Automated EV proteomics: High-throughput SAX-based Isolation, on-bead lysis, and rapid digestion for reproducible LC-MS/MS analysis
How the technology works
- Electrostatic “Netting. ”Hyper-porous strong anion-exchange (SAX) magnetic beads selectively bind membrane-bound extracellular vesicles (EVs) while letting soluble high-abundance proteins flow through1.
- On-Bead Lysis & PAC. Captured vesicles are lysed, reduced (5 mM TCEP) and alkylated (10 mM CAA), then subjected to protein aggregation capture (PAC) directly on the same bead surface2.
- Organic Shrink-Wash. Dual 95% acetonitrile washes precipitate remaining contaminants and compress dynamic range.
- Rapid Digestion. A 2-hour, 47 °C trypsin (±Lys-C) digest releases peptides for LC-MS/MS; digestion CVs remain below 21% across replicates1.
- 96-Sample Automation. The KingFisher™-compatible protocol finishes in ≈3 h hands-off, with .bdz scripts supplied on request.
Validated by peer-reviewed studies: High-coverage EV proteomics enables robust biomarker discovery and diagnostic differentiation in neurodegenerative disease
| Study | Sample Set & Goal | Key Findings Using Mag-Net |
| Genome Sciences, Univ. of Washington (April 2024) | 40-patient pilot: Alzheimer’s disease dementia (ADD), Parkinson’s with and without dementia, and age-matched controls | - Single-run DIA quantified 4,163 protein groups per sample versus 1,088 from unfractionated plasma1. - 204 proteins separated ADD from all other groups (ROC AUC 0.98), indicating diagnostic potential1. |
| Methodology paper (bioRxiv 2023 / Nat. Commun. 2024) | Technical benchmarking across triplicate preps | - Routine detection of >37,000 peptides; 21.4% carried predicted transmembrane domains—an under-represented class in plasma studies. - Inter-prep Pearson r = 0.978–0.985; median protein CV ≈ 21%1. |
These data confirm that Mag-Net’s dual action—EV enrichment plus abundant-protein depletion—opens a proteomic window previously accessible only through extensive fractionation or multi-dimensional workflows.
Next-gen EV enrichment: Mag-Net HP enhances binding capacity, reduces Background, and enables Ultra-low input plasma proteomics
What's new?
- New bead cocktail with higher binding capacity and reduced non-specific carry-over.
- Optimised buffers that maintain EV integrity yet enhance on-bead protein aggregation.
- Ultra-low input compatibility: validated down to 5 µL of EDRN-protocol plasma; scales to 50 µL for platelet-poor plasma.
Scalable EV Proteomics: Mag-Net HP powers biomarker discovery in neurodegeneration, oncology, and large-scale plasma cohorts
- Neurodegeneration: Detection of CNS-derived EV markers (CD9, CD63, TSG101 enriched >30-fold) enables minimally invasive tracking of Alzheimer’s or Parkinson’s pathology1.
- Liquid Biopsy Discovery: High coverage of transmembrane and lipid-anchored proteins opens avenues for oncology and cardiovascular biomarker panels1.
- Population-Scale Cohorts: 96-well automation and consumable cost < USD 5 per sample position Mag-Net HP for epidemiological studies requiring thousands of samples.
Mag-Net HP performance specs: Ultra-low input, High-throughput EV Proteomics with deep coverage and global availability
| Parameter | Value |
| Minimum plasma input | 5 µL (EDRN SOP)2 |
| Throughput | 96 samples / 3 h (KingFisher Flex) |
| Proteome depth (single DIA run) | >4,000 proteins, >37,000 peptides1 |
| EV marker enrichment | >30× (CD9, CD63, TSG101)1 |
| Reproducibility | Median protein CV ≃ 21%; Pearson r > 0.9781 |
Availability The Mag-Net HP Kit (96 reactions) includes pre-dispensed magnetic beads, optimized buffers, and enzyme-ready reagents—streamlining EV proteomics from sample to analysis.
