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.
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
Keywords: Ratiometric sensing, Hyaluronidase, HA-FRET, Bladder cancer.
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