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A preparation method of graphene oxide/alpha-fetoprotein aptamer electrochemical sensor

An alpha-fetoprotein and graphene technology, applied in scientific instruments, instruments, material analysis by electromagnetic means, etc., can solve the problems of being susceptible to external interference, loss of labeled antibodies, long analysis time, etc., to overcome poor stability. , the effect of increasing the amount of modification and simplifying the analysis steps

Active Publication Date: 2019-10-01
青岛远诚创智科技有限公司
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  • Claims
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Problems solved by technology

However, these methods have inherent defects, such as the loss of some labeled antibodies, radiation hazards and environmental pollution, etc.; moreover, the lanthanide markers required in fluorescent immunoassays are expensive and susceptible to external interference
In addition, the above analysis methods require a long analysis time and require skilled operators

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  • A preparation method of graphene oxide/alpha-fetoprotein aptamer electrochemical sensor
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  • A preparation method of graphene oxide/alpha-fetoprotein aptamer electrochemical sensor

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[0031] It has been found through experiments that different process parameters have a certain influence on the obtained graphene oxide / alpha-fetoprotein aptamer complex. From the perspective of improving detection sensitivity, the present invention obtains a A preparation method of graphene oxide / alpha-fetoprotein aptamer complex, through which a sufficient amount of aminated alpha-fetoprotein is grafted onto carboxylated graphene oxide to maximize the sensitivity of the electrochemical sensor. The preparation method comprises the following steps:

[0032] (1) ultrasonically disperse 2 to 6 mg of graphene oxide in 1 to 3 ml of deionized water for 1 to 1.5 hours to obtain a uniformly dispersed graphene oxide suspoemulsion;

[0033] (2) adding 0.3~0.6g NaOH and 0.2~0.5g chloroacetic acid to the obtained graphene oxide suspoemulsion, and ultrasonic treatment for 10~12h, to increase the carboxyl groups on the graphene oxide;

[0034] (3) After centrifuging and washing the carboxy...

Embodiment 1

[0057] The preparation method of graphene oxide / alpha-fetoprotein aptamer complex comprises the following steps:

[0058] (1) Ultrasonic disperse 2 mg of graphene oxide in 1 ml of deionized water for 1 h to obtain a uniformly dispersed graphene oxide suspoemulsion.

[0059] (2) Add 0.3g NaOH and 0.2g chloroacetic acid to the above graphene oxide suspoemulsion, and ultrasonically treat it for 12h to increase the carboxyl groups on the graphene oxide. Such as figure 2 As shown, graphene oxide has a large change in its corresponding impedance spectrum before and after carboxylation treatment, indicating that the carboxyl functional group content on the surface of graphene oxide after carboxylation increases.

[0060] (3) The carboxylated graphene oxide was washed by centrifugation, then 1.5ml of 0.3M EDC and 1ml of 0.2M NHS were added, and ultrasonic treatment was performed for 1h to activate the carboxyl groups on the graphene oxide.

[0061] (4) After centrifuging and washin...

Embodiment 2

[0064] The preparation method of graphene oxide / alpha-fetoprotein aptamer complex comprises the following steps:

[0065] (1) Ultrasonic disperse 4 mg of graphene oxide in 1.5 ml of deionized water for 1 hour to obtain a uniformly dispersed graphene oxide suspoemulsion.

[0066] (2) Add 0.4g NaOH and 0.3g chloroacetic acid to the above-mentioned graphene oxide suspoemulsion, and ultrasonically treat it for 12h to increase the carboxyl groups on the graphene oxide.

[0067] (3) The carboxylated graphene oxide was washed by centrifugation, then 3ml of 0.3M EDC and 2ml of 0.2M NHS were added, and ultrasonic treatment was performed for 1h to activate the carboxyl groups on the graphene oxide.

[0068] (4) After centrifuging and washing the activated graphene oxide, add 0.75 ml of aminated alpha-fetoprotein aptamer solution (prepared with Tris-EDTA solution) with a concentration of 200 μg / ml and incubate for 30 min to bind the alpha-fetoprotein aptamer chain grafted onto the graph...

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Abstract

The invention discloses a preparation method of a graphene oxide / alpha fetoprotein aptamer electrochemical sensor. The preparation method comprises the following steps: taking carboxylated graphene oxide as a substrate, grafting an aminated alpha fetoprotein aptamer chain on the substrate through carbodiimide reaction, then modifying the obtained compound on an electrode, closing a non-specific adsorption site on the modified electrode with bovine serum albumin, so that a graphene oxide / alpha fetoprotein aptamer electrochemical sensing interface is formed; and taking the modified electrode as a working electrode, and investigating variation, caused by alpha fetoprotein solution with different concentrations, of an impedance signal on the sensing interface by adopting an impedance method. Compared with an existing alpha fetoprotein detection method, the preparation method has the advantages that an alpha fetoprotein aptamer is adopted for analyzing a target protein, harms such as redundant multiple labeling operation, radiation and pollution do not exist, and the sensor has the advantages of high sensitivity, good selectivity, simplicity in preparation, high analysis speed, mild reaction conditions, low preparation cost and the like.

Description

technical field [0001] The invention belongs to the technical field of novel functionalized nanomaterials and electrochemical sensing, and in particular relates to a preparation method of a graphene oxide / alpha-fetoprotein aptamer electrochemical sensor. Background technique [0002] Alpha-fetoprotein (AFP), a serum glycoprotein, is currently the most important biomarker for the diagnosis of primary liver cancer (Hepatocellular Carcinoma, HCC). In addition, abnormal levels of alpha-fetoprotein in serum are also related to some other related diseases, such as neural tube defects, brain / spinal cord malformations, and chromosomal abnormalities. Therefore, the detection of serum alpha-fetoprotein content is of great significance to the clinical diagnosis of liver cancer and the early screening of some diseases. [0003] At present, the determination of alpha-fetoprotein content in serum is mainly carried out by immunoassay in clinical detection. Conventional immunoassays inclu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N27/327
Inventor 张菲菲王宗花杨少红管嵩
Owner 青岛远诚创智科技有限公司
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