A fluorescent probe for monitoring beta-secretase activity in cells

Unmet Need

Alzheimer’s Disease (AD) is characterized by the accumulation of amyloid-beta (Aβ) plaques, which lead to neurotoxicity, synaptic damage, and neuronal cell death. Current therapeutic approaches face significant challenges due to the complex nature of amyloid precursor protein (APP) processing, particularly the role of β-secretase (BACE). Effective BACE inhibitors have proven difficult to develop, primarily due to issues with bioavailability, metabolic clearance, and the enzyme's cellular localization. Existing assays for evaluating BACE inhibitors in cellular models are labor-intensive, slow, and often require genetic modifications, leaving a gap for more efficient methods to monitor BACE activity in live cells and organisms.

Technology Description

Duke inventors have developed a novel molecular probe designed to image β-secretase (BACE) activity in real time within living cells and organisms. The probe consists of a fluorescent group, a BACE substrate module, a quenching group, linker molecules, and a cell membrane anchor. This probe utilizes a Fluorescence Resonance Energy Transfer (FRET) mechanism, allowing for detection of BACE activity through changes in fluorescence upon substrate cleavage. By enabling the imaging of BACE activity without the need for genetically modified cell lines or antibodies, this technology offers a significant advancement in AD research.

Advantages

• Provides the capability to visualize BACE activity in live cells and organisms
• Eliminates the need for time-consuming and costly genetic manipulations
• The FRET-based design enhances sensitivity in detecting BACE activity, facilitating earlier and more accurate assessments of inhibitor efficacy
• Can be used in various experimental setups
• Reduces the complexity and labor intensity of traditional ELISA assays, enabling high-throughput screening of BACE inhibitors