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Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry
Published on: October 24, 2018
Top-down proteomic analysis by MALDI-TOF profiling: Concentration-independent biomarkers
Gary L Nelsestuen1, Stephen B Harvey, Yan Zhang
1Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, MN, USA. nelse002@umn.edu.
Proteomics. Clinical Applications
|December 8, 2010
Summary
Analyzing protein isoform ratios offers a novel biomarker approach. Stable individual ratios, unlike total protein changes, may significantly enhance disease diagnosis sensitivity through longitudinal monitoring.
Area of Science:
- Biochemistry
- Proteomics
- Clinical Diagnostics
Background:
- Numerous protein biomarkers correlate with advanced disease, yet clinical assay development is limited.
- Current biomarker strategies often target total protein concentration changes, similar to acute phase reactants.
- Novel approaches are needed to identify sensitive and specific disease indicators.
Purpose of the Study:
- To explore the utility of intact protein isoform ratios as novel biomarkers.
- To investigate if stable, individual-specific protein isoform patterns can improve diagnostic sensitivity.
- To assess the potential of analyzing changes within protein isoforms rather than total protein levels.
Main Methods:
- Review of approaches focusing on intact protein isoform ratios, including proteolysis and glycoisoform changes.
- Application of an inexpensive method for large sample group analysis to identify protein polymorphisms.
- Longitudinal analysis of individual protein isoform stability.
Main Results:
- Identified protein polymorphisms, including the first structural polymorphism of apolipoprotein C1, linked to obesity.
- Observed altered isoform distributions, C- and N-terminal proteolysis, glycoisoform ratio changes, and sulfhydryl oxidation.
- Found protein isoform ratios to be highly stable within individuals, suggesting potential for increased diagnostic sensitivity.
Conclusions:
- Intact protein isoform ratios offer a biomarker strategy distinct from total protein concentration changes.
- Longitudinal analysis of stable individual protein isoform ratios may increase biomarker sensitivity significantly.
- Protein changes exceeding healthy ranges might be less common than alterations in isoform ratios.
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