Development of a virus biosensor platform based on multivalent synthetic nanoswitches

基于多价合成纳米开关的病毒生物传感器平台的开发

• 批准号: 464980137
• 负责人: Dr. Alexander Gräwe
• 金额: --
• 依托单位: Department of Biomedical Engineering
• 依托单位国家: 德国
• 项目类别: WBP Fellowship
• 财政年份: 2021
• 资助国家: 德国
• 起止时间: 2020-12-31 至 2021-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/464980137?language=en
• 关键词: Development; virus; biosensor; platform; based

The SARS-CoV-2 pandemic has demonstrated the importance of reliable test pipelines for virus infections. Fast identification of individuals who carry infectious viruses would help to counteract uncontrolled spreading. Although existing biosensing technologies are currently on the rise, these are still limited in terms of speed, read-out capacities and re-usability. Therefore, there is pressing need for fast, user-friendly molecular devices for point-of-care diagnostics. Such reliable and easy-to-use devices will both prove useful as a countermeasure for future epidemics but will also help society counter other harmful diseases. It is highly desirable to achieve direct target detection in solution, as this greatly reduces sample preparation and detection time. Such homogenous assays require conceptual designs that transform a molecular interaction or binding event into a detectable output. This can only be achieved if we learn to understand, predict, and control the behaviour of the respective molecular design. In this regard, recent progress in the field of synthetic biology has yielded advanced biosensors based on modular synthetic protein switches. Yet, the laborious optimization of these switches – and in particular, their internal linkers – has limited their potential for point-of-care applications. The herein proposed project addresses this issue by developing novel auto-inhibited nanoswitches based on multivalent interactions with high sensitivity for the detection of viruses and virus-like targets. Sophisticated linker screening technologies will be employed to arrive at optimal switch architectures. Two complementary types of nanoswitch scaffolds – based on DNA and protein linkers – will be pursued, and both will be evaluated using the trimeric S Glycoprotein from SARS-CoV-2 and Influenza A virus-like particles as analytes. Thus, the main objectives of the project are 1) engineering and construction of these switches and 2) evaluation of the underlying design principles. In addition to yielding a new class of multivalent nanoswitches that allow highly sensitive, 1-step viral detection directly in solution, the fundamental insights gained in multivalent molecular interactions with viral surfaces may also help in the development of novel vaccines and virus neutralisation efforts.

SARS-CoV-2大流行证明了可靠的病毒感染检测管道的重要性。快速识别携带传染性病毒的个人将有助于遏制不受控制的传播。虽然现有的生物传感技术目前正在兴起，但在速度、读出能力和可重复使用性方面仍然有限。因此，迫切需要快速、用户友好的分子装置用于即时诊断。这种可靠和易于使用的设备将被证明是未来流行病的有效对策，也将有助于社会对抗其他有害疾病。非常希望在溶液中实现直接目标检测，因为这大大减少了样品制备和检测时间。这种均相测定需要将分子相互作用或结合事件转化为可检测输出的概念设计。只有当我们学会理解、预测和控制各自分子设计的行为时，才能实现这一点。在这方面，合成生物学领域的最新进展已经产生了基于模块化合成蛋白质开关的先进生物传感器。然而，这些开关的费力优化-特别是它们的内部连接器-限制了它们在护理点应用中的潜力。 本文提出的项目通过开发基于多价相互作用的新型自抑制纳米开关来解决这个问题，该纳米开关具有高灵敏度，用于检测病毒和病毒样靶标。将采用先进的接头筛选技术来达到最佳的开关架构。两种互补类型的纳米开关支架-基于DNA和蛋白质接头-将被追求，并且两者都将使用来自SARS-CoV-2的三聚体S糖蛋白和甲型流感病毒样颗粒作为分析物进行评估。因此，该项目的主要目标是：1）这些开关的工程和施工; 2）基本设计原则的评估。除了产生一类新的多价纳米开关，允许在溶液中直接进行高灵敏度的一步病毒检测外，在多价分子与病毒表面相互作用中获得的基本见解也可能有助于开发新型疫苗和病毒中和工作。