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Aptasensor based on triplex switch for SERS detection of cytochrome c.

Yunfeng Xia¹, Peiyi Gao, Xiaowen Qiu

¹Life and Environmental Science College, Shanghai Normal University, Shanghai, 200234, P R China.

|October 27, 2012

Summary

We developed a sensitive surface-enhanced Raman scattering (SERS) method for detecting cytochrome c. This approach utilizes conformational changes in a signal transduction probe (STP) upon target binding, enhancing SERS signals for accurate detection.

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Area of Science:

Biomedical Engineering
Analytical Chemistry
Biosensing

Background:

Cytochrome c is a crucial biomarker in various physiological and pathological processes.
Sensitive and selective detection methods are needed for accurate cytochrome c quantification.
Existing biosensing techniques face challenges with sensitivity, selectivity, and signal interference.

Purpose of the Study:

To develop a novel and highly sensitive surface-enhanced Raman scattering (SERS) based biosensor for cytochrome c detection.
To utilize target-binding-induced conformational changes of a signal transduction probe (STP) for signal amplification.
To optimize STP surface density for enhanced sensitivity and reduced non-specific signal interference.

Main Methods:

Immobilization of a SERS-active signal transduction probe (STP) labeled with carboxy-X-rhodamine (ROX) on a substrate.

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Utilizing aptamers to form a rigid triplex structure that initially suppresses SERS amplification.

Monitoring changes in ROX SERS intensity upon cytochrome c binding, which induces conformational changes and signal enhancement.

Main Results:

Achieved a low detection limit of 2 nM (10 fmol in 5 μL sample solution) for cytochrome c.
Demonstrated excellent selectivity for cytochrome c over interfering proteins like hemoglobin and immunoglobulin G.
Optimized STP surface density to effectively shield background signals from protein adsorption.

Conclusions:

The developed STP-based SERS method offers a sensitive and selective approach for cytochrome c detection.
The strategy of using target-induced conformational changes for SERS signal transduction is effective.
This generalizable strategy can be extended to develop aptamer or DNA-based sensors for other analytes.