• DCs collectively advanced our understanding of ion-preparation conditions, instrument-dependent resolution, and interpretation of CCS distributions to the structural behaviour of analytes in the gas phase. This work provides the scientific basis for establishing IM-HRMS as a versatile, scalable platform for structural analysis in both academic and industrial settings.
• A coherent framework combining baseline descriptors, artefact-detection rules, and resolution-dependent interpretation, applicable across all molecular sizes was developed.
• Robust IM-HRMS workflows for biotherapeutics, with dedicated developments for oligonucleotide therapeutics, monoclonal antibodies, and viral vectors were established by DCs.
• DCs were trained in handling and preparing biomolecules from various sources, including cancer cells, plasma, sebum, and established protocols for the extraction and/or analysis of proteins, lipids, nucleosides, and virus particles. The acquired skillsets will allow them to generate high-quality biological datasets (lipidomics, proteomics, nucleosides) with IM-HRMS.
• Advanced IM-HRMS methods for nucleoside and cap analysis were developed. Cycling through two different acquisition settings enables both analyte groups to be accommodated in a single measurement.
• DCs used four complementary IM-HRMS technologies (TIMS, DTIMS, TWIMS and SLIM) to harmonise analytical parameters, as well as implement effective QA/QC procedures and data treatment strategies within a vendor-agnostic workflow.
• Establishing a unified and transferable analytical framework for challenging non-targeted screening workflows, strengthening reproducibility and interoperability of datasets is underway.
• Substantial progress in establishing a harmonised temperature calibration framework supported by quantitative energy scaling and cross-platform validation was made.
• Structurally validated and computationally benchmarked collision cross section (CCS) prediction strategies were developed.
• Standardised bioanalytical workflows and a pathway to develop the first reference materials for IM-HRMS have been established.
• Combined efforts toward making calibrated IM-HRMS measurements predictable and quality-controlled across instrument platforms and laboratories are at an advanced stage.
The MobiliTraIN project website is available under www.mobilitrain.eu
