Simultaneous Determination of Glucose and Analogous Disaccharides by Dual-Electrode Enzyme Sensor System

Zhang, Xian-En

doi:10.1007/978-1-4615-4181-3_14

“…Particularly, determination of glucose in blood plays a crucial role in the clinical diagnosis and treatment therapy of diabetes. The amperometric glucose sensor is one of the most important enzyme-based biosensors (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14) due to their simple, sensitive and inexpensive merits. In amperometric glucose sensors, solid electrode surface is chemically modified by a sensing layer containing redox centers of glucose oxidase (GOD), or both GOD and electron mediators for glucose determination (15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Porous carbon composite/enzyme glucose microsensor

Li

¹

,

S²

2004

View full text Add to dashboard Cite

An enzyme glucose microsensor using a glucose oxidase-immobilized porous carbon/Teflon composite microelectrode was developed. The microsensor was fabricated by etching a platinum microelectrode (platinum, radius of 25 and 50 micrometers) in hot aqua regia to create a cavity at the tip and then packing a porous carbon/Teflon composite, which was made from acetylene black and Teflon emulsion, into the cavity. Nafion was impregnated into the inner surface of porous carbon/Teflon composite electrode following immobilization of Os(bpy)3+2/+3 as electron transfer mediators. The loading amount of Os(bpy)3+2/+3 in the Nafion/porous carbon/Teflon composite electrode was found to be 7.0x10(-8) mole cm(-2), which is much higher than that in polymer modified electrodes reported in literatures. The microsensor was further dipped overnight in buffer solution containing glucose oxidase for enzyme modification. With both glucose oxidase and mediators in the porous carbon/Teflon composite surface, the sensor performance was evaluated in buffer solutions containing different glucose concentrations and serum samples for glucose determination. The microsensor showed directly electrochemical glucose oxidation on the Os(bpy)3+2/+3 impregnated enzyme/porous carbon/Teflon composite surface with linear response over concentration range of 0-15 mM and Machaelis behavior. Reliability and reproducibility were conducted in serum samples and glucose buffer solution, and the results demonstrated there was no significant decrease of amperometric response in air-saturated solution for one month. The sensor demonstrated potential in clinical diagnostic applications.

show abstract

“…Failure of the control results in poor linearity, sensitivity, and reproducibility (also known as interchangeability); the last one is extremely important in the manufacture of disposable biosensors. Many investigators tried to optimize the enzyme ratio in the membrane simply by adjusting the enzyme loadings ( − ), but the results were not always satisfying because the immobilization efficiency of the enzymes may differ from each other under the same immobilization conditions. On the basis of the studies of protein molecular structure, here we proposed a fusion enzyme method by gene manipulation to solve the problems.…”

Section: Introductionmentioning

confidence: 99%

“…Sequence biosensors, which involve two or more enzymes for sequential catalysis, are widely used in the determination of biochemical substances, such as disaccharides ( − ), starch (), inosine (), total cholesterol (), etc. Upon review of the abstract books of the World Congress on Biosensors of the past decade, it is found that about 30% of the reported biosensors are sequence biosensors.…”

Section: Introductionmentioning

confidence: 99%

Construction of a Fusion Enzyme System by Gene Splicing as a New Molecular Recognition Element for a Sequence Biosensor

Zhou

¹

,

Zhang

²

,

Liu

³

et al. 2001

Self Cite

View full text Add to dashboard Cite

A bifunctional fusion enzyme system constructed by gene splicing is proposed as a new model to develop sequence biosensors, taking maltose biosensor as an example. The cDNA fragment of Aspergillus niger glucoamylase (E.C 3.2.1.3, GA) was fused to the 3' end of Aspergillus niger glucose oxidase (E.C 1.1.3.4, GOD) gene with the insertion of a flexible linker peptide [-(Ser-Gly)5-] coding sequence. The fusion gene was cloned into the vector pPIC9 and expressed in Pichia pastoris GS115 under the control of the AOX1 promoter. It was found that a bifunctional hybrid protein with a molecular weight of 430 kDa was secreted after induction with methanol. The fusion enzyme GOD-(Ser-Gly)5-GA (GLG) was purified using Q Sepharose Fast Flow ion-exchange chromatography. Kinetic analysis demonstrated that GLG retained the typical kinetic properties of both GA and GOD. After being immobilized on an aminosilanized glass slide through covalent bonding by glutaraldehyde, GLG showed much higher sequential catalytic efficiency than the mixture of separately expressed GA and GOD (GA/GOD). Maltose biosensors were fabricated with GLG and GA/GOD, respectively. The performance characteristics of the maltose biosensor with respect to reproducibility, signal level, and linearity were effectively improved by using the fusion enzyme. Our findings offer a basis for the development of other sequence biosensors.

show abstract

“…Particularly, determination of glucose in blood plays a crucial role in the clinical diagnosis and treatment therapy of diabetes. The amperometric glucose sensor is one of the most important enzyme-based biosensors (1)(2)(3)(4)(5)(6)(7)(8)(9)(10)(11)(12)(13)(14) due to their simple, sensitive and inexpensive merits. In amperometric glucose sensors, solid electrode surface is chemically modified by a sensing layer containing redox centers of glucose oxidase (GOD), or both GOD and electron mediators for glucose determination (15)(16)(17).…”

Section: Introductionmentioning

confidence: 99%

Porous carbon composite/enzyme glucose microsensor

2004

View full text Add to dashboard Cite

An enzyme glucose microsensor using a glucose oxidase-immobilized porous carbon/Teflon composite microelectrode was developed. The microsensor was fabricated by etching a platinum microelectrode (platinum, radius of 25 and 50 micrometers) in hot aqua regia to create a cavity at the tip and then packing a porous carbon/Teflon composite, which was made from acetylene black and Teflon emulsion, into the cavity. Nafion was impregnated into the inner surface of porous carbon/Teflon composite electrode following immobilization of Os(bpy)3+2/+3 as electron transfer mediators. The loading amount of Os(bpy)3+2/+3 in the Nafion/porous carbon/Teflon composite electrode was found to be 7.0x10(-8) mole cm(-2), which is much higher than that in polymer modified electrodes reported in literatures. The microsensor was further dipped overnight in buffer solution containing glucose oxidase for enzyme modification. With both glucose oxidase and mediators in the porous carbon/Teflon composite surface, the sensor performance was evaluated in buffer solutions containing different glucose concentrations and serum samples for glucose determination. The microsensor showed directly electrochemical glucose oxidation on the Os(bpy)3+2/+3 impregnated enzyme/porous carbon/Teflon composite surface with linear response over concentration range of 0-15 mM and Machaelis behavior. Reliability and reproducibility were conducted in serum samples and glucose buffer solution, and the results demonstrated there was no significant decrease of amperometric response in air-saturated solution for one month. The sensor demonstrated potential in clinical diagnostic applications.

show abstract

Simultaneous Determination of Glucose and Analogous Disaccharides by Dual-Electrode Enzyme Sensor System

Cited by 3 publications

References 17 publications

Porous carbon composite/enzyme glucose microsensor

Porous carbon composite/enzyme glucose microsensor

Construction of a Fusion Enzyme System by Gene Splicing as a New Molecular Recognition Element for a Sequence Biosensor

Porous carbon composite/enzyme glucose microsensor

Contact Info

Product

Resources

About