A fluorometric assay for alkaline phosphatase activity based on β-cyclodextrin-modified carbon quantum dots through...

Area of Science:

• Biochemistry
• Nanotechnology
• Analytical Chemistry

Background:

• Alkaline phosphatase (ALP) is a crucial enzyme in clinical diagnostics.
• Developing sensitive and reliable assays for ALP activity is essential for disease monitoring.
• Existing methods may lack real-time capabilities or sensitivity.

Purpose of the Study:

• To develop a convenient, reliable, and highly sensitive real-time assay for alkaline phosphatase (ALP) activity.
• To utilize β-cyclodextrin-modified carbon quantum dots (β-CD-CQDs) as a nanoprobe for enzyme detection.
• To establish a quantitative evaluation method for ALP activity based on host-guest recognition and fluorescence quenching.

Main Methods:

• Synthesized β-cyclodextrin-modified carbon quantum dots (β-CD-CQDs) nanoprobe.
• Employed p-nitrophenol phosphate disodium salt as the substrate for ALP.
• Utilized host-guest recognition and photoinduced electron transfer (PET) for fluorescence quenching.
• Established quantitative evaluation of ALP activity through fluorescence intensity correlation.

Main Results:

• Developed a β-CD-CQDs nanoprobe for ALP detection.
• Achieved real-time monitoring of ALP activity with high sensitivity.
• Quantified ALP activity in a broad range (3.4–100.0 U/L) with a detection limit of 0.9 U/L.
• Demonstrated high specificity for ALP, even in the presence of high glucose concentrations.

Conclusions:

• The developed β-CD-CQDs nanoprobe provides a sensitive and reliable method for real-time ALP activity detection.
• The assay leverages host-guest recognition and PET for efficient fluorescence quenching.
• This approach broadens the application of host-guest recognition for enzyme detection in clinical practice.