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Citation: ZENG Qiong-Lan, WU Li, LI Shui-Ming, WANG Yong. Comparison of Separation Methods for Isolation in Analysis of Human Saliva-derived Exosomes Proteomics by Nano-liquid Chromatography-Mass Spectrometry[J]. Chinese Journal of Analytical Chemistry, ;2018, 46(9): 1408-1414. doi: 10.11895/j.issn.0253-3820.181261 shu

Comparison of Separation Methods for Isolation in Analysis of Human Saliva-derived Exosomes Proteomics by Nano-liquid Chromatography-Mass Spectrometry

1.
College of Life and Ocean Science, Shenzhen Key Laboratory of Marine Bioresources and Ecology, Shenzhen University, Shenzhen 518060, China;
2.
College of Life and Ocean Science, Shenzhen Key Laboratory of Microbial Genetic Engineering, Shenzhen 518060, China

Received Date: 19 April 2018
Revised Date: 19 July 2018

Fund Project: This work was supported by the Scientific Plan Program of Shenzhen, China (No. JCYJ20170818142154551)

Exosomes are a series of extracellular vesicles, which are the source of biomarkers. However, there are no sensitive and efficient methods for the isolation of saliva exosome proteins. In this work, we explored the effect of isolation of saliva exosome by ExoQuick (EQ) and ultracentrifugation (UC) method, and the effect of UREA buffer, RIPA lysate, and SDS lysate on exosome protein extraction by nano-liquidchromatography-high-resolution tandem mass spectrometry. The quantitative results of Brodford method showed that the protein content of exosomes obtained by EQ method from 0.5 mL of saliva was higher than that by UC method from 2 mL of saliva. Further mass spectrometry analysis showed that the former identified more proteins than the latter. The effect of the combination of using EQ method and UREA buffer to obtain saliva exosomes proteins was the best. With this method, a total of 194 proteins were identified. The method had good reproducibility, low sample requirement, and stable mass spectrometry detection, and could provide methodological support for finding markers of disease in saliva exosomes.
- Nano liquid chromatography-high resolution tandem mass spectrometry,
- Saliva-exosome,
- ExoQuick,
- Ultracentrifugation
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