Immobilized Fluorescent Probes for Simultaneous Multiple Protease Detection
A new concept for simultaneous detection of two model proteases based on immobilized fluorescently labelled peptides was developed and evaluated. Each probe was composed of a carrier modified by poly(ethylene glycol) (PEG) chains, a specifically cleavable linker, and a fluorescent dye incorporated i...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2021-05-01
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| Series: | Chemosensors |
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| Online Access: | https://www.mdpi.com/2227-9040/9/6/119 |
| _version_ | 1797532967015284736 |
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| author | David Milićević Jan Hlaváč |
| author_facet | David Milićević Jan Hlaváč |
| author_sort | David Milićević |
| collection | DOAJ |
| description | A new concept for simultaneous detection of two model proteases based on immobilized fluorescently labelled peptides was developed and evaluated. Each probe was composed of a carrier modified by poly(ethylene glycol) (PEG) chains, a specifically cleavable linker, and a fluorescent dye incorporated in the peptide tail. Based on a single excitation–double emission fluorescence response of liberated fluorophores caused by enzymatic cleavage, the presence of a single or both proteases in a mixture was unambiguously determined in an experimentally established concentration range. Among the tested solid supports, Rink Amide PEGA resin was recognized as the most suitable choice from the perspective of on-resin enzyme assays. |
| first_indexed | 2024-03-10T11:07:54Z |
| format | Article |
| id | doaj.art-f59f7812041948409d833efffadd8063 |
| institution | Directory Open Access Journal |
| issn | 2227-9040 |
| language | English |
| last_indexed | 2024-03-10T11:07:54Z |
| publishDate | 2021-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Chemosensors |
| spelling | doaj.art-f59f7812041948409d833efffadd80632023-11-21T21:04:18ZengMDPI AGChemosensors2227-90402021-05-019611910.3390/chemosensors9060119Immobilized Fluorescent Probes for Simultaneous Multiple Protease DetectionDavid Milićević0Jan Hlaváč1Department of Organic Chemistry, Faculty of Science, Palacký University Olomouc, 17. Listopadu 12, 771 46 Olomouc, Czech RepublicDepartment of Organic Chemistry, Faculty of Science, Palacký University Olomouc, 17. Listopadu 12, 771 46 Olomouc, Czech RepublicA new concept for simultaneous detection of two model proteases based on immobilized fluorescently labelled peptides was developed and evaluated. Each probe was composed of a carrier modified by poly(ethylene glycol) (PEG) chains, a specifically cleavable linker, and a fluorescent dye incorporated in the peptide tail. Based on a single excitation–double emission fluorescence response of liberated fluorophores caused by enzymatic cleavage, the presence of a single or both proteases in a mixture was unambiguously determined in an experimentally established concentration range. Among the tested solid supports, Rink Amide PEGA resin was recognized as the most suitable choice from the perspective of on-resin enzyme assays.https://www.mdpi.com/2227-9040/9/6/119solid-phase synthesisimmobilized peptidefluorophorefluorescencesimultaneous detectionenzymatic cleavage |
| spellingShingle | David Milićević Jan Hlaváč Immobilized Fluorescent Probes for Simultaneous Multiple Protease Detection Chemosensors solid-phase synthesis immobilized peptide fluorophore fluorescence simultaneous detection enzymatic cleavage |
| title | Immobilized Fluorescent Probes for Simultaneous Multiple Protease Detection |
| title_full | Immobilized Fluorescent Probes for Simultaneous Multiple Protease Detection |
| title_fullStr | Immobilized Fluorescent Probes for Simultaneous Multiple Protease Detection |
| title_full_unstemmed | Immobilized Fluorescent Probes for Simultaneous Multiple Protease Detection |
| title_short | Immobilized Fluorescent Probes for Simultaneous Multiple Protease Detection |
| title_sort | immobilized fluorescent probes for simultaneous multiple protease detection |
| topic | solid-phase synthesis immobilized peptide fluorophore fluorescence simultaneous detection enzymatic cleavage |
| url | https://www.mdpi.com/2227-9040/9/6/119 |
| work_keys_str_mv | AT davidmilicevic immobilizedfluorescentprobesforsimultaneousmultipleproteasedetection AT janhlavac immobilizedfluorescentprobesforsimultaneousmultipleproteasedetection |