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Updated: Sep 11, 2025
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A High Throughput, Multiplexed and Targeted Proteomic CSF Assay to Quantify Neurodegenerative Biomarkers and Apolipoprotein E Isoforms Status
Published on: October 20, 2016
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Luminal Cerebrovascular Proteomics
Sophia M Shi1,2,3,4, Carolyn R Bertozzi1,2,5, Tony Wyss-Coray2,3,4,6
1Department of Chemistry, Stanford University, Stanford, CA, USA.
Bio-Protocol
|August 13, 2025
Summary
Researchers developed a new in vivo proteomic method to map the brain
Area of Science:
- Neuroscience
- Proteomics
- Cell Biology
Background:
- Brain endothelial cells form the blood-brain barrier, crucial for central nervous system (CNS) homeostasis.
- Distinct luminal and abluminal membrane compositions mediate cerebrovascular functions.
- Current proteomic methods lack spatial resolution to differentiate these membrane compartments.
Purpose of the Study:
- To develop a high-resolution in vivo proteomic strategy for profiling the luminal cerebrovascular surface.
- To overcome limitations of existing techniques in discriminating between membrane compartments.
Main Methods:
- In vivo perfusion of a membrane-impermeable biotinylation reagent to label luminal surface proteins.
- Microvessel isolation and streptavidin-based enrichment of biotinylated proteins.
- Downstream mass spectrometry (LC-MS/MS) for protein identification.
Main Results:
- Successfully identified over 1,000 luminally localized proteins.
- Achieved significantly improved enrichment of canonical luminal markers compared to conventional methods.
- Generated a compartment-resolved atlas of the luminal cerebrovascular proteome.
Conclusions:
- The developed method provides a high-confidence, spatially resolved map of the luminal cerebrovascular proteome.
- Offers a scalable platform for studying endothelial surface biology in health and disease.
- Enables investigation of aging-related changes and identification of therapeutic targets.
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