Non-Destructive Monitoring via Electrochemical NADH Detection in Murine Cells

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Cell Biology

Background:

• Nicotinamide adenine dinucleotide (NADH) is a crucial cofactor in cellular metabolism, essential for redox reactions.
• Accurate detection of NADH in living cells presents significant analytical challenges.
• Existing methods for NADH detection are often complex or lack sensitivity for in vivo applications.

Purpose of the Study:

• To develop a sensitive and selective electrochemical sensor for the one-step detection of NADH.
• To utilize a surface-modified screen-printed electrode (SPE) for enhanced NADH monitoring.
• To demonstrate the sensor's utility in assessing cell viability through NADH detection.

Main Methods:

• Fabrication of a screen-printed electrode (SPE) modified with a redox-active monolayer (4'-mercapto-N-phenlyquinone diamine, NPQD) via self-assembled monolayer (SAM) of 4-aminothiophenol (4-ATP).
• Electrocatalytic detection of NADH using the modified SPE in cell culture media.
• Validation of sensor performance, including limit of detection (LOD) and sensitivity.

Main Results:

• The developed NADH sensor achieved a low limit of detection (LOD) of 0.49 μM.
• The sensor exhibited high sensitivity (0.0076 ± 0.0006 μA/μM) and selectivity in cell culture media.
• Demonstrated successful application in monitoring cell viability by detecting NADH in cell culture supernatants.

Conclusions:

• The novel electrochemical sensor provides a reliable and efficient method for NADH detection in biological samples.
• This technology offers a promising tool for real-time monitoring of cellular metabolic status and cell viability.
• The one-step detection approach simplifies NADH analysis, paving the way for broader applications in cell biology research.