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A Novel Technique for Generating and Observing Chemiluminescence in a Biological Setting

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Exploring a New Ruthenium(II) Complex with High DNA Binding Ability as a Novel Efficient Luminescent Intercalation

Yaoyao Xu¹, Rongxiu Deng¹, Junli Jia²

¹School of Chemistry and Life Sciences, Suzhou University of Science and Technology, Suzhou, Jiangsu 215009, China.

Analytical Chemistry

|February 25, 2025

Summary

This study developed a sensitive, label-free biosensor using a Ruthenium(II) complex and hybridization chain reaction (HCR) for enhanced DNA detection. The biosensor shows great potential for applications in anticancer and gene therapy.

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Area of Science:

Coordination Chemistry
Biotechnology
Biosensing

Background:

Ruthenium(II) complexes exhibit strong DNA binding, crucial for developing sensitive detection probes.
Hybridization chain reaction (HCR) amplifies DNA, creating platforms for enhanced signal generation.
Developing label-free biosensors is key for sensitive and efficient molecular detection.

Purpose of the Study:

To synthesize and characterize a Ruthenium(II) complex, Ru(dip)2(tpphz), for its DNA binding and luminescent properties.
To construct a label-free electrochemiluminescent/photoluminescent (ECL/PL) dual-mode biosensor utilizing HCR and the synthesized Ruthenium(II) complex.
To demonstrate the biosensor's capability for sensitive and selective target detection via signal modulation.

Main Methods:

Synthesis and characterization of Ru(dip)2(tpphz) complex.
Construction of a dual-mode biosensor involving magnetic silica spheres, trigger DNA, hairpin DNA, HCR, and Cas12a.
Utilizing UV-vis absorption spectroscopy and AutoDock for DNA binding analysis.
Employing ECL and PL measurements for signal detection.

Main Results:

Ru(dip)2(tpphz) confirmed high DNA binding capacity and dual ECL/PL properties.
The HCR-based biosensor achieved significant signal amplification.
The label-free biosensor demonstrated high selectivity and a low limit of detection (LOD) of 69 fM.
Cas12a activation led to trigger DNA cleavage, disappearance of HCR, and signal reduction.

Conclusions:

Ru(dip)2(tpphz) is a promising probe for label-free dual-mode biosensors due to its DNA binding and luminescent characteristics.
The HCR-amplified strategy significantly enhances detection sensitivity.
This approach holds substantial potential for applications in anticancer, gene therapy, and molecular probing.