Proteomic toolbox to standardize the separation of extracellular vesicles and lipoprotein particles

Published

September 7, 2018

Author(s)

Tingting Wang, Illarion Turko

Abstract

Circulating in blood, extracellular vesicles (EVs) and lipoprotein particles (LPs) have diagnostic and prognostic value. To unambiguously define their functions, separation protocols need to be developed. However, because of their similar size and density, traditional approaches to separate EVs and LPs often fail to provide the required resolution. Further development and standardization of affinity-based protocols is necessary and a quantitative method is needed to assess the efficiency of LPs depletion from EVs samples. In the present study, we propose the simultaneous quantification of three groups of proteins by mass spectrometry as a toolbox to evaluate prospective separation protocols. We generated 15N- labeled internal standards for quantification of (i) EVs-specific proteins, (ii) all classes and subclasses of apolipoproteins constituting LPs, and (iii) major serum proteins. These standards were then used to evaluate the performance of size exclusion chromatography, heparin- Sepharose, lipopolysaccharide-Sepharose, (2-hydroxypropyl)-β-cyclodextrin-Sepharose, and concanavalin A-Sepharose in separating serum EVs and LPs.

Citation

ACS Journal of Proteome Research

Volume

17

Issue

9

Pub Type

Journals

Download Paper

DOI Link

Keywords

extracellular vesicles, lipoproteins, apolipoproteins, separation, mass spectrometry, QconCATs

Citation

Wang, T. and Turko, I. (2018), Proteomic toolbox to standardize the separation of extracellular vesicles and lipoprotein particles, ACS Journal of Proteome Research, [online], https://doi.org/10.1021/acs.jproteome.8b00225 (Accessed December 13, 2024)