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Enzyme microgels in packed-bed bioreactors with downstream amperometric detection using microfabricated
A Guiseppi-Elie1, N F Sheppard, S Brahim
1Department of Chemical Engineering and Center for Bioelectronics, Biosensors and Biochips (C3B), Virginia Commonwealth University, P.O. Box 843028, 601 West Main Street, Richmond, VA 23284-3028, USA. guiseppi@vcu.edu
Biotechnology and Bioengineering
|October 23, 2001
Summary
pH-responsive hydrogels effectively immobilize glucose oxidase (GOx) and glutamate oxidase (GlutOx) for biosensing. These enzyme-immobilized hydrogels demonstrate stability and high performance in flow injection systems for quantifying glucose and MSG.
Area of Science:
- Biomaterials Science
- Analytical Chemistry
- Enzyme Immobilization
Background:
- Enzyme immobilization is crucial for developing stable and reusable biosensors.
- pH-responsive hydrogels offer tunable environments for enzyme activity and stability.
- Flow injection analysis (FIA) systems require robust bioactive components for continuous monitoring.
Purpose of the Study:
- To develop pH-responsive hydrogels for immobilizing glucose oxidase (GOx) and glutamate oxidase (GlutOx).
- To integrate these enzyme-immobilized hydrogels into flow injection systems for analyte quantification.
- To evaluate the performance and stability of the developed biosensing systems.
Main Methods:
- Spherical hydrogel beads were synthesized using inverse suspension polymerization.
- Enzymes (GOx and GlutOx) were immobilized via physical entrapment or covalent attachment.
- Packed-bed bioreactors containing the hydrogels were incorporated into flow injection amperometric systems.
Main Results:
- The GOx-immobilized system showed linearity for glucose detection from 1.8-280 mg/dL with a 1.4 mg/dL detection limit.
- The GlutOx-immobilized system achieved linearity up to 135 mg/dL for MSG with a 3.38 mg/dL detection limit.
- Physically entrapped enzymes retained over 60% activity after one year of storage, demonstrating excellent stability.
Conclusions:
- pH-responsive hydrogels are effective matrices for immobilizing GOx and GlutOx for biosensing applications.
- The developed flow injection systems offer high sensitivity, good precision, and excellent long-term stability.
- These immobilized enzyme systems show promise for the reliable quantification of glucose and MSG in various samples.