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- Title
Label-free fluorescence turn-on detection of alkaline phosphatase activity using the calcein–Ce<sup>3+</sup> complex.
- Authors
Feng, Tingting; Huang, Yu; Yan, Shuzhu
- Abstract
This study introduces an innovative approach for the real-time and efficient detection of alkaline phosphatase (ALP) activity, using a calcein fluorescence probe and leveraging the static quenching properties of calcein fluorescence by Ce3+ metal ions. In this method, calcein serves as the signal element, with its fluorescence effectively preserved through energy transfer or charge transfer when coordinated with Ce3+. Conversely, ALP catalyzes the phosphopeptide substrate to generate a substantial amount of Pi, preventing calcein fluorescence quenching due to the higher affinity between Pi and Ce3+ compared with that between calcein and Ce3+. The fluorescence intensity ratio (F-F0/F0) exhibited excellent linearity, facilitating sensitive ALP detection. The proposed ALP detection method covers a range from 0 to 1.4 mU/mL (R2 = 0.9942), with the limit of detection at 0.069 mU/mL (S/N = 3). Additionally, this method was successfully applied for detecting ALP in serum samples and studying its inhibitors. This research introduces a novel clinical diagnosis approach for ALP sensing while broadening the potential applications of calcein.
- Publication
Analytical & Bioanalytical Chemistry, 2024, Vol 416, Issue 24, p5317
- ISSN
1618-2642
- Publication type
Article
- DOI
10.1007/s00216-024-05464-w