Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime Measurements
Interactions of nanoparticles with biomaterials determine the biological activity that is key for the physiological response. Dendritic polyglycerol sulfates (dPGS) were found recently to act as an inhibitor of inflammation by blocking selectins. Systemic application of dPGS would present this nanop...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2015-12-01
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| Series: | Molecules |
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| Online Access: | http://www.mdpi.com/1420-3049/21/1/22 |
| _version_ | 1818607807371411456 |
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| author | Alexander Boreham Jens Pikkemaat Pierre Volz Robert Brodwolf Christian Kuehne Kai Licha Rainer Haag Jens Dernedde Ulrike Alexiev |
| author_facet | Alexander Boreham Jens Pikkemaat Pierre Volz Robert Brodwolf Christian Kuehne Kai Licha Rainer Haag Jens Dernedde Ulrike Alexiev |
| author_sort | Alexander Boreham |
| collection | DOAJ |
| description | Interactions of nanoparticles with biomaterials determine the biological activity that is key for the physiological response. Dendritic polyglycerol sulfates (dPGS) were found recently to act as an inhibitor of inflammation by blocking selectins. Systemic application of dPGS would present this nanoparticle to various biological molecules that rapidly adsorb to the nanoparticle surface or lead to adsorption of the nanoparticle to cellular structures such as lipid membranes. In the past, fluorescence lifetime measurements of fluorescently tagged nanoparticles at a molecular and cellular/tissue level have been proven to reveal valuable information on the local nanoparticle environment via characteristic fluorescent lifetime signatures of the nanoparticle bound dye. Here, we established fluorescence lifetime measurements as a tool to determine the binding affinity to fluorescently tagged dPGS (dPGS-ICC; ICC: indocarbocyanine). The binding to a cell adhesion molecule (L-selectin) and a human complement protein (C1q) to dPGS-ICC was evaluated by the concentration dependent change in the unique fluorescence lifetime signature of dPGS-ICC. The apparent binding affinity was found to be in the nanomolar range for both proteins (L-selectin: 87 ± 4 nM and C1q: 42 ± 12 nM). Furthermore, the effect of human serum on the unique fluorescence lifetime signature of dPGS-ICC was measured and found to be different from the interactions with the two proteins and lipid membranes. A comparison between the unique lifetime signatures of dPGS-ICC in different biological environments shows that fluorescence lifetime measurements of unique dPGS-ICC fluorescence lifetime signatures are a versatile tool to probe the microenvironment of dPGS in cells and tissue. |
| first_indexed | 2024-12-16T14:32:37Z |
| format | Article |
| id | doaj.art-5d243541840f4c30b283c731f3e4768e |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-16T14:32:37Z |
| publishDate | 2015-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-5d243541840f4c30b283c731f3e4768e2022-12-21T22:28:12ZengMDPI AGMolecules1420-30492015-12-012112210.3390/molecules21010022molecules21010022Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime MeasurementsAlexander Boreham0Jens Pikkemaat1Pierre Volz2Robert Brodwolf3Christian Kuehne4Kai Licha5Rainer Haag6Jens Dernedde7Ulrike Alexiev8Institut für Experimentalphysik, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, GermanyInstitut für Experimentalphysik, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, GermanyInstitut für Experimentalphysik, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, GermanyInstitut für Experimentalphysik, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, GermanyInstitut für Laboratoriumsmedizin, Klinische Chemie und Pathobiochemie, Charité—Universitätsmedizin Berlin, Augustenburger Platz 1, 13353 Berlin, GermanyMivenion GmbH, Robert-Koch-Platz 4, 10115 Berlin, GermanyHelmholtz Virtual Institute—Multifunctional Biomaterials for Medicine, Helmholtz-Zentrum Geesthacht, Kantstr. 55, 14513 Teltow, GermanyHelmholtz Virtual Institute—Multifunctional Biomaterials for Medicine, Helmholtz-Zentrum Geesthacht, Kantstr. 55, 14513 Teltow, GermanyInstitut für Experimentalphysik, Freie Universität Berlin, Arnimallee 14, 14195 Berlin, GermanyInteractions of nanoparticles with biomaterials determine the biological activity that is key for the physiological response. Dendritic polyglycerol sulfates (dPGS) were found recently to act as an inhibitor of inflammation by blocking selectins. Systemic application of dPGS would present this nanoparticle to various biological molecules that rapidly adsorb to the nanoparticle surface or lead to adsorption of the nanoparticle to cellular structures such as lipid membranes. In the past, fluorescence lifetime measurements of fluorescently tagged nanoparticles at a molecular and cellular/tissue level have been proven to reveal valuable information on the local nanoparticle environment via characteristic fluorescent lifetime signatures of the nanoparticle bound dye. Here, we established fluorescence lifetime measurements as a tool to determine the binding affinity to fluorescently tagged dPGS (dPGS-ICC; ICC: indocarbocyanine). The binding to a cell adhesion molecule (L-selectin) and a human complement protein (C1q) to dPGS-ICC was evaluated by the concentration dependent change in the unique fluorescence lifetime signature of dPGS-ICC. The apparent binding affinity was found to be in the nanomolar range for both proteins (L-selectin: 87 ± 4 nM and C1q: 42 ± 12 nM). Furthermore, the effect of human serum on the unique fluorescence lifetime signature of dPGS-ICC was measured and found to be different from the interactions with the two proteins and lipid membranes. A comparison between the unique lifetime signatures of dPGS-ICC in different biological environments shows that fluorescence lifetime measurements of unique dPGS-ICC fluorescence lifetime signatures are a versatile tool to probe the microenvironment of dPGS in cells and tissue.http://www.mdpi.com/1420-3049/21/1/22nanomedicinedendritic polymersprotein coronafluorescence lifetime |
| spellingShingle | Alexander Boreham Jens Pikkemaat Pierre Volz Robert Brodwolf Christian Kuehne Kai Licha Rainer Haag Jens Dernedde Ulrike Alexiev Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime Measurements Molecules nanomedicine dendritic polymers protein corona fluorescence lifetime |
| title | Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime Measurements |
| title_full | Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime Measurements |
| title_fullStr | Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime Measurements |
| title_full_unstemmed | Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime Measurements |
| title_short | Detecting and Quantifying Biomolecular Interactions of a Dendritic Polyglycerol Sulfate Nanoparticle Using Fluorescence Lifetime Measurements |
| title_sort | detecting and quantifying biomolecular interactions of a dendritic polyglycerol sulfate nanoparticle using fluorescence lifetime measurements |
| topic | nanomedicine dendritic polymers protein corona fluorescence lifetime |
| url | http://www.mdpi.com/1420-3049/21/1/22 |
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