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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
Published/Copyright: April 16, 2019
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 57 Issue 10

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.

Keywords: CXCL5; CXCL12; cytokines; exosomes; purification; S100A9
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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https://doi.org/10.1515/cclm-2018-1297
Keywords for this article
CXCL5; CXCL12; cytokines; exosomes; purification; S100A9

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Blood biomarkers in neurology: “a call to arms” for laboratory professionals
  4. Reviews
  5. Diagnostic accuracy of glycated hemoglobin for gestational diabetes mellitus: a systematic review and meta-analysis
  6. Laboratory medicine: health evaluation in elite athletes
  7. Prostate cancer screening: guidelines review and laboratory issues
  8. Opinion Papers
  9. Extra-analytical sources of uncertainty: which ones really matter?
  10. Benefits and harms of wellness initiatives
  11. Genetics and Molecular Diagnostics
  12. Analytical and clinical validation of a novel amplicon-based NGS assay for the evaluation of circulating tumor DNA in metastatic colorectal cancer patients
  13. General Clinical Chemistry and Laboratory Medicine
  14. Pre-analytical practices for routine coagulation tests in European laboratories. A collaborative study from the European Organisation for External Quality Assurance Providers in Laboratory Medicine (EQALM)
  15. Preanalytical robustness of blood collection tubes with RNA stabilizers
  16. Continual improvement of the pre-analytical process in a public health laboratory with quality indicators-based risk management
  17. Comparison of six commercial serum exosome isolation methods suitable for clinical laboratories. Effect in cytokine analysis
  18. A multicenter study to evaluate harmonization of assays for N-terminal propeptide of type I procollagen (PINP): a report from the IFCC-IOF Joint Committee for Bone Metabolism
  19. Correlations between serum and CSF pNfH levels in ALS, FTD and controls: a comparison of three analytical approaches
  20. Dynamics of extracellular matrix proteins in cerebrospinal fluid and serum and their relation to clinical outcome in human traumatic brain injury
  21. Free light chains in the cerebrospinal fluid. Comparison of different methods to determine intrathecal synthesis
  22. Reference Values and Biological Variations
  23. Reference interval by the indirect approach of serum thyrotropin (TSH) in a Mediterranean adult population and the association with age and gender
  24. Next-generation reference intervals for pediatric hematology
  25. Hematology and Coagulation
  26. Preliminary evaluation of a new flow cytometry method for the routine hematology workflow
  27. Diabetes
  28. Trueness assessment of HbA1c routine assays: are processed EQA materials up to the job?
  29. Infectious Diseases
  30. Utility of procalcitonin for differentiating cryptogenic organising pneumonia from community-acquired pneumonia
  31. A high C-reactive protein/procalcitonin ratio predicts Mycoplasma pneumoniae infection
  32. Letters to the Editor
  33. Evaluation of reference change values for a hs-cTnI immunoassay using both plasma samples of healthy subjects and patients and quality control samples
  34. Outlier removal methods for skewed data: impact on age-specific high-sensitive cardiac troponin T 99th percentiles
  35. Comparison of precision and operational performances across six immunochemistry analyzers
  36. Evaluation of the possible interference of abiraterone therapy on testosterone immunoassay
  37. Erroneous thyroid and steroid hormones profile due to anti-streptavidin antibodies
  38. Reference values for 24,25-dihydroxyvitamin D and the 25-hydroxyvitamin D/24,25-dihydroxyvitamin D ratio
  39. Pre-analytical error in a hematology laboratory: an avoidable cause of compromised quality in reporting
  40. Stability of tubular damage markers epidermal growth factor and heparin-binding EGF-like growth factor in urine
  41. Blood from heparin tubes is an acceptable alternative to assess hematocrit determination
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