STEP 01
Isolate total vesicles
Plasma or serum is processed with an EV isolation kit, following the laboratory's validated procedures, to obtain a total extracellular-vesicle fraction.
Methodology
We measure protein and microRNA biomarkers carried by brain-derived extracellular vesicles (BDEVs), isolated from plasma or serum through a standardised, quality-controlled workflow.
One biofluid sample
Such as blood (plasma or serum) — a minimally invasive starting point.
Two biomarker classes
Proteins and microRNA, measured from the same enriched fraction.
Every well in duplicate
Calibrated, controlled and tracked by analytical batch.
The workflow
Each stage is designed so that samples are compared on equal footing, and every measurement can be traced back to its plate and batch.
STEP 01
Plasma or serum is processed with an EV isolation kit, following the laboratory's validated procedures, to obtain a total extracellular-vesicle fraction.
STEP 02
The total fraction is selectively enriched for brain-derived vesicles, separating this population from the many other EVs circulating in biofluids (e.g. blood, urine, saliva…).
STEP 03
Nanoparticle tracking analysis (NanoSight) measures particle size and concentration. Every sample is then adjusted to a common particle input before any biomarker is measured.
STEP 04
Target proteins are quantified with target-specific ELISAs, each calibrated against a recombinant-protein standard curve and run with positive, negative and total-EV reference controls.
STEP 05
Each target microRNA binds a complementary probe captured on the plate, forming an RNA–DNA duplex that a duplex-selective detection system turns into a colorimetric signal.
STEP 06
Both pathways are read on a common microplate platform (Tecan), every sample in technical duplicate, with each plate and analytical batch recorded for quality review and statistics.
Calibration and controls
microRNA assays are quantified against a dilution series of synthetic microRNA for each target, processed through the full hybridisation and detection steps.
Around it, a panel of controls separates true signal from background and checks that only the intended sequence is detected.
| Control / reference | What it tells us |
|---|---|
| Synthetic target microRNA series | Target-specific quantitative calibration |
| Complementary target + probe | Positive hybridisation and detection control |
| Probe without target | Background from the probe and detection chemistry |
| Non-complementary RNA + probe | Hybridisation specificity |
| Target without probe | Background without probe capture |
| Detection-system reference | Monitors downstream detection performance |
Scientific honesty
Brain origin comes from the enrichment strategy. Particle analysis characterises the fraction but does not, on its own, prove where vesicles come from.
Equalising particle input reduces variability between samples. It does not establish purity or remove batch effects by itself.
Batch correction is assessed against how clinical groups are distributed across plates — statistics cannot undo complete confounding.
Further validated assay specifications and performance characteristics can be shared with collaborators where needed for reproducibility, while protecting our proprietary technology.