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Ultrasensitive SERS Profiling of Intracellular Hydrogen Peroxide Release Based on Enzymatic Amplification and
Jindan Wu1, Kaixin Chen1, Junming Pan1
1Key Laboratory of Biomaterials of Guangdong Higher Education Institutes, Department of Biomedical Engineering, Jinan University, Guangzhou 510632, China.
Analytical Chemistry
|December 5, 2024
Summary
This study introduces an ultrasensitive surface-enhanced Raman scattering (SERS) sensor for detecting cellular oxidative stress. The novel sensor utilizes enzymatic amplification and a silent-range Raman fingerprint to achieve high sensitivity and low background noise for biological applications.
Area of Science:
- Analytical Chemistry
- Biomedical Sensing
- Spectroscopy
Background:
- Surface-enhanced Raman scattering (SERS) offers high sensitivity for biomarker detection.
- Severe background noise in SERS limits practical applications, especially in biological systems.
- Accurate monitoring of cellular oxidative stress is crucial for understanding disease mechanisms.
Purpose of the Study:
- To develop an ultrasensitive SERS sensor for determining cellular oxidative stress.
- To overcome limitations of background noise in SERS through a novel sensing strategy.
- To enable sensitive and reliable monitoring of hydrogen peroxide (H₂O₂) levels in biological samples.
Main Methods:
- Utilized horseradish peroxidase (HRP) for H₂O₂-induced enzymatic amplification.
- Employed a coupling reaction between 4-hydroxythiophenol (4-MTP) and phenol-d5 to form a SERS signal.
- Leveraged the Raman-silent fingerprint of phenol-d5 at 2125 cm⁻¹ to minimize background noise.
- Developed a SERS substrate with Au-S bonding for stable signal generation.
Main Results:
- Achieved ultrasensitive determination of H₂O₂ with a broad linear range (5 × 10⁻⁹ to 1 × 10⁻³ M).
- Demonstrated a low limit of detection for H₂O₂ as 1.6 nM.
- Successfully profiled dynamic changes in intracellular H₂O₂ release.
- Distinguished between cancerous and healthy cells based on oxidative stress levels.
- Identified varying susceptibility to oxidative stimulation in different cell populations.
Conclusions:
- The developed SERS sensor provides a sensitive and anti-interference method for H₂O₂ detection.
- The combination of enzymatic amplification and silent-range fingerprinting significantly enhances SERS performance.
- This approach is effective for monitoring endogenous oxidative behavior in living cells.
- The study demonstrates the potential of SERS for diagnosing and understanding oxidative stress-related diseases.
