Highly efficient TiO2-based one-step strategy for micro volume plasma-derived extracellular vesicles isolation and multiomics sample preparation

Extracellular vesicles (EVs) released from tumor cells and tissues mediate intercellular communication and regulate various important biological processes. The characterization of EVs can provide potential diagnostic biomarkers and therapeutic targets. However, current EV enrichment and sample preparation methods for omics analysis are time-consuming and costly, and a relatively large amount of samples is often required. Here, we report a robust TiO2-mediated multiomics EV preparation method (TiMMEP) for isolating EVs from a small volume of plasma and profiling proteins and metabolites simultaneously. Plasma EVs were initially captured by TiO2, and EVs proteins and metabolites were released by adding 70% acetonitrile. Subsequently, the proteins were precipitated and adsorbed on the surface of TiO2 and the metabolites were collected in the supernatant. More than 700 protein groups and 491 tentatively annotated metabolites were identified in the EVs isolated from 1 mL of plasma. Furthermore, the TiMMEP method was applied for multi-omics EV analysis of ten healthy human plasma samples to demonstrate its feasibility and stability. Thus, the TiMMEP strategy enables a highly sensitive, operationally simple, and high-throughput multiomics analysis of EVs from low volume of plasma, potentially improving disease diagnosis and biomarker discovery.(c) 2022 Elsevier B.V. All rights reserved.

Automated summary

Extracellular vesicles (EVs) released from tumor cells and tissues are involved in intercellular communication and regulation of biological processes. The study introduces a TiO2-mediated multiomics EV preparation method (TiMMEP) for isolating and profiling proteins and metabolites from a small volume of plasma. By capturing EVs using TiO2 and releasing proteins and metabolites with acetonitrile, the method enables high-throughput analysis of EVs, potentially improving disease diagnosis and biomarker discovery.