Proteomics protocol for obtaining extracellular vesicle from human plasma using asymmetrical flow field-flow fractionation technology

Na Li; Yumeng Yan; Bowen Wu; Jifeng Wang; Fuquan Yang

doi:10.1016/j.xpro.2023.102515

Proteomics protocol for obtaining extracellular vesicle from human plasma using asymmetrical flow field-flow fractionation technology

STAR Protoc. 2023 Dec 15;4(4):102515. doi: 10.1016/j.xpro.2023.102515. Epub 2023 Sep 23.

Authors

Na Li¹, Yumeng Yan¹, Bowen Wu¹, Jifeng Wang¹, Fuquan Yang²

Affiliations

¹ Key Laboratory of Protein and Peptide Pharmaceuticals & Laboratory of Proteomics, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China.
² Key Laboratory of Protein and Peptide Pharmaceuticals & Laboratory of Proteomics, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China; University of Chinese Academy of Sciences, Beijing 100049, China. Electronic address: fqyang@ibp.ac.cn.

Abstract

Plasma extracellular vesicles (EVs) represent a potential resource for biomarkers of multiple diseases. Here, we present a protocol for obtaining EVs from human plasma using asymmetrical flow field-flow fractionation technology. We describe steps for using tandem mass tags to perform comparative proteomic studies of a large clinical cohort. We then detail targeted quantitative analysis of differential proteins based on a parallel reaction monitoring technique. For complete details on the use and execution of this protocol, please refer to Wu et al. (2020)¹ and Li et al. (2023).².

Keywords: Cell Biology; Cell Isolation; Cell Separation/Fractionation; Clinical Protocol; Health Sciences; Mass Spectrometry; Molecular/Chemical Probes; Protein Expression and Purification; Proteomics.

MeSH terms

Extracellular Vesicles*
Fractionation, Field Flow*
Humans
Proteomics
Technology