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Preparation method of western blot sensor with modified magnetic electrode by one-step deposition method

A technology for western imprinting and electrode modification, which is applied in the field of electroanalytical chemistry and protein recognition sensors, to achieve high sensitivity, improve sensitivity, improve conductivity and surface area

Active Publication Date: 2020-12-29
MINJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

MIECS has been successfully used to identify various small molecules, however, it is still very challenging to imprint some larger molecular weight molecules, such as biological macromolecules, especially proteins

Method used

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  • Preparation method of western blot sensor with modified magnetic electrode by one-step deposition method
  • Preparation method of western blot sensor with modified magnetic electrode by one-step deposition method
  • Preparation method of western blot sensor with modified magnetic electrode by one-step deposition method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Graphene@Fe 3 o 4 Preparation of nanomaterials: 40 mg graphene powder was dispersed in 40 g ethylene glycol solution, ultrasonically dispersed, and 0.2 g FeCl was added sequentially 3 ·6H 2 O , 1.8 g sodium acetate, 0.8 g PEG 20000, fully dissolved, transferred to a 50 mL reactor, heated at 200 °C for 10 h; the obtained magnetic graphene@Fe 3 o 4 Nanomaterials were separated by a magnetic field, washed several times with ethanol and pure water, and dried at 60°C. The magnetic graphene@Fe 3 o 4 The morphology of nanomaterials is characterized by transmission electron microscopy, such as figure 2 As shown, it can be seen that Fe 3 o 4 Attached to the surface of graphene flakes.

Embodiment 2

[0038] Preparation of electrodes for western blotting

[0039] 0.1 mg of magnetic graphene@Fe prepared in Example 1 3 o 4 Nanomaterials dispersed in containing 0.2wt% HAuCl 4 In the buffer solution with a pH of 9.0, the polished magnetic glassy carbon electrode is used as the working electrode, and a three-electrode system is used, inserted into the solution, and the cyclic voltammetry scan is performed in the range of -1.4 to 0.6 V, and the scan is 6 circles, and the scan rate is 50mV / s. , rinse the modified electrode with ultrapure water;

[0040] Soak the above-mentioned modified electrode in a mixed solution containing 15 mmol / L mercaptophenylboronic acid and cysteine, self-assemble for 30 minutes, take out the electrode, rinse with ultrapure water, let stand, dry, and then soak in 0.5 mg / L mL of bovine hemoglobin solution, self-assembled for 60 min, took out the electrode, rinsed with ultrapure water, stood still, and dried;

[0041] 22.5 mg acrylamide, 3 mg methylene...

Embodiment 3

[0044] Soak the western blotting electrode prepared in Example 2 in BHb solutions of different concentrations, absorb for 40min, take out the electrode, rinse with ultrapure water, let it stand, dry, and place it in a solution containing 0.1 mol / L KCl, 5mmol / L DPV scanning in potassium ferricyanide solution, the scanning range is -0.2 ~ 0.6 V, the change of peak current is monitored, and the DPV curves of different concentrations of BHb adsorbed by the MIP modified electrode are obtained ( Figure 4 ) and peak current curves with different equilibrium concentrations ( Figure 5 ),like Figure 4 as shown, Figure 4 The medium adsorption time is 40 minutes; the concentration increases from top to bottom: 0.0 μg / mL, 0.0001 μg / mL, 0.001 μg / mL, 0.01 μg / mL, 0.1 μg / mL, 0.5 μg / mL, 1.0 μg / mL, 5.0 μg / mL, 10 μg / mL, 50 μg / mL, 100 μg / mL, 500 μg / mL, 1000 μg / mL; with the increase of BHb concentration, the peak current signal gradually weakened, indicating that the imprinted holes on the e...

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Abstract

The invention discloses a fabrication method for a protein electrochemical imprint sensor of a modified magnetic electrode by a one-step deposition method, and relates to the technical field of electrical analysis chemistry and protein recognition sensors. The fabrication method comprises the steps of 1) preparing a graphene@Fe3O4 nanometer composite material; 2) fabricating a graphene@Fe3O4@Au modified magnetic electrode by the one-step electrodeposition method; 3) introducing boric acid, amino and carboxyl active group into a surface of the modified magnetic electrode so that a plurality ofrecognition sites are provided for protein imprint; 4) pre-assembling protein; 5) performing electrochemical polymerization on a protein imprint membrane; and 6) washing template protein with an eluent so as to build the protein molecular imprint electrochemical sensor. The magnetic modified electrode fabricated by the method is beneficial for introducing the active group, increasing the recognition sites, improving selectivity, expanding surface area and improving conductivity of electrode surface, the built protein imprint electrochemical sensor is high in sensitivity and can be used repeatedly, and target protein can be specially recognized.

Description

technical field [0001] The invention relates to a preparation method of a protein electrochemical imprinting sensor modified by a one-step deposition method to modify a magnetic electrode, and relates to the technical fields of electroanalytical chemistry and protein recognition sensors. Background technique [0002] Biorecognition elements, such as enzymes, antibodies, aptamers, etc., have been widely used in the development of biosensors in recent years. However, although these recognition elements have good recognition ability for target analytes, their application is limited due to their instability in the natural environment, harsh use conditions, and cumbersome preparation. To overcome these shortcomings, molecularly imprinted polymers (MIPs), as a new type of biorecognition element, have the characteristics of high stability, low price, reusability, and easy preparation, and have become ideal biomimetic materials. [0003] Molecular imprinting technology, also known ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
CPCG01N27/308G01N27/3277G01N27/3278
Inventor 李艳霞林兴乐赖嘉嘉钱承秀
Owner MINJIANG UNIV
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