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Published on: December 1, 2015
Dithiobissuccinimidyl propionate self assembled monolayer based cholesterol biosensor
Sunil K Arya1, Pratibha Pandey, Surinder P Singh
1Biomolecular Electronics & Conducting Polymer Research Group, National Physical Laboratory, Dr. K. S. Krishnan Marg, New Delhi-110012, India.
The Analyst
|September 26, 2007
Summary
Researchers developed a novel biosensor using dithiobissuccinimidyl propionate (DTSP) to immobilize cholesterol enzymes on a gold surface. This enhanced biosensor demonstrates high enzyme affinity and stability for cholesterol detection.
Area of Science:
- Biotechnology
- Biosensor Development
- Electrochemistry
Background:
- Biosensors are crucial for detecting biomarkers like cholesterol.
- Enzyme immobilization techniques impact biosensor performance and stability.
- Gold surfaces offer excellent platforms for biosensor fabrication.
Purpose of the Study:
- To develop a stable and sensitive biosensor for cholesterol detection.
- To covalently immobilize cholesterol oxidase (ChOx) and cholesterol esterase (ChEt) onto a gold surface.
- To characterize the performance of the developed ChOx-ChEt/DTSP/Au bio-electrode.
Main Methods:
- Fabrication of a self-assembled monolayer (SAM) using dithiobissuccinimidyl propionate (DTSP) on a gold (Au) surface.
- Covalent immobilization of cholesterol oxidase (ChOx) and cholesterol esterase (ChEt) onto the DTSP/Au surface.
- Characterization using electrochemical impedance spectroscopy and cyclic voltammetry.
- Enzymatic assay using differential pulse voltammetry (DPV) to determine cholesterol oleate concentration.
Main Results:
- The ChOx-ChEt/DTSP/Au bio-electrode exhibited enhanced enzyme affinity with a Michaelis-Menten constant (Km) of 0.95 mM.
- The biosensor demonstrated linearity for cholesterol oleate detection in the range of 50 to 400 mg dl(-1).
- The developed bio-electrode showed a shelf-life exceeding 50 days when stored at 4°C, with a high correlation coefficient (0.9973).
Conclusions:
- The DTSP-based SAM provides an effective platform for covalent enzyme immobilization, leading to a highly sensitive and stable cholesterol biosensor.
- The developed ChOx-ChEt/DTSP/Au bio-electrode shows significant potential for accurate and reliable cholesterol monitoring.
- This approach offers a promising strategy for the development of advanced electrochemical biosensing platforms.
