Advances in biotechnology, pharmaceutical and life sciences require improved performance of even the most powerful analytical techniques to target the extreme complexity of modern biological samples. Due to its high performance, Fourier transform mass spectrometry (FTMS) is the central analytical technique in biomolecular analysis. Fourier transform (FT) drives FTMS by converting the time-domain (transient) data to mass spectra which, in turn, provide the biological information. Although FT is robust, it is inherently slow due to its strict uncertainty principle. Thus, many life sciences applications of FTMS are suffering from a limited throughput – data acquisition in FTMS should be done faster! Recent innovation results of our ERC Starting Grant “Super-resolution mass spectrometry for health and sustainability” have revealed the incredible power that methods of advanced signal processing, whose uncertainty principles are less strict than the FT one, have to offer to the everyday routine high-performance FTMS. Thus, we have rationally implemented existing and developed novel super-resolution and advanced signal processing methods to substantially speed up FTMS. While fundamental and technical feasibilities of our approach have been evaluated favorably at the lab level, turning these research outputs into a commercial proposition is yet to be demonstrated. Therefore, the aim of Time2Life is to translate our technology validated in lab into a robust industry-grade technology that accelerates high-performance biological mass spectrometry via the advanced signal processing of time-domain (transient) data and thus leverages life sciences applications. The industrial and academic end-users would be able to identify and quantify more analytes (e.g., peptides, proteins and metabolites) and thus enhance biological significance and accuracy of their research and clinical work. Notably, we target the unrepresented by SME area of time-domain data analysis in mass spectrometry.

Biotherapeutics, such as monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs), constitute an increasingly important class of molecular drugs. Accelerating our society’s access to novel biotherapeutics-based therapies and diagnostic tests for existing and new clinical indications, including those caused by COVID-19, is critical. The exceptional specificity and selectivity of mass spectrometry (MS) renders this analytical technique a powerful tool in development of biotherapeutics-based drugs and clinical diagnostics. Nevertheless, even Orbitrap Fourier transform mass spectrometry (FTMS), which dominates the high-performance MS field, may demonstrate limited performance in structural analysis of the very complex mAbs and ADCs. Indeed, while the user-accessible Orbitrap data (processed mass spectra with reduced information) may be of a sufficient quality and information content for small molecule and isolated proteins analysis, it is not for the complex biotherapeutics. We identified the limiting factor that inhibits biotherapeutics structure analysis being the absence of hardware and software tools to acquire (access and provide to the end-users) and analyze the un-reduced data (e.g., time-domain signals or transients). With these tools in hands, the end-users would be able to increase performance and productivity of their biopharma workflows on the Orbitraps (e.g., leading to increased sensitivity and resolution, and thus leading to faster identification of more species and their deeper sequencing), as well as create novel workflows, not possible otherwise. The A2MStools (access & analysis MS tools) aims to address this limitation by: (i) providing access to the currently unavailable un-reduced data (transients) for the Orbitraps FTMS platforms with enabled BioPharma capabilities, and (ii) empowering analytical scientists with the data analysis tools capable of handling the datasets of unprocessed data in the most comprehensive and innovative ways.

The unbeatable performance leaders in molecular analysis of biological and environmental samples are the Fourier transform mass spectrometers (FTMS): Orbitrap FTMS and ion cyclotron resonance (FT-ICR MS). However, even FTMS approaches suffer from performance limitations when facing the molecular complexity of samples in modern applications. The primary reasons for these limitations are the space charge effects – strong Coulombic interactions of many ions oscillating in the trapping devices. Applications suffering from space charge affects include those with (i) significant fluctuation of incoming ion currents; (ii) close spacing of molecular mass/charge ratios to be resolved; and (iii) high spectral dynamic range of ion intensities. To address these performance challenges and as a result of the completed ERC Starting Grant, we propose to increase precision of mass measurements in FT-ICR MS by using the direct cyclotron frequency measurements instead of the space charge-sensitive reduced cyclotron frequency measurements employed currently. We have confirmed the principal feasibility of FT-ICR MS at the cyclotron frequency on both types of commercial FT-ICR MS instruments on the lab level. The aim of our PoC grant application is to realize the benefits of FT-ICR MS at the cyclotron frequency by developing and seamlessly integrating the laboratory-level innovation into commercial instruments at an industry-grade level to leverage life and environmental sciences applications. The action plan includes: (i) enabling integration of prototype ICR mass analyzers into commercial FT-ICR MS instruments via feasibility studies and pilot projects; (ii) analyzing competitive advantages of the project’s outcomes over existing products and workflows, with a focus on frequency (mass) precision increase; (iii) clarifying IPR position, strategy; and (iv) performing market analysis.

The extraordinary ability of the coronavirus to spread in the environment has established the Covid-19 pandemic as the biggest challenge humanity faces in the XXI century. Covid-19 attacks humans regardless of age, claiming the life of over 8,000 people in a single day, with a devastating death toll exceeding 350,000 in just a few months. The virus is likely to survive for the foreseeable future and disperse further, requiring long-term planning and investment in developing means of detection, protection and cure. Protective measures based on monitoring the dispersion of the coronavirus in the environment and fast screening of individuals has become paramount for ensuring safety of our ageing population and restarting/supporting the worldwide economy. The objective of the ARIADNE project is to develop a multiple-stage analytical platform based on multi-dimensional mass spectrometry instrumentation. Performing direct and instant detection of intact virus particles in breath and in water, and going far beyond that task, this unique analytical platform will push the scientific boundaries in all aspects of analytical sciences centered on mass spectrometry, incorporating a series of potentially disruptive technologies integrated into a single system. ARIADNE integrates state-of-the-art technological advancements in breath sampling and post-ionization methods, new analytical tools for characterization of the protein content of viruses by top-down mass spectrometry, non-destructive ultra-high mass analysis of single particles followed by their soft landing and further processing based on advanced single proteomic workflows. A compact and simplified version of this versatile and powerful analytical platform is also envisaged for advancing the field of real-time breath analytics. Applications extending the analytical capabilities of the system to new viruses, intact bacteria as well as whole human cells will also become accessible.