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A High Throughput, Multiplexed and Targeted Proteomic CSF Assay to Quantify Neurodegenerative Biomarkers and Apolipoprotein E Isoforms Status
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Targeted proteomic strategy for clinical biomarker discovery
Ralph Schiess1, Bernd Wollscheid, Ruedi Aebersold
1Institute of Molecular Systems Biology, ETH Zurich, Switzerland.
Molecular Oncology
|April 23, 2009
Summary
A new two-stage strategy enhances disease biomarker discovery in blood plasma. This method focuses on N-linked glycoproteins for sensitive, non-invasive, and quantitative disease detection.
Area of Science:
- Biochemistry
- Proteomics
- Biomarker Discovery
Background:
- Plasma proteome complexity and dynamic range hinder sensitive, high-throughput disease biomarker profiling.
- Existing methods struggle with large sample sets needed for reliable disease-indicating difference detection.
Purpose of the Study:
- To introduce a novel two-stage strategy for mass spectrometry (MS)-assisted discovery, verification, and validation of disease biomarkers.
- To overcome technological limitations in sensitive and high-throughput plasma proteome profiling.
Main Methods:
- A two-stage approach: 1. Discovery phase identifying N-linked glycoproteins with differential expression in tissues. 2. Verification/validation phase using targeted MS analysis (selected reaction monitoring - SRM) on plasma samples.
- Focus on N-glycoproteins, which are likely secreted or shed, to reduce complexity and target a relevant subproteome for remote disease sensing.
Main Results:
- The strategy enables robust identification of protein candidate panels.
- The workflow allows for selective monitoring of biomarkers in blood plasma with high sensitivity, reliability, and quantitative accuracy.
Conclusions:
- The N-glycoprotein-focused workflow provides a sensitive, non-invasive, and quantitative method for disease biomarker discovery and validation.
- This approach facilitates the reliable detection of disease-indicating differences through targeted analysis of tissue-derived glycoproteins in plasma.
