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Rapid and in-depth proteomic profiling of small extracellular vesicles for ultralow samples

ABSTRACT: The form and function of extracellular vesicles (EV) is defined by their proteome. This knowledge is essential to describe and understand EVs, encompassing their marker proteins, capacity as a signalling platform, and utility as diagnostic tools and therapeutic targets. However, EV are low-abundant entities in the secreteome and biological samples, with disease-specific EV proteins often present in low abundance challenging their detection and quantification due to the inherent challenges in dynamic range using mass spectrometry-based strategies. Combined with lack of protein amplification mechanisms, their proteomic studies require upscaling cell cultures or larger volumes of biofluids. Here, we outline high-sensitivity sample preparation and finely tuned LC gradients for DIA to obtain precise and comprehensive proteome of EVs from ultra-low sample amounts (sub-nanogram).

INSTRUMENT(S): Q Exactive HF-X

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: David Greening

PROVIDER: MSV000092668 | MassIVE | Wed Aug 16 06:57:00 BST 2023

REPOSITORIES: MassIVE

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