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Licensed Unlicensed Requires Authentication Published by De Gruyter April 16, 2019

Comparison of six commercial serum exosome isolation methods suitable for clinical laboratories. Effect in cytokine analysis

  • Mónica Macías , Vera Rebmann , Beatriz Mateos , Nerea Varo , Jose Luis Perez-Gracia , Estibaliz Alegre and Álvaro González EMAIL logo

Abstract

Background

Exosomes are nanovesicles released by cells that can be detected in blood. Exosomes contain several molecules, such as cytokines that have potential utility as disease biomarkers. The aim of the present work is to compare six different commercial kits suitable for the clinical laboratory in relation to the efficiency and purity of exosome isolation, and their effect in subsequent cytokines analysis.

Methods

Serum exosomes were obtained from 10 volunteers using six commercial kits: exoEasy, ExoQuick, Exo-spin, ME kit, ExoQuick Plus and Exo-Flow. Exosome concentrations and size distributions were quantified by nanoparticle tracking analysis. Exosome markers CD63, CD9 and TSG101 were determined by Western blot. ApoB and albumin were measured using nephelometry. S100A9, CXCL5 and CXCL12 were measured using a Luminex assay.

Results

The concentration of particles obtained between different kits varied by a factor of 100. There was no correlation in particle concentrations extracted between different kits, except between ExoQuick and Exo-Flow. The highest exosome purity was achieved with ExoQuick Plus and exoEasy, while the lowest were achieved with ME and ExoQuick. Albumin was present in all exosome extracts analyzed and ApoB in all except those extracted with Exo-Flow and ME. Cytokine detection varied depending on the purification kit used and there was no correlation in cytokine concentrations between samples obtained with different kits.

Conclusions

Both the sample and the type of commercial kit used affect the efficiency and purity of exosome isolation. In addition, the exosome purification method deeply affects the capability to detect and quantify cytokines.


Corresponding author: Álvaro González, PhD, Department of Biochemistry, Clínica Universidad de Navarra, Avenida Pío XII 36, 31008 Pamplona, Spain; and Navarra Institute for Health Research (IdiSNA), Pamplona, Spain, Phone: +34 948 255400, Fax: +34 948 296 500
aEstibaliz Alegre and Álvaro González contributed equally to the work.
  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: This work was supported by “Fondo de Investigación Sanitaria” grant [PI14/00274]. We like to thank Dra. María Romero and Teresa Sendino for their support in the preparation of the manuscript.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2018-1297).


Received: 2018-12-05
Accepted: 2019-03-23
Published Online: 2019-04-16
Published in Print: 2019-09-25

©2019 Walter de Gruyter GmbH, Berlin/Boston

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