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Electrochemical biosensor for detecting intracellular hydrogen peroxide on line based on paper chip in combination with screen printing electrode

A technology of screen-printed electrodes and biosensors, applied in the field of detection, can solve the problems of large sample volume, complicated operation, unfavorable online monitoring of cell levels, etc., and achieve the effect of strong sensitivity and high specificity

Active Publication Date: 2019-01-08
FUJIAN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, traditional electrodes have disadvantages such as large amount of sample required, complex operation, difficulty in integration and miniaturization, and are not conducive to online monitoring at the cellular level.

Method used

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  • Electrochemical biosensor for detecting intracellular hydrogen peroxide on line based on paper chip in combination with screen printing electrode
  • Electrochemical biosensor for detecting intracellular hydrogen peroxide on line based on paper chip in combination with screen printing electrode
  • Electrochemical biosensor for detecting intracellular hydrogen peroxide on line based on paper chip in combination with screen printing electrode

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] (1) Preparation of nafion / PtNPs / MWCNTs / SPCE electrode

[0034] Clean the screen-printed carbon electrode three times with deionized water, dry it with nitrogen gas, drop-coat the multi-walled carbon nanotube material dispersion on the surface of the working electrode of the screen-printed carbon electrode, let it dry naturally, and then drop 2 mmol / L chloroplatinic acid solution and 0.5 mol / L sulfuric acid solution were used to deposit platinum nanoparticles on the surface of the working electrode of the screen-printed carbon electrode by cyclic voltammetry. Finally, add nafion dropwise and let it dry naturally to form a nafion film, that is, a nano-platinum / multi-walled carbon nanotube modified screen-printed electrode (nafion / PtNPs / MWCNTs / SPCE) ( figure 1 No. 7 in: as a working electrode). Such as figure 1 As shown, a reference electrode 6, a working electrode 7 and a counter electrode 8 are integrated in the screen-printed carbon electrode 4, and a reference elect...

Embodiment 2

[0043] An electrochemical biosensor based on a paper chip combined with a screen-printed electrode for online detection of intracellular hydrogen peroxide is used for online monitoring of PC12 cells releasing hydrogen peroxide. The detection steps are as follows:

[0044] When the PC12 cells (adrenal pheochromocytoma cells) grow to 80% of the culture bottle, and the growth state is observed to be good, pour out the culture medium, rinse with phosphate buffer saline (PBS) three times, and then use an appropriate amount of The PBS solution (0.1 mol / L, pH7.4) contacted the cells, placed in the incubator for 5 minutes, collected the PBS buffer solution contacting the PC12 cells, and used the solution to prepare a series of hydrogen peroxide solutions, using time-current (i-t) method, recording the current response value of different concentrations of hydrogen peroxide at a voltage of -0.3 V and a time of 50 s.

[0045] Depend on image 3 It can be seen that the current response s...

Embodiment 3

[0047] An electrochemical biosensor based on a paper chip combined with a screen-printed electrode for on-line detection of intracellular hydrogen peroxide is used for on-line monitoring of AD models. The steps are as follows:

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Abstract

The invention discloses an electrochemical biosensor for detecting intracellular hydrogen peroxide on line based on a paper chip in combination with a screen printing electrode. The prepared nano-platinum / multi-walled carbon nanotube material is modified on the surface of the screen printing electrode and organically combined with a self-made integrated cell culture platform-paper-based chip to manufacture a biosensor for monitoring PC12 cells (adrenal pheochromocytoma cells) that releases hydrogen peroxide on line. The biosensor is connected with an electrochemical workstation to monitor theactivities of PC12 cells and Alzheimer's disease (AD) model cells, has good specificity and high sensitivity, and provides a novel method for research on cell activity monitoring and other related fields.

Description

technical field [0001] The invention relates to an electrochemical biosensor for online detection of intracellular hydrogen peroxide based on a paper chip combined with a screen printing electrode, belonging to the detection technical field. The paper chip can be used for detecting hydrogen peroxide in PC12 cells and AD model cells. Background technique [0002] The most common age-related neurodegenerative disease is Alzheimer's disease, the pathogenic mechanism of which involves the aggregation and deposition of β-amyloid. Studies have found that the accumulation and overload of β-amyloid (Aβ) can cause oxidative stress, promote the influx of intracellular calcium ions, lead to the depolarization of mitochondrial membranes, and increase the expression level of intracellular hydrogen peroxide. Therefore, the realization of the content detection of hydrogen peroxide molecular compounds at the cellular level is of great significance for understanding the pathological mechanis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N27/30
CPCG01N27/30G01N27/308
Inventor 刘爱林刘萌萌李珊红刘辉雷云彭花萍陈伟林新华
Owner FUJIAN MEDICAL UNIV
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