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Lifetime-based Sensing of the Hyaluronidase Using Fluorescein Labeled Hyaluronic Acid

Lifetime-based Sensing of the Hyaluronidase Using Fluorescein Labeled Hyaluronic Acid

Rafal Fudala, Mark E Mummert, Zygmunt Gryczynski, Ryan Rich, Julian Borejdo, Ignacy Gryczynski

J Photochem Photobiol B. 2012 Jan 5;106(1):69-73.

PMID: 22082776

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.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
LS741364	Fluorescein hyaluronic acid			Price

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