Journal
CYTOMETRY PART A
Volume 93A, Issue 12, Pages 1207-1212Publisher
WILEY
DOI: 10.1002/cyto.a.23641
Keywords
biomarker; centrifugation; circulating cell-free DNA; circulating tumor cells; exosomes; extracellular vesicles; flow cytometry; microparticles; miRNA; tumor-educated platelets
Categories
Funding
- Netherlands Organisation for Scientific Research-Domain Applied and Engineering Sciences (NWO-TTW) [VENI 15924, 14198, VENI 13681]
Ask authors/readers for more resources
Biomarkers in the blood of cancer patients include circulating tumor cells (CTCs), tumor-educated platelets (TEPs), tumor-derived extracellular vesicles (tdEVs), EV-associated miRNA (EV-miRNA), and circulating cell-free DNA (ccfDNA). Because the size and density of biomarkers differ, blood is centrifuged to isolate or concentrate the biomarker of interest. Here, we applied a model to estimate the effect of centrifugation on the purity of a biomarker according to published protocols. The model is based on the Stokes equation and was validated using polystyrene beads in buffer and plasma. Next, the model was applied to predict the biomarker behavior during centrifugation. The result was expressed as the recovery of CTCs, TEPs, tdEVs in three size ranges (1-8, 0.2-1, and 0.05-0.2 mu m), EV-miRNA, and ccfDNA. Bead recovery was predicted with errors <18%. Most notable cofounders are the 22% contamination of 1-8 mu m tdEVs for TEPs and the 8-82% contamination of <1 mu m tdEVs for ccfDNA. A Stokes model can predict biomarker behavior in blood. None of the evaluated protocols produces a pure biomarker. Thus, care should be taken in the interpretation of obtained results, as, for example, results from TEPs may originate from co-isolated large tdEVs and ccfDNA may originate from DNA enclosed in <1 mu m tdEVs. (c) 2018 The Authors. Cytometry Part A published by Wiley Periodicals, Inc. on behalf of International Society for Advancement of Cytometry.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available