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Fluorescence detection of hyaluronidase

Paper ID Volume ID Publish Year Pages File Format Full-Text
30854 44508 2011 5 PDF Available
Title
Fluorescence detection of hyaluronidase
Abstract

We labeled hyaluronan (HA) with two fluorophores, fluorescein amine and rhodamine B amine. These two fluorophores are suitable for a fluorescence (Foerster) resonance energy transfer (FRET) which results in a fluorescein quenching and an enhanced rhodamine emission. Such labeled HA (HA–FRET) is a potential sensor for HA degradation. We studied fluorescence properties of HA–FRET in the absence and presence of hyaluronidase enzyme (HA-ase). The time-resolved fluorescence measurements indicate more than 50% of FRET in the absence of HA-ase. In the presence of HA-ase FRET decreases with time, and relative fluorescence intensities of fluorescein and rhodamine shifts to fluorescein indicating a release of FRET. The kinetics of the digestion process of HA by HA-ase depends on the concentration of the enzyme. We demonstrate that simultaneous measurements of green and red emission of HA–FRET can be used in ratio metric detection of the HA-ase presence and activity. This in turn, can be utilized for the construction of a robust but reliable HA-ase sensing device.

► We describe FRET-based detection of hyaluronidase. ► Spectroscopic evaluation of HA–FRET using steady state/time resolved fluorescence. ► Propose that HA–FRET is a useful substrate for HA-ase sensing.

Keywords
Hyaluronan; FRET; Hyaluronidase; Fluorescence; Ratio metric sensing
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Fluorescence detection of hyaluronidase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Photochemistry and Photobiology B: Biology - Volume 104, Issue 3, 2 September 2011, Pages 473–477
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us