Proteomic Analysis of Exosomes

The proteomic analysis of exosomes was performed using a label-free analysis technology. Protein was extracted from exosomes, quantified with BCA assay, digested by trypsin, and dried by centrifugal concentration. Then, the peptides were desalted using MonoSpin C18 desalting column (GL Sciences Inc., Japan, 5010-21701), analyzed by Orbitrap Fusion Lumos tandem mass spectrometry (Thermo Fisher Scientific) coupled to an EASY-nLC 1200 liquid chromatography system (Thermo Fisher Scientific). Protein identification and quantification were performed using Proteome Discoverer 2.4 (Database: Swissprot; taxonomy: Homo sapiens) with the default setting. The experiments and data analysis were offered by Wayen Biotechnologies (Shanghai, China). We used the Metascape platform online analytical tool to further analyze part of the proteomic analysis data.^{37 (link)} The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE^{38 (link)} partner repository with the dataset identifier PXD033839.

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Liu P., Yang S., Shao X., Li C., Wang Z., Dai H, & Wang C. (2024). Mesenchymal Stem Cells-Derived Exosomes Alleviate Acute Lung Injury by Inhibiting Alveolar Macrophage Pyroptosis. Stem Cells Translational Medicine, 13(4), 371-386.

Publication 2024

EASY-nLC 1200 liquid

Corresponding Organization : Harbin Medical University

Other organizations : Tianjin Third Central Hospital, Tianjin Medical University, Capital Medical University

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Variable analysis

independent variables

Proteomic analysis of exosomes

dependent variables

Protein identification and quantification

control variables

Protein extraction from exosomes
Protein quantification using BCA assay
Trypsin digestion of proteins
Desalting of peptides using MonoSpin C18 column
Analysis by Orbitrap Fusion Lumos tandem mass spectrometry
Protein identification and quantification using Proteome Discoverer 2.4 (Database: Swissprot; taxonomy: Homo sapiens)

positive controls

None specified

negative controls

None specified

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