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Redefining mass spectrometry – a breakthrough platform for real-time noninvasive breath analysis with single ion detection of intact viruses and bacteria and post-analysis molecular characterization

Periodic Reporting for period 3 - ARIADNE (Redefining mass spectrometry – a breakthrough platform for real-time noninvasive breath analysis with single ion detection of intact viruses and bacteria and post-analysis molecular characterization)

Reporting period: 2023-12-01 to 2025-02-28

The objective of the ARIADNE project is to develop a breakthrough analytical platform based on mass spectrometry (MS) for detecting intact virus particles directly from breath. An entire new range of highly disruptive technologies are currently being realized. These technologies involve the development of novel sampling interfaces and complex aerosol processing methods, new ionization sources for supercharging particles and droplets, gas dynamically optimized transfer optics for very high m/z ions, advanced top-down analytical workflows in the ultra-high mass range and mass analyzer designs with integrated novel detection schemes. During the course of the project, breakthrough results have been produced on single-bacteria proteomics and new modes of operating Charge-Detection Mass Spectrometry. Detection of ions in the ultra high/mass range has also been made possible using the ARIADNE platform, while novel ionization and breath sampling schemes are being tested.
WP1 - The first operational aerosol supercharger, which now integrates an inertial cutoff filter, has been developed (after three iterations), and fully characterized. The majority of the deliverables has been completed, although it was found that the Limit of Detection of the system is still insufficient to detect viral particles in breath in real time, nevertheless, nm-size particles mimicking intact virus particles produced by ESI have been detected using the ARIANE platform. The breakthrough result on super-charging of aerosol particles is yet to be accomplished. A summary of the efforts undertaken until this third Reporting Period is provided in the Technical Report

WP2 The ARIADNE platform is currently producing mass spectra of ions in the higher m/z range using the conventional detectors. The new electronics to drive the eTOF mode have been produced and tested, while experiments are underway. Preliminary results based on the prototype diffusion charger - FAIMS have been produced and proton transfer reactions are currently being evaluated under different experimental conditions

WP3 - A working prototype is currently being operated in pass-through mode successfully, an updated version of the acquisition system & allied data processing tools has also been developed. A High-Performance Data Acquisition System for CDMS and the next-gen firmware and software for CDMS data acquisition, processing, and analysis have been produced and benchmarked successfully using the CDMS platform in Lyon. Firmware developments and software updates resulted in novel workflows enabling viral particles data simulations.

WP4 - The Omnitrap platform is operational on the QE UHMR mass spectrometer, while first data on analyzing virus particles using this system are produced. Now that the Omnitrap is fully functional, partner’s UzL main focus is operating the QE UHMR in the ultra-high mass-range, which is a necessary prerequisite for performing the top-down experiments combining the Omnitrap technology. In addition, partner FAST will concentrate on the top-down experiments. Finally, partner CNRS-Lyon has redesigned the FT-CDMS to enhance trapping efficiency and execute top-down experiments with the CO2 laser.

WP5 - Partner KI has completed successfully the work package concerning the advancement of single bacterial cell proteomics, a novel proteomics workflow that resulted in a peer review publication. Furthermore, UzL has successfully completed the development of proteomics workflows for viruses. This know-how will now be extended to advance proteomics workflows for analyzing viruses, a very challenging task due to the extremely low number of proteins present in the sample.

WP6 - The Project Management tasks are being implemented successfully, without deviation

WP7 - The Innovation Radar has been updated, steps are being made for the exploitation plan regarding the new technologies developed during the project.
An outstanding list of new technologies is currently being developed within the framework of the project, and successful implementation of only a part of those ideas currently brought forward will be a great success. In this first stage of the project, different subsystems of the instrument are being developed independently by the partners.

ARIADNE is a new radical approach to monitor viral particles exhaled directly from breath in real-time. Successful implementation and execution of the project will lay the foundations for a new direction in the development of high-end analytical instrumentation deployed to fight and contain epidemics and pandemics. Any positive result in the direction of providing fast, sensitive, real-time information on the health state of an individual by tracing the viral load directly from breath will be communicated and set the scene for future research in this direction. The resulting technology will be key to rapidly screen and diagnose large populations. In that regard, the scope of the ARIADNE project remains extremely relevant and the expected impact quite substantial.
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