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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| Language: | English |
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MDPI AG
2021-08-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/26/16/4764 |
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| author | Agnieszka Kamińska Magdalena E. Marzec Ewa Ł. Stępień |
| author_facet | Agnieszka Kamińska Magdalena E. Marzec Ewa Ł. Stępień |
| author_sort | Agnieszka Kamińska |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-03-10T08:32:52Z |
| format | Article |
| id | doaj.art-173bdbb6a42742b19e72c03e4af087df |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T08:32:52Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-173bdbb6a42742b19e72c03e4af087df2023-11-22T08:51:37ZengMDPI AGMolecules1420-30492021-08-012616476410.3390/molecules26164764Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular VesiclesAgnieszka Kamińska0Magdalena E. Marzec1Ewa Ł. Stępień2Department of Medical Physics, Marian Smoluchowski Institute of Physics, Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, 30-348 Kraków, PolandDepartment of Medical Physics, Marian Smoluchowski Institute of Physics, Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, 30-348 Kraków, PolandDepartment of Medical Physics, Marian Smoluchowski Institute of Physics, Faculty of Physics, Astronomy and Applied Computer Science, Jagiellonian University, 30-348 Kraków, PolandFor 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.https://www.mdpi.com/1420-3049/26/16/4764extracellular vesicleslactadherinToF-SIMS |
| spellingShingle | Agnieszka Kamińska Magdalena E. Marzec Ewa Ł. Stępień Design and Optimization of a Biosensor Surface Functionalization to Effectively Capture Urinary Extracellular Vesicles Molecules extracellular vesicles lactadherin ToF-SIMS |
| 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 | extracellular vesicles lactadherin ToF-SIMS |
| url | https://www.mdpi.com/1420-3049/26/16/4764 |
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