An electrochemical immunosensor for detecting Alzheimer's disease markers, its preparation method and application

A technology of Alzheimer's disease and immunosensor, which is applied in the field of electrochemical immunoassay and biosensing, can solve the problems of cumbersome detection methods, high detection limit, and complicated operation, so as to improve the sensitivity and detection limit and avoid detection errors , the effect of increasing the comparison area

Active Publication Date: 2021-06-15
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 detection methods are cumbersome, complicated to operate, expensive or have high detection limits. A fast, simple and sensitive detection method is of great significance

Method used

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  • An electrochemical immunosensor for detecting Alzheimer's disease markers, its preparation method and application
  • An electrochemical immunosensor for detecting Alzheimer's disease markers, its preparation method and application
  • An electrochemical immunosensor for detecting Alzheimer's disease markers, its preparation method and application

Examples

Experimental program
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Effect test

Embodiment 1

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

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

[0046] First, the gold nanobicones were prepared by the seed growth method, and the specific steps were as follows:

[0047] Preparation of seed solution:

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

[0049] Growth solution preparation: Add 2 mL of HAuCl sequentially to 40 mL of CTAB (0.1 M) solution 4 (0.01M), 0.4mL AgNO 3 (0.01M), 0.8mL HCl (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 s, and leave overnight at room temperature;

[0051] After centrifugation, the precipitate was dispersed in 10 mL of CTAB solution (0.01M) to obtain gold nanobipyramids (AuB...

Embodiment 2

[0066] 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 (disease markers in step (5) in Example 1 Apolipoprotein E4 solution concentration is 1ng / mL), tested in 20 mL, 10 mM phosphate buffer solution 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 600s;

[0069] (3) When the background current was stable, 20 μL of 1M hydrogen peroxide solution was dropped into 20 mL of 10 mM phosphate buffer solution with a pH of 7.4 every 50 s, and the current change was recorded.

Embodiment 3

[0071] Performance Analysis of Prepared Sensors

[0072] In order to evaluate the performance of the prepared electrochemical immunosensor, ApoE4 standards with different concentrations (0.1, 1, 10, 100, 1000, 10000, 20000 ng / mL) prepared in phosphate buffer with pH 7.4 were prepared Different immunosensors were analyzed. Test the current change values ​​corresponding to different concentrations of standard substances by time current method, and draw a 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 detection of 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 present invention relates to an electrochemical immunosensor for detecting Alzheimer's disease markers, its preparation method and application, specifically comprising the following steps: ① pretreatment of platinum carbon electrodes, and graphene oxide quantum The dots are deposited on the platinum carbon electrode; ②The prepared gold bicone-coated platinum nanostructure is used as the substrate material, and it is deposited on the surface of the graphene oxide quantum dots by electrophoretic deposition, so as to further improve the Alzheimer's disease marker capture antibody and accelerate the electron transfer rate on the electrode surface; ③The prepared AuPd‑PDA nanocomposite material is used as a signal amplification label to immobilize the detection antibody; through the synergistic effect of the substrate material and the AuPd‑PDA composite nanomaterial, the sensor’s sensitivity is further improved Sensitivity; the prepared electrochemical immunosensor can be used for the detection of Alzheimer's disease markers; it has the characteristics of strong specificity, high sensitivity and low 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, a preparation method and application thereof. Background technique [0002] Alzheimer's disease (AD) is a neurodegenerative disease characterized by cognitive impairment and behavioral impairment, and is the most common form of dementia, accounting for about 50-60% of dementia cases. %. The clinical manifestations are memory loss, language barrier, mental retardation, slow thinking and other symptoms, and even death in severe cases. AD is a very complex disease, often without obvious genetic features, for which age appears to be the main risk factor. The complexity of AD diagnosis determines that it has a very broad clinical definition, and some external environmental factors and internal genetic factors will further aggravate the condition. External factors mainly include...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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