Ultrasensitive Fluorogenic Probe to Detect Heparanase Activity

Single-Step Fluorescence Assay Facilitates High-Throughput Screening of Heparanase Inhibitors

This fluorogenic probe tests for the presence of heparanase and facilitates rapid screening for heparanase inhibitors to aid the development of cancer therapeutics. Heparanase is an enzyme overexpressed in many carcinomas, sarcomas, hematological malignancies, and inflammatory conditions. Upregulation of heparanase expression correlates with increased tumor size, metastasis, and poor prognosis. Detecting heparanase in the body may inform cancer therapies. The cancer diagnostics market should reach $250 billion by 2026. Common heparanase assays either lack sensitivity and/or selectivity, or require multiple steps with multiple reagents, or work at unfavorable conditions.

 

Researchers at the University of Florida have developed an ultrasensitive and selective heparanase small-molecule fluorogenic probe. This one-step assay is sensitive only to heparanase, and its derivatives developed in the same lab allows for use with medical imaging systems such as MRI or PET scans.

 

 

Application

Ultrasensitive heparanase small molecule fluorogenic probe to detect heparanase and screen for its inhibitors

 

Advantages

  • Reliably detects heparanase, helping screen for it during the development of cancer therapeutics
  • High sensitivity, accurately detecting even low levels of heparanase
  • High selectivity, inhibiting interference from other similar biomolecules
  • Assay provides simple, rapid, one-step heparanase fluorescence detection

Technology

The fluorogenic heparanase probe is comprised of two easily divisible parts, a recognition unit, and a fluorophore molecule linked by a glycosidic bond to the reducing end of the recognition unit. When heparanase breaks the glycosidic bond, the probe releases the fluorophore, which produces fluorescence. This fluorescence can be seen with a fluorescence microscope or a regular plate reader. The recognition unit is a disaccharide based on heparanase’s bond cleavage patterns. An electron-withdrawing group increases fluorescence, allowing the assay to indicate heparanase activity.

Patent Information:
Title App Type Country Serial No. Patent No. File Date Issued Date Expire Date
Heparanase Compound and Methods of Use PCT from PRV PCT PCT/US2020/029627   4/23/2020   10/24/2021