FRET Based Ratio-Metric Sensing of Hyaluronidase in Synthetic Urine as a Biomarker for Bladder and Prostate Cancer
Rahul Chib, Sangram Raut, Rafal Fudala, Aaron Chang, Mark Mummert, Ryan Rich, Zygmunt Gryczynski and Ignacy Gryczynski
Affiliation: Department of Molecular Biology and Immunology, Center for Commercialization of Fluorescence Technologies, University of North Texas Health Science Center, Fort Worth, TX 76107, USA.
Keywords: Ratiometric sensing, Hyaluronidase, HA-FRET, Bladder cancer.
Elevated hyaluronidase levels are found in the urine of bladder and prostate cancer patients. Therefore, HA-ase
is regarded as an important biomarker for the detection of these cancers. In this report, we use a FRET based ratiometric
sensing approach to detect the level of HA-ase in synthetic urine. For this, we have used a HA-FRET probe (hyaluronan)
labeled with fluorescein as a donor and rhodamine as an acceptor. We monitor the digestion of our HA-FRET probe with
different concentrations of HA-ase in synthetic urine via fluorescence emission. The extent to which FRET is released depends
on the concentration of HA-ase. Our fluorescence intensity results are also supported with time resolved fluorescence
decay data. This assay can be used to develop a non-invasive technique for the detection of bladder and/or prostate
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