Reversible Two-Enzyme Coimmobilization on pH-Responsive Imprinted Monolith for Glucose Detection

Area of Science:

• Biomedical Engineering
• Analytical Chemistry
• Materials Science

Background:

• Development of sensitive and selective glucose biosensors is crucial for diabetes management.
• Existing biosensors often face challenges with stability and reusability.
• Enzyme immobilization techniques are key to improving biosensor performance.

Purpose of the Study:

• To develop a novel, reusable enzymatic glucose biosensor.
• To utilize a pH-responsive imprinted polymer for enzyme co-immobilization.
• To enhance glucose detection selectivity and specificity.

Main Methods:

• Co-immobilization of horseradish peroxidase (HRP) and glucose oxidase (GOx) on a pH-responsive imprinted monolith.
• Enzyme conjugation via biotin-streptavidin affinity.
• Surface-initiated atom transfer radical polymerization for polymer grafting.
• pH-modulated enzyme stripping and re-immobilization for reusability.

Main Results:

• The biosensor exhibited a linear glucose detection range of 0.11–38.85 mmol/L.
• A low limit of detection of 0.03 mmol/L was achieved.
• High selectivity and specificity were confirmed in human serum samples, outperforming traditional methods.
• A relative standard deviation of 3.7% indicated good reproducibility.

Conclusions:

• A novel, pH-responsive imprinted polymer-based glucose biosensor was successfully developed.
• The co-immobilization strategy and pH-modulated reusability offer significant advantages.
• This enzymatic sensing system shows great potential for clinical glucose monitoring.