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Biosensor based on glucose dehydrogenase and detecting method

A glucose dehydrogenase, biosensor technology, applied in instruments, measuring devices, scientific instruments, etc., to achieve good linearity

Inactive Publication Date: 2017-08-18
TAIZHOU E LINKCARE MEDITECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Aiming at the problems and defects of the above-mentioned prior art, the object of the present invention is to utilize the advantages of good specificity and no oxygen content interference of flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase, combined with ruthenium complexation Based on the advantages of good stability and low background noise, to solve the problem of the compatibility of the two reactions, develop a high-performance biosensor based on glucose dehydrogenase, and use the biosensor to detect the glucose content in the sample sensitively.

Method used

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  • Biosensor based on glucose dehydrogenase and detecting method
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  • Biosensor based on glucose dehydrogenase and detecting method

Examples

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Embodiment 1

[0053] The biosensor structure of Example 1 is the same as that of Comparative Example 1, except that the components of the detection reagent are different. The components of the aqueous solution of the detection reagent are as follows: 100 mM sodium phosphate buffer (pH 7.0), 1.2% (w / v) methylcellulose (Sigma M6385), 0.5% (w / v) polyethylene glycol 4000 (Sigma 95904), 0.5% (w / v) Triton X-100 (Aladdin T109026), 0.5% (w / v) BSA (Aladdin A104912), 1.86 % (w / v) hexaammine ruthenium trichloride (CMI 8015), 0.16% (w / v) 2,6-dichlorophenol indophenol sodium (DCPIP, J&K 249712), 2000 U / mL flavin adenine Dinucleotide (FAD)-dependent glucose dehydrogenase (BBI Solutions GLD-3). The detection reagent aqueous solution was dispensed and fixed on the electrode system of the biosensor test piece using a dispenser, and each biosensor test piece was dispensed with 1.0uL detection reagent aqueous solution, and then placed in a 60°C oven for 10 minutes to dry.

Embodiment 2

[0055] The biosensor structure of Example 2 is the same as that of Comparative Example 1, except that the components of the detection reagent are different. The components of the aqueous solution of the detection reagent are as follows: 100 mM sodium phosphate buffer (pH 7.0), 1.2% (w / v) methylcellulose (Sigma M6385), 0.5% (w / v) polyethylene glycol 4000 (Sigma 95904), 0.5% (w / v) Triton X-100 (Aladdin T109026), 0.5% (w / v) BSA (Aladdin A104912), 1.86 % (w / v) hexaamine ruthenium trichloride (CMI 8015), 0.08% (w / v) 2,6-dichlorophenol indophenol sodium (DCPIP, J&K 249712), 2000 U / mL flavin adenine Dinucleotide (FAD)-dependent glucose dehydrogenase (BBI Solutions GLD-3). The detection reagent aqueous solution was dispensed and fixed on the electrode system of the biosensor test piece using a dispenser, and each biosensor test piece was dispensed with 1.0uL detection reagent aqueous solution, and then placed in a 60°C oven for 10 minutes to dry.

Embodiment 3

[0057] The structure of the biosensor of Example 3 is the same as that of Comparative Example 1, except that the components of the detection reagent are different. The components of the aqueous solution of the detection reagent are as follows: 100 mM sodium phosphate buffer (pH 7.0), 1.2% (w / v) methylcellulose (Sigma M6385), 0.5% (w / v) polyethylene glycol 4000 (Sigma 95904), 0.5% (w / v) Triton X-100 (Aladdin T109026), 0.5% (w / v) BSA (Aladdin A104912), 1.24 % (w / v) hexaammine ruthenium trichloride (CMI 8015), 0.16% (w / v) 2,6-dichlorophenol indophenol sodium (DCPIP, J&K 249712), 2000 U / mL flavin adenine Dinucleotide (FAD)-dependent glucose dehydrogenase (BBI Solutions GLD-3). The detection reagent aqueous solution was dispensed and fixed on the electrode system of the biosensor test piece using a dispenser, and each biosensor test piece was dispensed with 1.0uL detection reagent aqueous solution, and then placed in a 60°C oven for 10 minutes to dry.

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Abstract

The invention relates to the field of a biological detection instrument, and particularly relates to a biosensor based on glucose dehydrogenase. The biosensor is composed of an insulation substrate, an electrode system, a detecting reagent, a middle interlayer, a covering layer, a reaction room and a sample injection channel; the reaction room is provided with air holes. Enzyme in the detection reagent is flavin adenine dinucleotide dependent form glucose dehydrogenase; the electronic medium is commonly formed by ruthenium complex and 2, 6-dichlorophen indophenol. The invention further provides a detecting method of the biosensor. The biosensor solves the problem that the pure ruthenium complex is very hard to accept electronic from the flavin adenine dinucleotide dependent form glucose dehydrogenase, thus the biosensor has the advantages of good reaction sensitivity and linearity.

Description

technical field [0001] The invention relates to the field of biological detection instruments, in particular to a biosensor and detection method based on flavin adenine dinucleotide (FAD)-dependent glucose dehydrogenase. Background technique [0002] Biosensor (biosensor) is an instrument that uses biologically active substances to selectively identify target substances and convert their concentration or content into electrical signals for detection. It uses immobilized biosensitive materials as recognition elements (including enzymes, antibodies, Antigens, microorganisms, cells, tissues, nucleic acids and other biologically active substances), appropriate physical and chemical transducers (such as oxygen electrodes, photosensitive tubes, field effect tubes, piezoelectric crystals, etc.) and signal amplification devices constitute analysis tools or systems. [0003] Electrochemical biosensors have been widely used in the field of in vitro diagnostics. These instrument system...

Claims

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Application Information

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IPC IPC(8): G01N27/327
CPCG01N27/3272G01N27/3277
Inventor 章子毅李红周嘉强郑芬萍李霖吴建平王天星
Owner TAIZHOU E LINKCARE MEDITECH CO LTD
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