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Colorimetric Analysis of Alkaline Phosphatase Activity in S. aureus Biofilm
Published on: April 12, 2019
A gold nanoparticles-based colorimetric assay for alkaline phosphatase detection with tunable dynamic range
Chun Mei Li1, Shu Jun Zhen, Jian Wang
1Education Ministry Key Laboratory on Luminescence and Real-Time Analysis, College of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
Biosensors & Bioelectronics
|January 30, 2013
Summary
A new colorimetric assay detects alkaline phosphatase (ALP) using gold nanoparticles (AuNPs) and adenosine triphosphate (ATP). This label-free method offers sensitive detection with a tunable range, even in human serum.
Area of Science:
- * Nanotechnology and Nanoscience
- * 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 methods for ALP detection can be complex, time-consuming, or lack sensitivity.
Purpose of the Study:
- * To develop a simple, label-free, and highly sensitive colorimetric assay for ALP detection.
- * To utilize a gold nanoparticle/adenosine triphosphate (AuNP/ATP) sensing system for visual and instrumental ALP quantification.
- * To demonstrate the adaptability of the assay's dynamic range and its applicability in complex biological samples.
Main Methods:
- * Fabrication of cetyltrimethylammonium bromide (CTAB)-capped gold nanoparticles (AuNPs).
- * Design of an assay based on ATP-induced AuNP aggregation and ALP-mediated disaggregation.
- * Colorimetric and surface plasmon resonance (SPR) measurements for monitoring ALP activity.
- * Adjustment of the assay's dynamic range using different metal ions (Ca(2+), Pb(2+)).
Main Results:
- * The assay exhibited high sensitivity and selectivity for ALP detection.
- * A distinct visual color change (gray-to-red) was observed in the presence of ALP.
- * The SPR absorption band shift correlated with ALP concentration, enabling quantitative analysis.
- * The dynamic range of the assay was successfully tuned (e.g., 5.0–100 U/L with Ca(2+), 0.2–20 U/L with Pb(2+)).
- * The assay demonstrated consistent performance in human serum samples.
Conclusions:
- * A novel, label-free colorimetric assay for ALP detection based on AuNP/ATP interactions was successfully developed.
- * The assay is sensitive, selective, visually observable, and offers a tunable dynamic range.
- * The method's robustness in human serum samples highlights its potential for clinical diagnostics.
