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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...

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Autores principales: Kamińska, Agnieszka, Marzec, Magdalena E., Stępień, Ewa Ł.
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
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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.
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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
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