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Preparation method and application of hydrogel electrode

A hydrogel and electrode technology, which is applied in the preparation of hydrogel electrodes and the field of electrochemical detection of mitochondrial DNA mutations in ovarian cancer, can solve the problems of inactivation of biologically active molecules, difficulty in clinical application, complex structure, etc., and achieve simple preparation methods , Improving hybridization efficiency and low cost

Active Publication Date: 2014-10-08
XIAMEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[6-8] However, the graphene-based electrochemical biosensors reported in the current literature are not only complicated in structure, but also the bioactive molecules immobilized on the surface are inactivated due to long-term storage, making it difficult to really go to clinical application.

Method used

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  • Preparation method and application of hydrogel electrode
  • Preparation method and application of hydrogel electrode
  • Preparation method and application of hydrogel electrode

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Accurately weigh 2g of natural graphite powder and 1g of sodium nitrate into a round-bottomed flask, and mix evenly with 46mL of concentrated sulfuric acid in an ice bath; then slowly add 6g of potassium permanganate to the mixture to keep the mixture at a low temperature Stir and react at 20°C for 2h, then transfer the mixture to an oil bath at 35±5°C for 30min, the reaction system is a brown viscous liquid; then slowly add 92mL deionized water to the mixture , and raise the temperature to 95±5°C to continue the reaction for 3 hours, the mixture turns from brown to bright yellow, and finally add 400mL of pure water to terminate the reaction, and at the same time add 6mL of 30% H 2 o 2 The solution neutralizes unreacted potassium permanganate. After the above solution was cooled to room temperature, suction filtration was performed, and the filter cake was repeatedly washed with 100 mL aqueous hydrochloric acid (1:10) and a large amount of pure water to remove residual...

Embodiment 2

[0045] Accurately draw an equal volume of graphene oxide aqueous solution (6mg / mL) and protamine DNA aqueous solution (10mg / mL) and mix them evenly in a beaker; take a clean 2mL centrifuge tube, and punch a small hole with a diameter of about 1mm at the bottom , wrap the bottom tightly with plastic wrap for later use; use a micropipette to absorb an appropriate amount of graphene oxide and protamine DNA mixed solution, and add it vertically and slowly into the centrifuge tube to ensure that the liquid level in the tube is as horizontal as possible and the tube wall is clean and free of contamination. Residual droplets; start the heating mixer and preheat to 95°C; carefully move the centrifuge tube containing the mixed solution of graphene oxide and protamine DNA into the heating hole of the mixer, and take it out after heating for 10 minutes. The mixed solution solidifies to form graphene oxide / DNA hydrogel. The hydrogel was sealed and stored at 4°C. Optical photographs and s...

Embodiment 3

[0049] Draw an appropriate amount of 1% polyethyleneimine (PEI) solution and add it to the top of the hydrogel in the graphene oxide / DNA hydrogel electrode. After standing for 30 minutes, discard the PEI solution in the electrode and clear the water to obtain PEI-modified Graphene oxide / DNA hydrogel electrodes. Add the oligonucleotide probe solution to the top of the hydrogel in the PEI-modified graphene oxide / DNA hydrogel electrode, incubate at 4°C for 30 min, discard the oligonucleotide probe solution in the electrode, and Wash with PBS and ultrapure water to obtain a PEI-modified graphene oxide / DNA hydrogel electrode immobilized with oligonucleotide probes. Another graphene oxide / DNA hydrogel electrode not modified by PEI was taken and incubated with the same method and conditions to obtain a graphene oxide / DNA hydrogel electrode immobilized with oligonucleotide probes. A graphene oxide / DNA hydrogel electrode was used as the working electrode, and a platinum wire electrode...

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Abstract

The invention provides a preparation method and application of a hydrogel electrode and relates to hydrogel electrodes. The preparation method of the hydrogel electrode comprises the following steps: with graphite powder as a raw material, adding sodium nitrate, sulphuric acid and potassium hypermanganate, mixing and then carrying out reaction until a thick mixture is formed; then adding pure water for the first time, further carrying out reaction, then adding pure water for the second time and stopping reaction, then adding a hydrogen peroxide solution for removing unreacted potassium hypermanganate, washing, centrifuging, and drying, so that graphite oxide solids are obtained, and carrying out ultrasonic treatment on the graphite oxide solids, so that a uniformly dispersed graphene oxide aqueous solution is obtained; mixing the graphene oxide aqueous solution with DNA of milt, then adding the mixture into a centrifugal tube to be heated, and after gel is stably formed, inserting a copper wire into a small hole in the bottom of the centrifugal tube to be fixed, so that a graphene oxide and milt-DNA compounded hydrogel electrode is obtained. The graphene oxide and milt-DNA compounded hydrogel electrode can be used for preparing a graphene oxide and milt-DNA composite hydrogel biosensor and can be applied to detection of mutation of mitochondrial DNA of ovarian cancer.

Description

technical field [0001] The invention relates to a hydrogel electrode, in particular to a preparation method and application of a hydrogel electrode which can be used for electrochemical detection of ovarian cancer mitochondrial DNA mutation. Background technique [0002] As one of the malignant tumors that seriously threaten women's health, ovarian cancer is hidden and lacks effective screening methods. It is difficult to effectively improve the early diagnosis rate of ovarian cancer with a variety of clinical tests. [1-3] The high mutation rate of mitochondrial DNA and the high copy number in cancer cells make it an effective molecular marker for non-invasive diagnosis of tumors. [4-5] Conventional mutation analysis methods such as ultrasonic diagnosis, computed tomography (CT) and magnetic resonance (MRI) have the disadvantages of high cost and complicated process. In contrast, electrochemical methods are simple, portable, and low-cost. , high sensitivity, fast, and no ne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J3/075C12Q1/68G01N27/00
Inventor 翁建孙莉萍胡楠彭健
Owner XIAMEN UNIV
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