Cargando…
Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles
For this study, we tested and optimized silicon surface functionalization procedures for capturing urinary extracellular vesicles (uEVs). The influence of the silane type (APTES or GOPS) and protein concentration on the efficiency of uEVs binding was investigated. Human lactadherin protein (LACT) wa...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399133/ https://www.ncbi.nlm.nih.gov/pubmed/34443351 http://dx.doi.org/10.3390/molecules26164764 |
_version_ | 1783745002788093952 |
---|---|
author | Kamińska, Agnieszka Marzec, Magdalena E. Stępień, Ewa Ł. |
author_facet | Kamińska, Agnieszka Marzec, Magdalena E. Stępień, Ewa Ł. |
author_sort | Kamińska, Agnieszka |
collection | PubMed |
description | For this study, we tested and optimized silicon surface functionalization procedures for capturing urinary extracellular vesicles (uEVs). The influence of the silane type (APTES or GOPS) and protein concentration on the efficiency of uEVs binding was investigated. Human lactadherin protein (LACT) was used to capture uEVs. We applied surface characterization techniques, including ellipsometry, atomic force microscopy, and time-of-flight secondary ion mass spectrometry, to observe changes in the biosensor surface after each functionalization step. uEVs were purified by a low-vacuum filtration method and concentrated by ultracentrifugation. The physical parameters of uEVs after the isolation procedure, such as morphology and size distribution, were determined using transmission electron microscopy and tunable resistive pulse sensing methods. We observed a gradual growth of the molecular layer after subsequent stages of modification of the silicon surface. The ToF-SIMS results showed no changes in the mean intensities for the characteristic peaks of amino acids and lipids in positive and negative polarization, in terms of the surface-modifying silane (APTES or GOPS) used. The most optimal concentration of LACT for the tested system was 25 µg/mL. |
format | Online Article Text |
id | pubmed-8399133 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-83991332021-08-29 Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles Kamińska, Agnieszka Marzec, Magdalena E. Stępień, Ewa Ł. Molecules Article For this study, we tested and optimized silicon surface functionalization procedures for capturing urinary extracellular vesicles (uEVs). The influence of the silane type (APTES or GOPS) and protein concentration on the efficiency of uEVs binding was investigated. Human lactadherin protein (LACT) was used to capture uEVs. We applied surface characterization techniques, including ellipsometry, atomic force microscopy, and time-of-flight secondary ion mass spectrometry, to observe changes in the biosensor surface after each functionalization step. uEVs were purified by a low-vacuum filtration method and concentrated by ultracentrifugation. The physical parameters of uEVs after the isolation procedure, such as morphology and size distribution, were determined using transmission electron microscopy and tunable resistive pulse sensing methods. We observed a gradual growth of the molecular layer after subsequent stages of modification of the silicon surface. The ToF-SIMS results showed no changes in the mean intensities for the characteristic peaks of amino acids and lipids in positive and negative polarization, in terms of the surface-modifying silane (APTES or GOPS) used. The most optimal concentration of LACT for the tested system was 25 µg/mL. MDPI 2021-08-06 /pmc/articles/PMC8399133/ /pubmed/34443351 http://dx.doi.org/10.3390/molecules26164764 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Kamińska, Agnieszka Marzec, Magdalena E. Stępień, Ewa Ł. Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles |
title | Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles |
title_full | Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles |
title_fullStr | Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles |
title_full_unstemmed | Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles |
title_short | Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles |
title_sort | design and optimization of a biosensor surface functionalization to effectively capture urinary extracellular vesicles |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8399133/ https://www.ncbi.nlm.nih.gov/pubmed/34443351 http://dx.doi.org/10.3390/molecules26164764 |
work_keys_str_mv | AT kaminskaagnieszka designandoptimizationofabiosensorsurfacefunctionalizationtoeffectivelycaptureurinaryextracellularvesicles AT marzecmagdalenae designandoptimizationofabiosensorsurfacefunctionalizationtoeffectivelycaptureurinaryextracellularvesicles AT stepienewał designandoptimizationofabiosensorsurfacefunctionalizationtoeffectivelycaptureurinaryextracellularvesicles |