Journal Article Epublish 1 citation
PubMed: 40264865 DOI: 10.1039/d4ra08926j
Functionalized screen-printed electrodes for non-invasive detection of vascular-endothelial cadherin in extracellular vesicles.
RSC Adv 15(16):12609-12621
Authors: William Meza-Morales, Sahimy Ayus-Martinez, Jesus Jimenez-Osorio, Maria Buendia-Otero, Luis López, David Suleiman, Edu Suarez, Donald O Freytes, Lisandro Cunci, Camilo Mora
Affiliations
- Department of Chemical Engineering, University of Puerto Rico-Mayaguez Route 108 Mayaguez Puerto Rico USA.
- Department of Chemistry, University of Puerto Rico-Rio Piedras 601 Av. Universidad San Juan Puerto Rico USA.
- Department of Biology, University of Puerto Rico-Ponce Av. Santiago de los Caballeros Ponce Puerto Rico USA.
- Lampe Joint Department of Biomedical Engineering, The University of North Carolina at Chapel Hill/North Carolina State University 4130 Engineering Building III, Campus Box 7115 Raleigh NC 27695 USA.
Abstract
In this study, we developed a biosensor using a gold screen-printed electrode (Au-SPE) functionalized with mercaptoundecanoic acid (MUA) and an antibody for detecting the vascular-endothelial cadherin (CD144) as a endothelial biomarker protein on extracellular vesicles (EVs) isolated from saliva. The MUA functionalization provides a stable platform for immobilizing the CD144 antibody, ensuring the detection of the target protein. This biosensor combines Au-SPE technology with an immunoassay, offering a rapid, sensitive, and non-invasive method for detection of CD144 carried by EVs. Characterization of saliva-derived EVs using transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA) confirmed their morphology and size, which fell within the expected range of 80-180 nm. NTA indicated a lower concentration of particles in saliva-EVs than in serum-EVs (controls), highlighting the need for sensitive detection of EV cargos in this type of EV. Immunodetection confirmed the presence of CD144 in both saliva and serum-derived EVs, with higher concentrations in serum. Functionalization of Au-SPEs with MUA and CD144 antibodies was confirmed by significant resistance changes, and atomic force microscopy (AFM) was used to verify the preservation of EV morphology and their capturing post-immune adsorption. A calibration curve demonstrated the high sensitivity of the biosensor prototype for detecting CD144-positive EVs, with a limit of detection (LOD) of 0.111 ng mL and a limit of quantification (LOQ) of 0.37 ng mL, requiring only 3 μL of EV-sample. This biosensor shows potential as a novel method for detecting and studying endothelial biomarkers associated with cardiovascular disease in EVs isolated from saliva, a capability not currently available with existing tools. Furthermore, it provides a key platform for expanding research to other biomarkers and diseases by monitoring protein cargos in the EVs, enhancing its utility across diverse clinical applications.
MeSH
Extracellular Vesicles, Microscopy, Atomic Force, Serum, Electrodes, Cadherins, Biomarkers, Microscopy, Cadherin 5