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Proteomic Profile of EPS-Urine through FASP Digestion and Data-Independent Analysis
Published on: May 8, 2021
Assessing human urinary proteome using a mass spectrometry-based profiling system combined with magnetic
Yu-Chang Tyan1, Ming-Hui Yang, Tze-Wen Chung
1Department of Medical Imaging and Radiological Sciences, Kaohsiung Medical University, Kaohsiung, Taiwan. yctyan@kmu.edu.tw
Clinica Chimica Acta; International Journal of Clinical Chemistry
|October 20, 2012
Summary
This study developed a novel cleanup method using magnetic nanoparticles (MNPs) for improved urinary proteome analysis. The technique simplifies sample preparation, enabling better identification of proteins for diagnostic biomarker discovery.
Area of Science:
- Biochemistry
- Proteomics
- Analytical Chemistry
Background:
- Body fluid samples contain contaminants hindering mass spectrometry analysis.
- Urine is an ideal bio-sample rich in potential diagnostic biomarkers.
- Direct analysis of urine by mass spectrometry is often precluded by sample complexity.
Purpose of the Study:
- To establish optimal techniques for developing a urinary proteomic map.
- To propose a simplified sample preparation method for urine proteome analysis.
- To enhance the understanding of the urinary proteome.
Main Methods:
- Urine proteins were prepared using magnetic nanoparticles (MNPs) and a MACS separation column system.
- Protein identification was performed using reverse-phase nano-high-performance liquid chromatography-electrospray ionization tandem mass spectrometry (RP-nano-HPLC-ESI-MS/MS).
- Peptide fragmentation patterns were analyzed for protein identification.
Main Results:
- The cleanup approach demonstrated improved protein identification, exemplified by bovine serum albumin (BSA) in artificial urine.
- A total of 542 peptides, corresponding to 282 unique proteins, were identified from human urine samples.
- Fifty-four proteins were identified with higher confidence levels, with potential age-related increases in immunoproteins noted.
Conclusions:
- Optimal techniques were established for developing a proteomic map of urinary proteins.
- A cleanup method was proposed that significantly simplifies the urine sample preparation process.
- The developed method enhances the feasibility of urinary proteome analysis for biomarker discovery.
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