Titanium(IV) immobilized affinity chromatography facilitated phosphoproteomics analysis of salivary extracellular vesicles for lung cancer

Extracellular vesicles (EVs) play critical roles in intercellular communications, which contain valuable biomarkers for the detection of cancers. Phosphoproteomics analysis of human saliva EVs (sEVs) can help to discover lung cancer–related candidates. Due to the low abundance of phosphoproteins in...

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Published in:Analytical and bioanalytical chemistry 2022-05, Vol.414 (12), p.3697-3708
Main Authors: Wahid, Amir, Sohail, Amir, Wang, Huiyu, Guo, Miao, Zhang, Lu, Ji, Yin, Wang, Peng, Xiao, Hua
Format: Article
Language:English
Subjects:
Affinity
Affinity chromatography
Analytical Chemistry
Biochemistry
Biomarkers
Characterization and Evaluation of Materials
Chemistry
Chemistry and Materials Science
Chromatography
Extracellular vesicles
Food Science
Laboratory Medicine
Lung cancer
Methods
Monitoring/Environmental Analysis
Phosphoproteins
Physiological aspects
Proteomics
Research Paper
Saliva
Titanium
Titanium dioxide
Vesicles
Zirconium dioxide
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Summary:Extracellular vesicles (EVs) play critical roles in intercellular communications, which contain valuable biomarkers for the detection of cancers. Phosphoproteomics analysis of human saliva EVs (sEVs) can help to discover lung cancer–related candidates. Due to the low abundance of phosphoproteins in sEVs, an efficient, reproducible, and cost-effective strategy is required for their enrichment. Here, we compared the latest phosphopeptide techniques, including TiO 2 , ZrO 2 , CaTiO 3 , and Ti 4+ -IMAC (immobilized metal affinity chromatography) methods, for phosphopeptide isolation. Our data demonstrated that Ti 4+ -IMAC was the superior one. By using the optimized Ti 4+ -IMAC approach, we identified more than 500 sEV phosphopeptides. Quantitative proteomics was employed to comprehensively decipher the sEV phosphoproteome of the normal group ( n  = 6) and lung cancer group ( n  = 6). Accordingly, 524 and 333 phosphopeptides were enriched, respectively, which corresponded to 439 and 282 phosphoproteins. In total, 857 unique sEV phosphopeptides corresponding to 721 phosphoproteins were revealed. Among 493 identified phosphosites, 37 were upregulated (> 1.5) and 217 were downregulated (
ISSN:1618-2642
1618-2650
DOI:10.1007/s00216-022-04013-7