A novel strategy for biomarker discovery: Neutral loss-dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptides
Circulating extracellular vehicles (EVs) play a crucial role in intercellular communication by transporting a diverse array of biological signaling molecules. Growing evidence indicates that phosphoproteins within these EVs hold promise as biomarkers for diagnostic and prognostic assessments, partic...
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Elsevier
2025-08-01
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| Series: | Talanta Open |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666831925000025 |
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| author | Dong Wei Hao Zhang Mingyang Li Liuxi Chu |
| author_facet | Dong Wei Hao Zhang Mingyang Li Liuxi Chu |
| author_sort | Dong Wei |
| collection | DOAJ |
| description | Circulating extracellular vehicles (EVs) play a crucial role in intercellular communication by transporting a diverse array of biological signaling molecules. Growing evidence indicates that phosphoproteins within these EVs hold promise as biomarkers for diagnostic and prognostic assessments, particularly in oncology. In this study, EVtrap (extracellular vesicle total recovery and purification) was utilized to isolate urinary EVs. We then combined this with an advanced liquid chromatography-tandem mass spectrometry (LC−MS/MS) technique to perform neutral loss-information-dependent acquisition-enhanced product ion (NL-IDA-EPI) analysis. This technique is specifically tailored to quantify endogenous phosphopeptides in urinary EVs by monitoring a consistent mass loss of 49 Da (H3PO4, doubly charged state) in the MS2 spectra. In this study, over 2,000 putative phosphopeptide signals were detected in a 10-mL urine sample without the need for prior phosphopeptide enrichment, thus simplifying the process and potentially minimizing sample bias. Comparative analysis of samples from high- and low-risk prostate cancer patients and a healthy control group revealed six differentially expressed phosphopeptides. These phosphopeptides serve as a promising panel of potential biomarkers, highlighting the effectiveness of our analytical framework in tracking cancer-related signaling events via phosphorylation within EVs using NL-IDA-EPI. |
| format | Article |
| id | doaj-art-31d8253f17d64603a4367090aee19219 |
| institution | Kabale University |
| issn | 2666-8319 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Talanta Open |
| spelling | doaj-art-31d8253f17d64603a4367090aee192192025-01-04T04:57:15ZengElsevierTalanta Open2666-83192025-08-0111100399A novel strategy for biomarker discovery: Neutral loss-dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptidesDong Wei0Hao Zhang1Mingyang Li2Liuxi Chu3School of Life & Health Sciences, Fuyao University of Science and Technology, Fuzhou, Fujian 350116, China; State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, Jiangsu, ChinaSchool of Life & Health Sciences, Fuyao University of Science and Technology, Fuzhou, Fujian 350116, China; EVLiXiR Biotech, Nanjing 210032, China; Corresponding author at: School of Life & Health Sciences, Fuyao University of Science and Technology, Fuzhou, Fujian 350116, China.State Key Laboratory of Digital Medical Engineering, School of Biological Science and Medical Engineering, Southeast University, Nanjing 210096, Jiangsu, ChinaAffiliated Cixi Hospital, Wenzhou Medical University, Ningbo, Zhejiang 315300, China; State Key Laboratory of Macromolecular Drugs and Large-scale Preparation, School of Pharmaceutical Science, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China; Corresponding author at: Affiliated Cixi Hospital, Wenzhou Medical University, Ningbo, Zhejiang 315300, China.Circulating extracellular vehicles (EVs) play a crucial role in intercellular communication by transporting a diverse array of biological signaling molecules. Growing evidence indicates that phosphoproteins within these EVs hold promise as biomarkers for diagnostic and prognostic assessments, particularly in oncology. In this study, EVtrap (extracellular vesicle total recovery and purification) was utilized to isolate urinary EVs. We then combined this with an advanced liquid chromatography-tandem mass spectrometry (LC−MS/MS) technique to perform neutral loss-information-dependent acquisition-enhanced product ion (NL-IDA-EPI) analysis. This technique is specifically tailored to quantify endogenous phosphopeptides in urinary EVs by monitoring a consistent mass loss of 49 Da (H3PO4, doubly charged state) in the MS2 spectra. In this study, over 2,000 putative phosphopeptide signals were detected in a 10-mL urine sample without the need for prior phosphopeptide enrichment, thus simplifying the process and potentially minimizing sample bias. Comparative analysis of samples from high- and low-risk prostate cancer patients and a healthy control group revealed six differentially expressed phosphopeptides. These phosphopeptides serve as a promising panel of potential biomarkers, highlighting the effectiveness of our analytical framework in tracking cancer-related signaling events via phosphorylation within EVs using NL-IDA-EPI.http://www.sciencedirect.com/science/article/pii/S2666831925000025Extracellular VesiclesNeutral lossPhosphopeptideNL-IDA-EPIProstate cancer |
| spellingShingle | Dong Wei Hao Zhang Mingyang Li Liuxi Chu A novel strategy for biomarker discovery: Neutral loss-dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptides Talanta Open Extracellular Vesicles Neutral loss Phosphopeptide NL-IDA-EPI Prostate cancer |
| title | A novel strategy for biomarker discovery: Neutral loss-dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptides |
| title_full | A novel strategy for biomarker discovery: Neutral loss-dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptides |
| title_fullStr | A novel strategy for biomarker discovery: Neutral loss-dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptides |
| title_full_unstemmed | A novel strategy for biomarker discovery: Neutral loss-dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptides |
| title_short | A novel strategy for biomarker discovery: Neutral loss-dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptides |
| title_sort | novel strategy for biomarker discovery neutral loss dependent acquisition of circulating extracellular vesicles for the detection of phosphopeptides |
| topic | Extracellular Vesicles Neutral loss Phosphopeptide NL-IDA-EPI Prostate cancer |
| url | http://www.sciencedirect.com/science/article/pii/S2666831925000025 |
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