 teachers that reach over 3,300 high school students annually. This course recently expanded to Arizona State University and is also being expanded to the University of Texas at El Paso, thereby ensuring wide dissemination of this “trap-and-zap” approach to STEM teachers.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.