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An electrochemical biosensor for detecting glutathione and a preparing method thereof

A biosensor, glutathione technology, applied in the direction of material electrochemical variables, scientific instruments, instruments, etc., to achieve the effect of strong affinity, wide application prospects, and sensitive detection

Inactive Publication Date: 2015-01-07
SHANGHAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods have their own advantages in detection sensitivity, selectivity, detection time and stability, but at the same time, they also have disadvantages such as cumbersome operation and expensive equipment.

Method used

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  • An electrochemical biosensor for detecting glutathione and a preparing method thereof
  • An electrochemical biosensor for detecting glutathione and a preparing method thereof
  • An electrochemical biosensor for detecting glutathione and a preparing method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Preparation of double-stranded DNA modified gold electrode

[0028] Drop 20 μL of piranha solution (concentrated sulfuric acid: hydrogen peroxide = 3:1) on the surface of the gold electrode to be treated and react for 2 minutes, rinse with ultrapure water, and blow dry with nitrogen. After polishing the gold electrode on 5000-mesh sandpaper for 5 minutes, it was polished to the mirror surface on the silk containing alumina (grain size of 1 μm, 0.3 μm, 0.05 μm) mortar, and then sonicated in ethanol and ultrapure water. 2 minutes to remove impurities. Place the processed gold electrode at 0.5 M H 2 SO 4 Cyclic voltammetry scan. Set the scan voltage range from 0 to 1.6 V, scan speed 100 mV / s, set 30 cycles to sweep until the cyclic voltammetry curve is stable, and blow dry with nitrogen to obtain a clean surface-treated bare gold electrode, which can be used for modification of sulfhydryl DNA. Immerse the gold electrode in a DNA fixation buffer solution (10 mM Tr...

Embodiment 2

[0029] Example 2: Synthesis and electrochemical quantitative characterization of copper nanoclusters

[0030] Take 40 μL 10 mM ascorbic acid solution and 320 μL reaction buffer (20 mM MOPS, 300 mM NaCl and 2 mM MgCl 2 , PH 7.5) mix, then add 40 μL of copper ion solution of different concentrations to the system, and let stand for 20 minutes. Put the double-stranded DNA modified gold electrode into the above mixed solution and react for 30 minutes at room temperature. After the reaction, the electrode was rinsed with ultrapure water and dried with nitrogen to remove excess copper ions adsorbed on the electrode surface. Subsequently, the electrode was immersed in 200 μL of 0.1 M hydrochloric acid solution to dissolve the copper nanoclusters synthesized on the electrode surface. After 2 hours, the above solution was mixed with 4.8 mL of 0.5 M NaAc-HAc buffer and used as an electrolyte solution for electrochemical detection.

[0031] The relevant nucleic acid sequence is as follows: ...

Embodiment 3

[0036] Example 3: Detection of different concentrations of glutathione

[0037] Take 40 μL 10 mM ascorbic acid solution and 320 μL reaction buffer (20 mM MOPS, 300 mM NaCl and 2 mM MgCl 2 , PH 7.5), and then add 40 μL of 10 μM copper ions and different concentrations of glutathione (0 nM, 1 nM, 5 nM, 10 nM, 100 nM, 1 μM and 10 μM) to the system. Test the mixed solution and react at room temperature for 20 minutes. Subsequently, the double-stranded DNA-modified gold electrode was put into the above-mentioned mixed solution, reacted at room temperature for 30 minutes, and then electrochemical detection was performed. The specific detection steps were the same as those in the second embodiment.

[0038] figure 2 Curve a shows the DPV spectrum obtained by scanning the double-stranded DNA modified gold electrode in a reaction system containing only ascorbic acid and copper ions for a period of time and after a certain treatment. It can be seen that the double-stranded DNA molecules im...

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Abstract

The invention relates to an electrochemical biosensor for detecting glutathione and a preparing method thereof. The biosensor is a three-electrode system sensor, wherein the counter electrode is a platinum electrode, the reference electrode is a saturated calomel electrode, and the working electrode is a gold electrode. The electrochemical biosensor and the method are characterized in that: the gold electrode is modified with double-stranded DNA capable of being adopted as a copper nanocluster synthesis template. According to the electrochemical biosensor and the preparing method, high-affinity bonding of the glutathione and a copper ion is utilized, synthesis of copper nanoclusters on surfaces of the electrodes is inhibited, and indirect detection of the glutathione is achieved by performing electrochemical quantitative characterization on the copper nanoclusters. The linear range of detection of the glutathione is 1-1000 nM, and the detection limit is about 0.42 nM. The electrochemical biosensor and the method have advantages of simple operation, low cost, using convenience, high selectivity, and the like, so that the electrochemical biosensor and the method have large potential application value in the fields of biochemical research, clinical analysis, and other fields.

Description

technical field [0001] The invention relates to a novel electrochemical biosensor and a preparation method thereof, in particular to an electrochemical biosensor for detecting glutathione and a preparation method thereof. Background of the invention [0002] Glutathione is the most abundant small molecular weight non-protein sulfhydryl compound in cells, which is formed by condensation of glutamic acid, cysteine ​​and glycine through peptide bonds. Glutathione is a regulator of sulfhydryl groups and sulfides in the body, and plays an important role in maintaining the reduction state of protein sulfhydryl groups and enzyme activity, anti-oxidation, and maintaining the balance of the redox environment in biological organisms. Several studies in recent years have shown that changes in the concentration of glutathione in the body are related to the occurrence and development of many diseases such as Alzheimer's disease, Parkinson's syndrome, diabetes, and atherosclerosis; and t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/403G01N27/48
Inventor 李根喜曹亚虞加翠张慧慧陈伟伟韩鹏
Owner SHANGHAI UNIV
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