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Direct Detection of Isolevuglandins in Tissues Using a D11 scFv-Alkaline Phosphatase Fusion Protein and Immunofluorescence
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A dual-mode biosensor for alkaline phosphatase detection based on DCIP and Eu3+-calcein
Bingyu Fu1,2,3, Yanru Shao1, Zhixiong Zheng1
1College of Life Science, Yangtze University, Jingzhou, Hubei, 434025, China. xuqi@yangtzeu.edu.cn.
Analytical Methods : Advancing Methods and Applications
|February 12, 2026
Summary
This study introduces a dual-mode biosensor for detecting alkaline phosphatase (ALP). The biosensor utilizes colorimetric and fluorescent signals, offering sensitive and selective detection of ALP in samples.
Area of Science:
- Biomedical Engineering
- Analytical Chemistry
- Biochemistry
Background:
- Alkaline phosphatase (ALP) is a crucial enzyme involved in various physiological and pathological processes.
- Accurate detection of ALP is essential for diagnosing diseases like liver disorders and bone diseases.
- Existing biosensors often lack sensitivity, selectivity, or dual-mode detection capabilities.
Purpose of the Study:
- To develop a simple and sensitive dual-mode biosensor for alkaline phosphatase (ALP) detection.
- To leverage both colorimetric and fluorescent signals for enhanced detection accuracy.
- To validate the biosensor's performance in complex biological matrices like serum samples.
Main Methods:
- Designed a dual-mode biosensor utilizing 2,6-dichloroindiphenol (DCIP) colorimetry and calcein fluorescence.
- Employed L-ascorbic acid-2-phosphate (AAP) as a substrate, which is dephosphorylated by ALP into ascorbic acid (AA) and phosphate ions.
- Utilized the reducing property of AA to decolorize DCIP and the chelating ability of phosphate ions to restore Eu3+-calcein fluorescence.
Main Results:
- Colorimetric mode: Linear detection range of 1.56-50 U L-1, Limit of Detection (LOD) of 0.27 U L-1.
- Fluorescence mode: Linear detection range of 0.78-25 U L-1, LOD of 0.23 U L-1.
- Demonstrated excellent selectivity for ALP and satisfactory recovery rates in serum samples.
Conclusions:
- The developed dual-mode biosensor offers a simple, sensitive, and selective method for ALP detection.
- The combined colorimetric and fluorescent approach enhances detection reliability.
- This biosensor shows potential for clinical diagnostics and biochemical analysis.
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