Electrochemical immunosensor for detecting Alzheimer's disease markers and preparation method and application thereof

An Alzheimer's disease, immunosensor technology, applied in the field of electrochemical immunoassay and biosensing, can solve the problems of cumbersome detection methods, high cost, high detection limit, improve sensitivity and detection limit, avoid detection errors, The effect of increasing the comparison area

Active Publication Date: 2019-07-12
郑州森旺智慧餐饮管理有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are many detection methods for Alzheimer's disease markers, such as radioimmunoassay, electrochemiluminescence, colorimetry, etc., but most of the de

Method used

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  • Electrochemical immunosensor for detecting Alzheimer's disease markers and preparation method and application thereof
  • Electrochemical immunosensor for detecting Alzheimer's disease markers and preparation method and application thereof
  • Electrochemical immunosensor for detecting Alzheimer's disease markers and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] A preparation method of an electrochemical immunosensor for detecting Alzheimer's disease markers, the steps are as follows:

[0045] 1. Preparation of gold bicone-coated platinum nanostructures

[0046] First, prepare gold nano-bicones by seed growth method, the specific steps are as follows:

[0047] Preparation of seed solution:

[0048] Add 250μL of sodium citrate (0.01M) and 125μL of HAuCl to 9.625mL of ultrapure water in sequence 4 (0.01M), 150μLNaBH 4 (0.01M) Stir well, put it at room temperature for two hours before use;

[0049] Growth solution preparation: Add 2mL HAuCl to 40mL CTAB (0.1M) solution sequentially 4 (0.01M), 0.4mLAgNO 3 (0.01M), 0.8mLHCl (1M), 0.32mL ascorbic acid (0.1M);

[0050] Add 0.5 mL of seed solution to 9.5 mL of growth solution, stir for 30 seconds, overnight at room temperature;

[0051] Centrifugal separation, disperse the precipitate in 10 mL CTAB solution (0.01M) to obtain gold nano double cones (AuBP), such as figure 1 As shown, the obtained ...

Embodiment 2

[0066] For the detection of Alzheimer's disease marker ApoE4, the steps are as follows:

[0067] (1) Use an electrochemical workstation to test with a three-electrode system, the Ag / AgCl electrode is the reference electrode, the platinum wire electrode is the auxiliary electrode, and the prepared sensor is the working electrode (the disease marker in step (5) in Example 1 The concentration of apolipoprotein E4 solution is 1ng / mL), tested in 20 mL, 10 mM phosphate buffer with pH 7.4;

[0068] (2) Use the time-current method to detect the current response value of hydrogen peroxide, the input voltage is -0.2V, the sampling interval is 0.1s, and the running time is 600 s;

[0069] (3) When the background current is stable, add 20 μL of 1M hydrogen peroxide solution to 20 mL of 10 mM phosphate buffer with pH 7.4 every 50s, and record the current change.

Embodiment 3

[0071] Performance analysis of prepared sensors

[0072] In order to evaluate the performance of the prepared electrochemical immunosensor, different concentrations of ApoE4 standards (concentrations of 0.1, 1, 10, 100, 1000, 10000, 20000 ng / mL) prepared in phosphate buffer with pH 7.4 were prepared Different immunosensors for analysis. Use the time current method to test the current change value corresponding to different concentration standards, and draw the standard curve, (such as Figure 4 As shown, ΔI=18.191LgC+200.78; R 2 =0.9841) such as Figure 4 Shown. The detection limit of the sensor of the present invention for the Alzheimer's disease marker ApoE4 is 0.03 ng / mL (S / N sex to noise ratio is 3), and the linear range is 0.1 ng / mL-2 μg / mL.

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Abstract

The invention relates to an electrochemical immunosensor for detecting Alzheimer's disease markers and a preparation method and application thereof. The preparation method specifically comprises the following steps of 1, pretreating a platinum carbon electrode, and depositing graphene oxide quantum dots on the platinum carbon electrode through an electric deposition manner; 2, taking a prepared gold biconical platinum coated nanostructure as a base material, and depositing the gold biconical platinum coated nanostructure on the surfaces of the graphene oxide quantum dots through an electrophoretic deposition method so as to further improve the fixing amount of an Alzheimer's disease marker capturing antibody and improve the electron transfer rate on the electrode surface; and 3, taking a prepared AuPd-PDA nanocomposite as a signal amplification tag to fix a detection antibody. Through the synergistic action of the base material and the AuPd-PDA nanocomposite, the sensitivity of the sensor is further improved; the prepared electrochemical immunosensor can be used for detecting Alzheimer's disease markers; and the electrochemical immunosensor has the characteristics of being strong in specificity, high in sensitivity and low in detection limit.

Description

Technical field [0001] The invention belongs to the technical field of electrochemical immunoassay and biosensing, and provides an electrochemical immunosensor for detecting Alzheimer's disease markers, and a preparation method and application thereof. Background technique [0002] Alzheimer's disease (AD) is a neurodegenerative disease characterized by cognitive impairment and behavioral impairment. It is the most common form of Alzheimer’s disease, accounting for about 50-60% of Alzheimer’s disease. %. The clinical manifestations are symptoms such as memory loss, language impairment, mental retardation, and slow thinking, which can even lead to death in severe cases. AD is a very complex disease, usually without obvious genetic characteristics, it seems that age is its main risk factor. The complexity of diagnosis of AD determines that it has a broad clinical definition. Some external environmental factors and internal genetic factors will further aggravate the disease. The ...

Claims

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Application Information

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IPC IPC(8): G01N27/327
CPCG01N27/3278
Inventor 刘一镳何广莉牛鹏飞侯梦丹秦伟鹏张平平张守仁杨保成
Owner 郑州森旺智慧餐饮管理有限公司
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