An activity-based fluorescent probe and its application for differentiating alkaline phosphatase activity in...

Area of Science:

• Biochemistry
• Chemical Biology
• Molecular Probes

Background:

• Alkaline phosphatase (ALP) is a crucial enzyme involved in various physiological and pathological processes.
• Developing selective and sensitive probes for ALP detection is essential for biological research and diagnostics.
• Existing probes may suffer from limitations such as low sensitivity, poor selectivity, or complex operation.

Purpose of the Study:

• To develop and characterize a novel fluorescent probe, AE-Phos, for the detection of ALP activity.
• To investigate the combined advantages of aggregation-induced emission (AIE) and excited state intramolecular proton transfer (ESIPT) in the probe design.
• To evaluate the probe's performance in quantitative imaging of ALP activity in cellular environments.

Main Methods:

• Synthesis and characterization of the AE-Phos fluorescent probe.
• Fluorescence spectroscopy to assess probe selectivity, sensitivity, and response kinetics towards ALP.
• Cellular experiments for quantitative imaging of ALP activity in different cell lines using the AE-Phos probe.

Main Results:

• AE-Phos demonstrated excellent selectivity and high sensitivity for ALP detection.
• The probe exhibited a large Stokes shift and a rapid response to ALP activity.
• Quantitative imaging of ALP activity in various cell lines was successfully achieved using AE-Phos.

Conclusions:

• AE-Phos is a highly effective fluorescent probe for detecting ALP activity, leveraging AIE and ESIPT characteristics.
• The probe offers significant advantages in terms of sensitivity, selectivity, and response time.
• AE-Phos provides a valuable tool for quantitative ALP activity imaging in biological systems.