Profiling early-stage breast cancer small extracellular vesicles using a DSPE-functionalised herringbone microfluidic chip
Aims
Current liquid biopsy platforms for detecting small extracellular vesicles (sEVs) lack sufficient sensitivity and specificity for early-stage breast cancer (BC) diagnosis. We aimed to develop and validate a microfluidic method to improve sEV capture and molecular profiling for clinical application.
Methods
We designed a herringbone microfluidic chip functionalised with DSPE-PEG for universal lipid-based sEV capture. The chip architecture enhances mixing and increases sEV-surface interactions under optimised flow conditions. sEVs were isolated from plasma of 30 early-stage BC patients and 10 healthy donors and profiled using multiplex fluorescent staining for EpCAM, PD-L1, Trop-2, and CD9.
Results
The platform achieved high sensitivity, detecting sEV concentrations down to 2×10⁷ particles/mL. sEV capture efficiency was significantly enhanced compared to straight channels. Molecular profiling revealed heterogeneous and stage-dependent expression of the three BC markers. Inter-group comparisons showed significant differences in marker expression between BC and healthy controls.
Conclusions
The DSPE-functionalised herringbone chip enables efficient, label-free sEV capture and multiplexed phenotyping of early-stage breast cancer in a rapid workflow. Its sensitivity and ability to differentiate patient-specific sEV signatures support its clinical potential for non-invasive early BC diagnosis and monitoring.