Fluorescent biosensor for the detection of hyaluronidase

intensity-based ratiometric sensing and fluorescence lifetime-based sensing using a long lifetime azadioxatriangulenium (ADOTA) fluorophore

Rahul Chib, Mark Mummert, Ilkay Bora, Bo W. Laursen, Sunil Shah, Robert Pendry, Ignacy Gryczynski, Julian Borejdo, Zygmunt Gryczynski, Rafal Fudala

Research output: Contribution to journalArticleResearchpeer-review

6 Citations (Scopus)

Abstract

In this report, we have designed a rapid and sensitive, intensity-based ratiometric sensing as well as lifetime-based sensing probe for the detection of hyaluronidase activity. Hyaluronidase expression is known to be upregulated in various pathological conditions. We have developed a fluorescent probe by heavy labeling of hyaluronic acid with a new orange/red-emitting organic azadioxatriangulenium (ADOTA) fluorophore, which exhibits a long fluorescence lifetime (∼20 ns). The ADOTA fluorophore in water has a peak fluorescence lifetime of ∼20 ns and emission spectra centered at 560 nm. The heavily ADOTA-labeled hyaluronic acid (HA-ADOTA) shows a red shift in the peak emission wavelength (605 nm), a weak fluorescence signal, and a shorter fluorescence lifetime (∼4 ns) due to efficient self-quenching and formation of aggregates. In the presence of hyaluronidase, the brightness and fluorescence lifetime of the sample increase with a blue shift in the peak emission to its original wavelength at 560 nm. The ratio of the fluorescence intensity of the HA-ADOTA probe at 560 and 605 nm can be used as the sensing method for the detection of hyaluronidase. The cleavage of the hyaluronic acid macromolecule reduces the energy migration between ADOTA molecules, as well as the degree of self-quenching and aggregation. This probe can be efficiently used for both intensity-based ratiometric sensing as well as fluorescence lifetime-based sensing of hyaluronidase. The proposed method makes it a rapid and sensitive assay, useful for analyzing levels of hyaluronidase in relevant clinical samples like urine or plasma. [Figure not available: see fulltext.]

Original languageEnglish
Pages (from-to)3811-3821
Number of pages11
JournalAnalytical and Bioanalytical Chemistry
Volume408
Issue number14
DOIs
StatePublished - 1 May 2016

Fingerprint

Hyaluronoglucosaminidase
Fluorophores
Biosensing Techniques
Biosensors
Fluorescence
Hyaluronic Acid
Quenching
Wavelength
Macromolecules
Fluorescent Dyes
Labeling
Luminance
Assays
Agglomeration
Urine
Plasmas
Molecules
Water

Keywords

  • Azadioxatriangulenium fluorophore
  • Fluorescence lifetime-based sensing
  • Fluorescence-based assay
  • Hyaluronidase sensing
  • Ratiometric sensing

Cite this

@article{b8bb7405e9da43e7b3624fd2400006ad,
title = "Fluorescent biosensor for the detection of hyaluronidase: intensity-based ratiometric sensing and fluorescence lifetime-based sensing using a long lifetime azadioxatriangulenium (ADOTA) fluorophore",
abstract = "In this report, we have designed a rapid and sensitive, intensity-based ratiometric sensing as well as lifetime-based sensing probe for the detection of hyaluronidase activity. Hyaluronidase expression is known to be upregulated in various pathological conditions. We have developed a fluorescent probe by heavy labeling of hyaluronic acid with a new orange/red-emitting organic azadioxatriangulenium (ADOTA) fluorophore, which exhibits a long fluorescence lifetime (∼20 ns). The ADOTA fluorophore in water has a peak fluorescence lifetime of ∼20 ns and emission spectra centered at 560 nm. The heavily ADOTA-labeled hyaluronic acid (HA-ADOTA) shows a red shift in the peak emission wavelength (605 nm), a weak fluorescence signal, and a shorter fluorescence lifetime (∼4 ns) due to efficient self-quenching and formation of aggregates. In the presence of hyaluronidase, the brightness and fluorescence lifetime of the sample increase with a blue shift in the peak emission to its original wavelength at 560 nm. The ratio of the fluorescence intensity of the HA-ADOTA probe at 560 and 605 nm can be used as the sensing method for the detection of hyaluronidase. The cleavage of the hyaluronic acid macromolecule reduces the energy migration between ADOTA molecules, as well as the degree of self-quenching and aggregation. This probe can be efficiently used for both intensity-based ratiometric sensing as well as fluorescence lifetime-based sensing of hyaluronidase. The proposed method makes it a rapid and sensitive assay, useful for analyzing levels of hyaluronidase in relevant clinical samples like urine or plasma. [Figure not available: see fulltext.]",
keywords = "Azadioxatriangulenium fluorophore, Fluorescence lifetime-based sensing, Fluorescence-based assay, Hyaluronidase sensing, Ratiometric sensing",
author = "Rahul Chib and Mark Mummert and Ilkay Bora and Laursen, {Bo W.} and Sunil Shah and Robert Pendry and Ignacy Gryczynski and Julian Borejdo and Zygmunt Gryczynski and Rafal Fudala",
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Fluorescent biosensor for the detection of hyaluronidase : intensity-based ratiometric sensing and fluorescence lifetime-based sensing using a long lifetime azadioxatriangulenium (ADOTA) fluorophore. / Chib, Rahul; Mummert, Mark; Bora, Ilkay; Laursen, Bo W.; Shah, Sunil; Pendry, Robert; Gryczynski, Ignacy; Borejdo, Julian; Gryczynski, Zygmunt; Fudala, Rafal.

