Objective: To establish a method of negative selection purification-enrichment (NSPE) extract exosomes from serum efficiently and rapidly, and exosomes were identified. Methods: A large amount of low density lipoproteins (LDL) interference in human serum was removed based on the principle of combining the low density lipoprotein receptor with low density lipoprotein. Then, exosomes were enriched by the principle of bidentate binding between the phosphate groups of phospholipid molecules on the surface of exosomes and ZrO2. Nanoflow cytometry and transmission electron microscopy were used to detect the particle size distribution, particle concentration and morphological characteristics of exosomes, and western blotting was used to identify surface specific proteins expression of exosomes. MicroRNAs (miRNAs) in exosomes were analyzed for gene pathway using the Gene Ontology database and the Kyoto Encyclopedia of Genes and Genomes database. Results: The exosomes in serum were effectively extracted using the method here within 60 minutes. The concentration of exosome particles was about (2.94±0.3)×1010 particles·mL-1, and the particle size was (88.07±2.94) nm. The expressions of exosome surface specific proteins, TSG101+ and CD9+, were detected. MiRNAs derived from exosomes were related to immune process and metabolic process. Conclusion: The established NSPE method could be used for rapid and efficient extraction of exosomes from serum for health monitoring and medical examination.
LI Xue-jie, FU Jie, JI Yuan-kai, SONG Hai-feng
. Extraction method and identification of serum exosomes*[J]. Chinese Journal of Pharmaceutical Analysis, 2022
, 42(8)
: 1400
-1406
.
DOI: 10.16155/j.0254-1793.2022.08.13
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