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Competitive host-guest interaction between β-cyclodextrin polymer and pyrene-labeled probes for fluorescence analyses
Pei Liu1, Shan Sun, Xiaochen Guo
1State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University , Changsha, Hunan 410082, China.
Analytical Chemistry
|January 28, 2015
Summary
We developed a novel fluorescence assay using a competitive host-guest interaction mechanism for sensitive DNA detection. This method offers a clear signal turn-off for target DNA identification, with potential for broader biochemical applications.
Area of Science:
- Biochemistry
- Analytical Chemistry
- Molecular Biology
Background:
- Homogeneous fluorescence analysis offers advantages in biochemical assays.
- Host-guest interactions provide a basis for molecular recognition and signal transduction.
- Cyclodextrin polymers are effective hosts for various guest molecules.
Purpose of the Study:
- To develop a novel homogeneous fluorescence analysis based on competitive host-guest interaction (CHGI).
- To create a sensitive detection system for target DNA using pyrene-labeled probes and β-cyclodextrin polymer (polyβ CD).
- To explore the potential for detecting other biomolecules, such as adenosine.
Main Methods:
- Utilized a competitive host-guest interaction (CHGI) mechanism between polyβ CD and pyrene-labeled oligonucleotide probes.
- Employed pyrene fluorescence as a reporter, with luminescence intensity modulated by polyβ CD binding.
- Incorporated Exodeoxyribonuclease I (Exo I) and Exodeoxyribonuclease III (Exo III) to enhance assay sensitivity.
Main Results:
- Achieved enhanced pyrene luminescence due to favorable microenvironment within polyβ CD cavities.
- Demonstrated a clear fluorescence turn-off signal upon target DNA binding, hindering pyrene entry into polyβ CD.
- Showcased enhanced sensitivity through enzymatic cleavage and subsequent stronger host-guest interaction of pyrene-nucleotides.
- Successfully detected adenosine using a similar sensing strategy.
Conclusions:
- The developed CHGI-based fluorescence assay provides a sensitive and specific method for DNA detection.
- The system exhibits potential for detecting a wide range of aptamer-binding chemicals and biomolecules.
- Further optimization may be needed to mitigate interference from complex biological samples.