In: Analytical and Bioanalytical Chemistry, Vol. 408, No. 14, 01.05.2016, p. 3811-3821.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - Fluorescent biosensor for the detection of hyaluronidase

T2 - intensity-based ratiometric sensing and fluorescence lifetime-based sensing using a long lifetime azadioxatriangulenium (ADOTA) fluorophore

AU - Chib, Rahul

AU - Mummert, Mark

AU - Bora, Ilkay

AU - Laursen, Bo W.

AU - Shah, Sunil

AU - Pendry, Robert

AU - Gryczynski, Ignacy

AU - Borejdo, Julian

AU - Gryczynski, Zygmunt

AU - Fudala, Rafal

PY - 2016/5/1

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N2 - In this report, we have designed a rapid and sensitive, intensity-based ratiometric sensing as well as lifetime-based sensing probe for the detection of hyaluronidase activity. Hyaluronidase expression is known to be upregulated in various pathological conditions. We have developed a fluorescent probe by heavy labeling of hyaluronic acid with a new orange/red-emitting organic azadioxatriangulenium (ADOTA) fluorophore, which exhibits a long fluorescence lifetime (∼20 ns). The ADOTA fluorophore in water has a peak fluorescence lifetime of ∼20 ns and emission spectra centered at 560 nm. The heavily ADOTA-labeled hyaluronic acid (HA-ADOTA) shows a red shift in the peak emission wavelength (605 nm), a weak fluorescence signal, and a shorter fluorescence lifetime (∼4 ns) due to efficient self-quenching and formation of aggregates. In the presence of hyaluronidase, the brightness and fluorescence lifetime of the sample increase with a blue shift in the peak emission to its original wavelength at 560 nm. The ratio of the fluorescence intensity of the HA-ADOTA probe at 560 and 605 nm can be used as the sensing method for the detection of hyaluronidase. The cleavage of the hyaluronic acid macromolecule reduces the energy migration between ADOTA molecules, as well as the degree of self-quenching and aggregation. This probe can be efficiently used for both intensity-based ratiometric sensing as well as fluorescence lifetime-based sensing of hyaluronidase. The proposed method makes it a rapid and sensitive assay, useful for analyzing levels of hyaluronidase in relevant clinical samples like urine or plasma. [Figure not available: see fulltext.]

AB - In this report, we have designed a rapid and sensitive, intensity-based ratiometric sensing as well as lifetime-based sensing probe for the detection of hyaluronidase activity. Hyaluronidase expression is known to be upregulated in various pathological conditions. We have developed a fluorescent probe by heavy labeling of hyaluronic acid with a new orange/red-emitting organic azadioxatriangulenium (ADOTA) fluorophore, which exhibits a long fluorescence lifetime (∼20 ns). The ADOTA fluorophore in water has a peak fluorescence lifetime of ∼20 ns and emission spectra centered at 560 nm. The heavily ADOTA-labeled hyaluronic acid (HA-ADOTA) shows a red shift in the peak emission wavelength (605 nm), a weak fluorescence signal, and a shorter fluorescence lifetime (∼4 ns) due to efficient self-quenching and formation of aggregates. In the presence of hyaluronidase, the brightness and fluorescence lifetime of the sample increase with a blue shift in the peak emission to its original wavelength at 560 nm. The ratio of the fluorescence intensity of the HA-ADOTA probe at 560 and 605 nm can be used as the sensing method for the detection of hyaluronidase. The cleavage of the hyaluronic acid macromolecule reduces the energy migration between ADOTA molecules, as well as the degree of self-quenching and aggregation. This probe can be efficiently used for both intensity-based ratiometric sensing as well as fluorescence lifetime-based sensing of hyaluronidase. The proposed method makes it a rapid and sensitive assay, useful for analyzing levels of hyaluronidase in relevant clinical samples like urine or plasma. [Figure not available: see fulltext.]

KW - Azadioxatriangulenium fluorophore

KW - Fluorescence lifetime-based sensing

KW - Fluorescence-based assay

KW - Hyaluronidase sensing

KW - Ratiometric sensing

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U2 - 10.1007/s00216-016-9472-5

DO - 10.1007/s00216-016-9472-5

M3 - Article

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SP - 3811

EP - 3821

JO - Analytical and Bioanalytical Chemistry

JF - Analytical and Bioanalytical Chemistry

SN - 1618-2642

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