Sensor preparation method based on ECL-RET action between GO and GQDs and application on kinas detection

A sensor and action technology, applied in chemical instruments and methods, analysis by chemical reaction of materials, inorganic chemistry, etc., can solve the problem of less research and application of GQDs, and achieve high sensitivity, good stability, and low detection limit. Effect

Inactive Publication Date: 2014-01-15
NANCHANG UNIV
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  • Sensor preparation method based on ECL-RET action between GO and GQDs and application on kinas detection
  • Sensor preparation method based on ECL-RET action between GO and GQDs and application on kinas detection
  • Sensor preparation method based on ECL-RET action between GO and GQDs and application on kinas detection

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Embodiment 1

[0021] (1) GO was prepared by the Hummers method: 1.0 g graphite and 1.0 g NaNO 3 Added to 46 mL of 98% H 2 SO 4 , add 6.0 g KMnO slowly under ice bath 4 , stirred in a water bath at 35 °C for 1 h, added 80 mL of ultrapure water, continued to stir for 30 min, and then added 200 mL of ultrapure water, then added 6 mL of 30% H 2 o 2 , react at room temperature for 1 h; filter the product while it is hot and wash it with ultrapure water until the filtrate is neutral, disperse the product in 500 mL ultrapure water, and sonicate for 2 h to obtain uniformly dispersed GO;

[0022] (2) Preparation of GQDs by hydrothermal method: the dried GO was placed in a tube furnace under N 2Under the condition of protection, be heated to 200 ° C with the heating rate of 5 ° C / min and keep 2 h, obtain graphene sheet; 0.05 g graphene sheet is added to the concentrated sulfuric acid that volume ratio is 1:3:Concentrated nitric acid Sonicate in the mixed solution for 17 h, in which the mass perc...

Embodiment 2

[0029] Preparation process of ECL biosensor

[0030] (1) Pretreatment of the glassy carbon electrode: Before the glassy carbon electrode is modified, the α-Al 2 o 3 Polish in the paste, and then ultrasonically clean with ethanol and water for 1 min;

[0031] (2) The preparation process of ECL biosensor is as follows: figure 1 shown. 10 μL of chitosan solution with a mass percent concentration of 0.5% was drop-coated on the surface of the glassy carbon electrode and dried, then the electrode was immersed in the GQDs-COOH solution containing 5 mM EDC, and incubated at room temperature for 5 h; After washing with 10 mM, pH 7.4 PBS buffer solution, insert the electrode into a solution containing 5 mM EDC, 8 mM NHS and 50 μM peptide for 3 h; then insert the electrode into Tris buffer containing CK2 and ATP React in the solution for 2 h to phosphorylate the polypeptide; incubate the phosphorylated polypeptide modified electrode in the Ab-GO solution for 1 h to prepare the ECL se...

Embodiment 3

[0036] ECL biosensor for detection of CK2

[0037] Under optimal experimental conditions, the ECL biosensor constructed using the ECL-RET interaction between GO and GQDs detected CK2 activity. Depend on Figure 6 A shows that with the increase of CK2 concentration, the ECL signal of GQDs gradually decreased, and when the CK2 concentration was 30 U / mL, the ECL intensity reached the maximum. Figure 6 B is the standard curve for CK2 detection. When the CK2 concentration is 0.05-5 U / mL, the CK2 concentration has a good linear relationship with the ECL signal. The linear equation is I = 1621.6 - 142.7c (I is the ECL intensity, c is the CK2 concentration) , and the detection limit was 0.023 U / mL. The detection limit of the method is lower than that of the electrochemical method and the fluorescence method, and the linear range is wider, indicating that the ECL sensor proposed by the invention can detect kinase activity with high sensitivity.

[0038] in the presence of 0.1 M Na ...

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Abstract

The invention discloses a sensor preparation method based on an ECL-RET action between GO and GQDs and an application on kinas detection, and belongs to the field of electrochemiluminescence (ECL). The preparation method comprises the following steps: coating chitosan on the surface of an electrode, and orderly assembling graphene quantum dots and polypeptides onto the surface of the electrode through a covalent interaction. Under the actions of protein kinase and triphosadenine, the polypeptides carry out phosphorylation reactions, through the specific recognition action between an antibody and an antigen, oxidized graphene conjugated with a phosphorylated antibody is assembled to the phosphorylated serine sites of the polypeptide, thus the distance between the oxidized graphene and the graphene quantum dots is narrowed down, so that the electrochemiluminescence (ECL) of graphene quantum dots is quenched. The larger the concentration of protein kinase is, the more phsophorylated sites are generated on the polypeptide modified electrode surface, the more oxidized graphene is assembled on a sensing interface, the stronger the electrochemiluminescence quenching effect of graphene quantum dots will be, and thus the high sensitive detection on protein kinase is achieved.

Description

technical field [0001] The invention relates to the field of electrochemiluminescence, in particular to a sensor preparation method and kinase detection application based on the ECL-RET interaction between GO and GQDs. Background technique [0002] Graphene is a two-dimensional free-state atomic crystal, which is used to construct sp 2 The basic structural unit of hybrid carbon has attracted great attention from researchers at home and abroad since it was first reported in 2004. The excellent electrochemical, mechanical and thermodynamic properties of graphene make it widely used in the research of polymer materials, sensors, energy-related materials and field effect transistors. However, graphene is a zero-bandgap material, and it is difficult to observe its luminescence phenomenon. Therefore, using various methods to deal with the band gap of graphene and expand its light-emitting applications has aroused great research interest of scientists, such as opening the band ga...

Claims

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Application Information

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IPC IPC(8): G01N21/76C01B31/04C01B32/184C01B32/194
Inventor 邱建丁向彩云梁汝萍
Owner NANCHANG UNIV
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