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Label-Free Fluorescence-Based Enzyme Assays Using Supramolecular Host-Guest Approaches
Zhe Zheng1, Yiran Xu2, Ruina Liu1
1Tianjin Key Laboratory of Structure and Performance for Functional Molecules, College of Chemistry, Tianjin Normal University, Tianjin, P. R. China.
Small Methods
|February 18, 2026
Summary
Supramolecular tandem enzyme assays (STA) offer a simple, sensitive, and specific method for real-time monitoring of enzymatic reactions. This review highlights STA advancements for enzyme kinetics, inhibitor screening, and biomarker analysis.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Supramolecular Chemistry
Background:
- Enzymatic reaction monitoring is vital across enzymology, drug metabolism, and bioengineering.
- Supramolecular tandem enzyme assays (STA) utilize indicator displacement assays and differential host-guest binding.
- STA offers label-free, real-time, rapid, sensitive, and specific monitoring of enzyme activity.
Purpose of the Study:
- To review recent advancements in supramolecular tandem enzyme assay (STA) strategies.
- To cover diverse enzyme types and applications of STA.
- To identify current challenges and future perspectives in STA development.
Main Methods:
- Leveraging differential binding affinities of supramolecular hosts for substrates and products.
- Utilizing indicator displacement assays for signal generation.
- Applying STA for label-free, real-time reaction monitoring.
Main Results:
- STA has been widely applied for determining enzymatic kinetic parameters.
- STA is effective for screening enzyme inhibitors and analyzing enantiomeric excess.
- STA serves as a valuable tool for diagnosing biomarkers.
Conclusions:
- STA represents a powerful and versatile platform for enzymatic reaction analysis.
- Continued development of STA strategies will enhance enzyme studies and diagnostics.
- Future research should address current challenges to further expand STA applications.
