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Isolation and Analysis of Traceable and Functionalized Extracellular Vesicles from the Plasma and Solid Tissues
Published on: October 17, 2022
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Defining the Soluble and Extracellular Vesicle Protein Compartments of Plasma Using In-Depth Mass Spectrometry-Based
Nidhi Sharma1,2, Silvia Angori1, AnnSofi Sandberg1,2
1Department of Oncology-Pathology, Karolinska Institute, 171 77 Stockholm, Sweden.
Journal of Proteome Research
|August 14, 2024
Summary
Plasma-derived extracellular vesicles (pEVs) hold promise as disease biomarkers. A new workflow using size exclusion chromatography and mass spectrometry efficiently identifies pEV proteins from minimal plasma samples, aiding biomarker discovery in cancer patients.
Area of Science:
- Proteomics
- Extracellular Vesicle Biology
- Biomarker Discovery
Background:
- Plasma-derived extracellular vesicles (pEVs) contain potential disease biomarker proteins.
- Characterizing the pEV proteome is difficult due to low protein abundance and enrichment challenges.
Purpose of the Study:
- To develop and validate a streamlined workflow for comprehensive pEV proteome analysis from minimal plasma input.
- To identify potential protein biomarkers for metastatic lung adenocarcinoma (LUAD) and malignant melanoma (MM) using pEVs.
Main Methods:
- Size exclusion chromatography (SEC) combined with HiRIEF prefractionation and mass spectrometry (MS) was employed.
- Workflow performance was compared to long gradient MS using control pEVs.
- The SEC-HiRIEF-MS workflow was applied to plasma and pEVs from LUAD and MM patients.
Main Results:
- HiRIEF identified 2076 proteins in control pEVs, outperforming long gradient MS.
- The SEC-HiRIEF-MS workflow quantified 1583 proteins in cancer pEVs and 1468 proteins in plasma.
- Differential analysis identified 30 differentially expressed proteins (DEPs) between LUAD and MM patient groups.
Conclusions:
- The developed SEC-HiRIEF-MS workflow enables comprehensive pEV proteomics with minimal sample input.
- pEVs contain unique EV markers and cancer-related proteins, distinct from the plasma proteome.
- This approach shows significant potential for EV biomarker discovery in cancer.
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