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Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular
Agnieszka Kamińska1, Magdalena E Marzec1, Ewa Ł Stępień1
1Department of Medical Physics, Marian Smoluchowski Institute of Physics, Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, 30-348 Kraków, Poland.
Molecules (Basel, Switzerland)
|August 27, 2021
Summary
This study optimized silicon surface functionalization for capturing urinary extracellular vesicles (uEVs). The optimal method used lactadherin protein at 25 µg/mL for efficient uEV binding.
Area of Science:
- Biomaterials Science
- Nanotechnology
- Biochemistry
Background:
- Urinary extracellular vesicles (uEVs) are crucial biomarkers for non-invasive disease diagnosis.
- Efficient capture and characterization of uEVs are essential for their clinical application.
- Current methods for uEV isolation and surface functionalization require optimization.
Purpose of the Study:
- To optimize silicon surface functionalization for enhanced urinary extracellular vesicle (uEV) capture.
- To investigate the impact of silane type and protein concentration on uEV binding efficiency.
- To characterize the functionalized biosensor surface and isolated uEVs.
Main Methods:
- Silicon surface functionalization using APTES and GOPS silanes.
- Immobilization of human lactadherin (LACT) protein for uEV capture.
- Surface characterization via ellipsometry, AFM, and ToF-SIMS.
- uEV isolation using filtration and ultracentrifugation.
- uEV characterization using TEM and tunable resistive pulse sensing.
Main Results:
- Gradual molecular layer growth observed on the silicon surface after functionalization steps.
- ToF-SIMS confirmed no significant changes in amino acid and lipid profiles with different silanes.
- Optimal lactadherin (LACT) concentration for uEV capture determined to be 25 µg/mL.
- Characterization of isolated uEVs' morphology and size distribution.
Conclusions:
- Optimized silicon surface functionalization protocols enhance uEV capture efficiency.
- Lactadherin at 25 µg/mL is effective for capturing uEVs on functionalized silicon surfaces.
- The developed methods provide a foundation for sensitive uEV-based diagnostics.
