Lifetime-based sensing of the hyaluronidase using fluorescein labeled hyaluronic acid

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10 Citations (Scopus)

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 languageEnglish
Pages (from-to)69-73
Number of pages5
JournalJournal of Photochemistry and Photobiology B: Biology
Volume106
Issue number1
DOIs
StatePublished - 5 Jan 2012

Fingerprint

Hyaluronoglucosaminidase
Hyaluronic Acid
Fluorescein
macromolecules
life (durability)
acids
Fluorescence
cleavage
quenching
fluorescence
Calibration
Amines
enzymes
amines
brightness
Enzymes
decay
approximation
molecules

Keywords

  • Fluorescence
  • Homo-FRET
  • Hyaluronan
  • Hyaluronidase
  • Lifetime based sensing

Cite this

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title = "Lifetime-based sensing of the hyaluronidase using fluorescein labeled hyaluronic acid",
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.",
keywords = "Fluorescence, Homo-FRET, Hyaluronan, Hyaluronidase, Lifetime based sensing",
author = "Rafal Fudala and Mummert, {Mark E.} and Zygmunt Gryczynski and Ryan Rich and Julian Borejdo and Ignacy Gryczynski",
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TY - JOUR

T1 - Lifetime-based sensing of the hyaluronidase using fluorescein labeled hyaluronic acid

AU - Fudala, Rafal

AU - Mummert, Mark E.

AU - Gryczynski, Zygmunt

AU - Rich, Ryan

AU - Borejdo, Julian

AU - Gryczynski, Ignacy

PY - 2012/1/5

Y1 - 2012/1/5

N2 - 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.

AB - 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.

KW - Fluorescence

KW - Homo-FRET

KW - Hyaluronan

KW - Hyaluronidase

KW - Lifetime based sensing

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U2 - 10.1016/j.jphotobiol.2011.10.005

DO - 10.1016/j.jphotobiol.2011.10.005

M3 - Article

C2 - 22082776

AN - SCOPUS:84155164483

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SP - 69

EP - 73

JO - Journal of Photochemistry and Photobiology, B: Biology

JF - Journal of Photochemistry and Photobiology, B: Biology

SN - 1011-1344

IS - 1

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