REAL

Characterization of extracellular vesicles by IR spectroscopy: fast and simple classification based on amide and C-H stretching vibrations

Mihály, Judith and Deák, Róbert and Szigyártó, Imola Csilla and Bóta, Attila and Beke-Somfai, T. and Varga, Zoltán (2016) Characterization of extracellular vesicles by IR spectroscopy: fast and simple classification based on amide and C-H stretching vibrations. BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES. ISSN 0005-2736 (In Press)

[img]
Preview
Text
BBA_11_1128.pdf - Accepted Version

Download (1MB) | Preview

Abstract

Extracellular vesicles isolated by differential centrifugation from Jurkat T-cell line were investigated by attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR). Amide and C-H stretching band intensity ratios calculated from IR bands, characteristic of protein and lipid components, proved to be distinctive for the different extracellular vesicle subpopulations. This proposed ’spectroscopic protein-to-lipid ratio’, combined with the outlined spectrum-analysis protocol is valid also for low sample concentrations (0.15-0.05 mg/ml total protein content) and can carry information about the presence of other non-vesicular formations such as aggregated proteins, lipoproteins and immune complexes. Detailed analysis of IR data reveals compositional changes of extracellular vesicles subpopulations: second derivative spectra suggest changes in protein composition from parent cell towards exosomes favoring proteins with -turns and unordered motifs at the expense of intermolecular -sheet structures. The IR-based protein-to-lipid assessment protocol was tested also for red blood cell derived microvesicles for which similar values were obtained. The potential applicability of this technique for fast and efficient characterization of vesicular components is high as the investigated samples require no further preparations and all the different molecular species can be determined in the same sample. The results indicate that ATR-FTIR measurements provide a simple and reproducible method for the screening of extracellular vesicle preparations. It is hoped that this sophisticated technique will have further impact in extracellular vesicle research.

Item Type: Article
Subjects: Q Science / természettudomány > QD Chemistry / kémia > QD01 Analytical chemistry / analitikai kémia
Depositing User: Judith Mihály
Date Deposited: 29 Sep 2017 18:20
Last Modified: 05 Apr 2023 06:44
URI: http://real.mtak.hu/id/eprint/64448

Actions (login required)

Edit Item Edit Item