期刊
JOURNAL OF PROTEOMICS
卷 249, 期 -, 页码 -出版社
ELSEVIER
DOI: 10.1016/j.jprot.2021.104359
关键词
Extracellular vesicles; Targeted proteomics; QconCATs; Multiple reaction monitoring
Absolute quantification with mass spectrometry and isotope labeled internal standards has been utilized to develop and evaluate a new affinity-based approach to isolate extracellular vesicles (EVs) from human plasma. Peptides with affinity to EVs selected from phage library screening have been demonstrated as valuable ligands for EVs isolation, showing potential for drug delivery with no off-target side effects. Quantification of EVs proteins through MRM can also assist in identifying subpopulation specific markers for target-specific isolation.
Absolute quantification with mass spectrometry and isotope labeled internal standards has found broad applications in biomedical research. In the present research, it was used for developing and evaluating a new affinity based approach to isolate extracellular vesicles (EVs) from human plasma. First, a phage display peptide library was screened against EVs as a bait and absolute quantification of multiple proteins helped to select the best bait available. Then, absolute quantification was used to evaluate the efficiency of affinity chromatography on peptide-Sepharose. In summary, we have demonstrated that peptides with affinity to EVs selected from phage library screening can be valuable ligands for EVs isolation. Significance: Extracellular vesicles (EVs) have an important role in intercellular communication for all cell types. This makes EVs a promising new type of therapeutics capable to deliver drugs to specific sites with no off-target side effects. However, their isolation, and correct assignment of their biological function and properties remains an obscure field of research. In this study, we proposed to use MRM quantitation of a pattern of EVs and non-EVs proteins to develop a purification protocol based on affinity peptides selected from phage library screening. MRM quantification of EVs proteins can also help in identifying those that are subpopulation specific markers for further target-specific isolation.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据