HIGHLY SELECTIVE MACROBIOMOLECULAR ISOLATION, SOFT-LANDING AND CHARACTERIZATION USING STRUCTURES FOR LOSSLESS ION MANIPULATIONS

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PROJECT SUMMARY/ABSTRACT (30 lines) This project aims to address the current gap in technology for rapid and comprehensive (mass, structure) profiling of biomacromolecules. While high resolution ion mobility and mass spectrometry separations techniques have been extensively developed, their application has been largely limited to small molecules (m/z < 2000). The ability to access structure and mass information in a high-throughput fashion for macrobiomolecules is an aspirational goal in mass spectrometry. In pursuance of this goal, we will develop a high-resolution ion mobility chemical analysis device based on Structures for Lossless Ion Manipulations, and in conjunction with mass spectrometry obtain high-throughput high-resolution IMS/MS of biomacromolecules. The study of the significant heterogeneity in large biomolecular systems, i.e., macrobiomolecules, critical to biological research will be enabled by the development of a new high resolution, high throughput analytical system. The SLIM platform, currently enabled for ultra-high resolution ion mobility profiling (up to the point of separating isotopomers and isotopologues) and manipulations of small molecular ions (<3000 Daltons) will be optimized to efficiently confine, transport and separate macrobiomolecules. Initial optimization of the SLIM will be done by coupling with a TOF-MS system. SLIM-TOF will be used to perform high resolution IMS/MS of ensembles of macrobiomolecular species (with an upper mass limit of about 20,000 Da dictated by the TOF). We will use SLIM-based IMS separations and other manipulations to reveal the extensive structural heterogeneity of macrobiomolecules. Further, we will perform size and mass selective soft-landing of macrobiomolecules using the dynamic switching and rerouting (to deposition substrates) of a selective ion mobility distribution in the SLIM. The soft-landed species will be characterized using cryo-EM and complimentary imaging techniques to elucidate the morphologies of macrobiomolecules with selected mobilities or collision-cross-sections. The combination of ion mobility information and imaging will provide a detailed structural characterization of macrobiomolecules.