Proteomics comparison of exosomes from serum and plasma between ultracentrifugation and polymer‐based precipitation kit methods

Abstract

Exosomes are vesicles with sizes ranging from 30 to 150 nm. The analysis and detection of blood exosomes offers an effective route for cancer diagnosis, prognosis assessment, and therapeutic evaluation of diseases. Due to the difference in separation procedure, collection method and the usage of anticoagulants, serum and plasma samples show diversity test results. In order to evaluate the isolation effect of exosomes in serum and plasma samples, two commonly used exosomal isolation methods, ultracentrifugation and polymer‐based precipitation kit, were used, respectively. And the isolation effects were evaluated by comparing the composition and abundant of proteins from isolated exosomes based on MS‐based proteomics analysis. The results showed that the plasma exosomes extracted by ultracentrifugation identified more exosome biomarkers, and the concentrations of these biomarkers were higher than others. And plasma exosomes could be a better sample for blood‐based proteomics research of exosomes. It would be more useful for future targeted biomarker discovery.

An insight into small extracellular vesicles: Their roles in colorectal cancer progression and potential clinical applications

“…145 Furthermore, there is still a debate whether plasma or serum is a better source of sEVs for clinical applications. Though plasma sEVs are deemed to contain more sEVs biomarkers and better for diagnosis than serum sEVs, 146 plasma contains platelets that are able to release several types of membrane vesicles, 147 which will confound the measurement of circulating miRNA biomarkers in plasma, 148,149 and even more nonvesicle miRNAs. 150 Nevertheless, compared to plasma, sEVs originated miRNAs are more highly expressed in serum sEVs samples.…”

Comparative Evaluation of Methods for Isolating Small Extracellular Vesicles Derived from Pancreatic Cells

“…However, our study results demonstrated that DGUC and IAC methods produced sEVs with better purity than UC. Besides, the interpretation of results should be cautious when commercial sEVs isolation kits based on the Co-P method was used 38,45 . The combination of several isolation methods may produce purer sEVs.…”

Circulating Exosomal miRNA as Diagnostic Biomarkers of Neurodegenerative Diseases

“…To facilitate the clinical application of exosomes, efficient separation and storage methods are essential. The current methods for isolating exosomes primarily comprise ultracentrifugation (Baranyai et al, 2015), the microfluidic chip method (Zhang et al, 2019), the antibody affinity capture method (Mathivanan et al, 2010), the polymer precipitation method (Cao F. et al, 2019), and gel exclusion chromatography (Hayashi et al, 2019). Of these, the most commonly used method is ultracentrifugation.…”