Abstract
In this report we propose a lifetime-based sensing (LBS) for the detection of hyaluronidase (HA-ase). First, we heavily label hyaluronan macromolecules (HAs) with fluorescein amine. The fluorescein labeled HA (HA-Fl) has a weak fluorescence and short fluorescence lifetime due to an efficient self-quenching. Upon the addition of HA-ase, the brightness and lifetime of the sample increase. The cleavage of an HA macromolecule reduces the energy migration between fluorescein molecules and the degree of the self-quenching. A first order of the cleavage reaction depends on the amount of the HA-ase enzyme. We describe an HA-ase sensing strategy based on the lifetime changes of the fluorescein labeled HA in the presence of HA-ase. We demonstrate that the calibration of the sensing response is the same for the average lifetime as for a single exponential decay approximation, which significantly simplifies the analysis of the sensing measurements.
Original language | English |
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Pages (from-to) | 69-73 |
Number of pages | 5 |
Journal | Journal of Photochemistry and Photobiology B: Biology |
Volume | 106 |
Issue number | 1 |
DOIs | |
State | Published - 5 Jan 2012 |
Keywords
- Fluorescence
- Homo-FRET
- Hyaluronan
- Hyaluronidase
- Lifetime based sensing