Purification and Phosphoproteomic Analysis of Plasma-Derived Extracellular Vesicles

A successful phosphoproteomics analysis of extracellular vesicles (EVs) requires a unique approach, fine-tuned to address the challenges that have plagued plasma-based biomarker discovery. Here, I detail a procedure, which combines EVtrap-based high-recovery EV isolation, phase-transfer surfactant m...

Full description

Saved in:
Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2504; p. 147
Main Author Iliuk, Anton B
Format Journal Article
LanguageEnglish
Published United States 2022
Subjects
Biomarkers - metabolism
Extracellular Vesicles - metabolism
Phosphopeptides - metabolism
Plasma - chemistry
Proteome - metabolism
Extracellular vesicles (EVs)
Plasma
Liquid biopsy
Phosphoproteomics
Exosomes
EVtrap
Online AccessGet more information

Cover

More Information
Summary:A successful phosphoproteomics analysis of extracellular vesicles (EVs) requires a unique approach, fine-tuned to address the challenges that have plagued plasma-based biomarker discovery. Here, I detail a procedure, which combines EVtrap-based high-recovery EV isolation, phase-transfer surfactant method for protein extraction, and PolyMAC-based enrichment of phosphopeptides. The combination of these methods provides a highly effective strategy for EV-based phosphoproteome analysis and leads to the discovery of novel phospho-markers previously undetectable.
ISSN:1940-6029
DOI:10.1007/978-1-0716-2341-1_11