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Automated Quantitative Enzyme Biosensing in 24-Well Microplates.

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Summary

This study introduces enzyme-modified pencil leads as novel electrochemical biosensors for automated glucose estimation. The cost-efficient robotic system offers accurate, hands-off quantification in microplates for various analyses.

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Area of Science:

Electrochemistry
Biosensors
Analytical Chemistry

Background:

Accurate glucose estimation is crucial for clinical and research applications.
Existing methods can be labor-intensive and prone to operator error.
Development of cost-effective and automated biosensing platforms is needed.

Purpose of the Study:

To develop a novel, automated system for glucose estimation using enzyme-modified pencil leads as electrochemical biosensors.
To evaluate the performance of this system in model and real samples.
To demonstrate a cost-efficient and labor-saving approach to quantitative biosensing.

Main Methods:

Enzyme (glucose oxidase) immobilization onto electrochemically modified graphite pencil leads (PL).
Automated amperometric sensing using a robotic system with computer-controlled electrodes in microtiter plates.
Assay validation using standard solutions and human serum samples in calibration curve and standard addition modes.

Main Results:

The enzyme-modified PL biosensors demonstrated a stable and linear response for glucose detection (0.1-8 mM).
A low detection limit of 0.05 ± 0.01 mM was achieved.
Accurate and reproducible glucose quantification (>99% recovery) was obtained in both standard solutions and human serum samples.
The automated system allowed for hands-off evaluation of 4 or 20 samples per run.

Conclusions:

Enzyme-modified pencil leads serve as effective and inexpensive electrochemical biosensors for glucose quantification.
The developed robotic system provides a convenient, labor- and cost-efficient, and accurate method for automated biosensing.
This methodology holds potential for repetitive sample analysis in academic research, clinical diagnostics, and environmental/pharmaceutical/biotechnological applications.