Development of a virus biosensor platform based on multivalent synthetic nanoswitches

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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.