Deciphering Biomacromolecular Heterogeneity by Single Molecule Mass Spectrometry

Ziel

Virus-like particles, nucleic acids, and other biomacromolecular assemblies have shown promise as vaccine platforms, gene therapy vectors, and more. The molecular composition of these contemporary medicines can be incredibly complicated, with a combinatorial explosion of possible variants (e.g. covalent modifications, isoform substitutions, cargo loads, etc.), that together produce an immense degree of chemical heterogeneity. This is further exacerbated by downstream interactions (e.g. receptors, ligands) which must also engage in a multivalent manner.

Such intrinsic heterogeneity hinders both the rational design of macromolecular assemblies and their detailed molecular characterization. There is a critical unmet need for methodologies capable of accurately characterizing the composition of these pharmaceutically important systems. Mass spectrometry (MS) is a promising technique to meet this need but suffers from poor performance under these high-heterogeneity conditions.

DeMIST will develop new high-resolution single molecule MS methods that will exactly address this limitation and revolutionize the compositional characterization of heterogeneous macromolecules and their interactions. DeMIST will combine instrument and method advancements to enable precise trapping and manipulation of individual molecules, that I envision will overcome existing weaknesses and lead to unprecedented increases in the achievable resolution, sensitivity, and information content of these single ion mass measurements.

DeMIST will yield a unique platform for understanding the composition, assembly, structure, and functional elements of complex (bio)macromolecular assemblies, and an avenue to rationally design and potentially improve this important class of therapies. If successful, DeMIST will allow the scientific community to exploit the full benefits of single molecule mass spectrometry to address previously infeasible biomolecular systems.