| Title |
Comparison of commercial exosome isolation kits for circulating exosomal microRNA profiling
|
|---|---|
| Published in |
Analytical & Bioanalytical Chemistry, April 2018
|
| DOI | 10.1007/s00216-018-1052-4 |
| Pubmed ID | |
| Authors |
Meng Ding, Cheng Wang, Xiaolan Lu, Cuiping Zhang, Zhen Zhou, Xi Chen, Chen-Yu Zhang, Ke Zen, Chunni Zhang |
| Abstract |
Circulating exosomal microRNAs (miRNAs) are valuable biomarker candidates; however, information on the characterization and mutual agreement of commercial kits for circulating exosomal miRNA profiling is scarce. Here, we analyzed the advantages and weaknesses of four commonly used commercial kits for exosomal miRNA profiling and their application to the sample of serum and/or plasma, respectively. NanoSight and Western blotting were conducted to evaluate the efficiency and purity of the isolated exosomes. In our conditions, the size distribution of the isolated particles was appropriate (40-150 nm), and ExoQuick™ Exosome Precipitation Solution (EXQ) generated a relatively high yield of exosomes. Nevertheless, albumin impurity was ubiquitous for all the four kits, and Total Exosome Isolation for serum or plasma (TEI) yielded a relatively pure isolation. We further performed Illumina sequencing combined with RT-qPCR to determine the ability of these kits for miRNA profiling. There was significant correlation of the exosomal miRNA profile and specific miRNAs between kits, but with differences depending on methods. exoRNeasy Serum/Plasma Midi Kit (EXR) and EXQ performed better in the specific exosomal miRNAs recovery. Intraassay CVs for specific miRNA measurement were 0.88-3.82, 1.19-3.77, 0-2.70, and 1.23-9.11% for EXR, TEI, EXQ, and RIBO™ Exosome Isolation Reagent (REI), respectively. In each kit, serum yielded a higher abundance of exosomes and exosomal miRNAs than plasma, yet with more albumin impurity. In conclusion, our data provide some valuable guidance for the methodology of disease biomarker identification of circulation exosomal miRNAs. Graphical abstract Circulating exosomal microRNAs (miRNAs) are valuable biomarker candidates; however, information on the characterization and mutual agreement of commercial kits for circulating exosomal miRNA profiling is scarce. In this study, we compared four commonly used commercially available kits for exosomal miRNAsextraction and analyzed the advantages and weaknesses of each kit and their application to the sample ofserum and/or plasma. |
X Demographics
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Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Mexico | 1 | 50% |
| Unknown | 1 | 50% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Scientists | 2 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 143 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 143 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 22 | 15% |
| Researcher | 20 | 14% |
| Student > Master | 18 | 13% |
| Student > Bachelor | 11 | 8% |
| Student > Doctoral Student | 10 | 7% |
| Other | 22 | 15% |
| Unknown | 40 | 28% |
| Readers by discipline | Count | As % |
|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 32 | 22% |
| Medicine and Dentistry | 18 | 13% |
| Agricultural and Biological Sciences | 16 | 11% |
| Engineering | 4 | 3% |
| Pharmacology, Toxicology and Pharmaceutical Science | 4 | 3% |
| Other | 17 | 12% |
| Unknown | 52 | 36% |
Attention Score in Context
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